Creative Genius and Psychopathology

Creative Genius and Psychopathology
Creative Genius and Psychopathology

Creative Genius and Psychopathology Is there a Link Between Creative Genius and Psychopathology?

Do you think that society places too much or too little emphasis on creative achievement and why?

Is Parenthood Associated with Happiness?

  1. How might happiness influence whether a person decides to have children?
  2. Why do you think the association between life satisfaction and children was not found in other countries?

What Motivates Suicide Bombers?

  1. How can we understand the motivational dynamics that separate those who embrace violent versus nonviolent tactics?
  2. How do suicide bomber’s motives compare to those of a soldier who dies for a cause?

Is There One Really Great Personality?

  1. What do you think are the defining features of a great personality?
  2. How might the cultural context influence whether a person’s personality is judged as optimal?

Why Does a Cell Phone Look Like a Gun?

  1. How might this research be used to prevent future tragic mistakes?
  2. Why would Black police officers be just as likely as White officers to mistake a tool for a gun when primed with a Black face?

Does Everyone Have ADHD?

  1. Would ADHD be as controversial if the treatment did not involve drugs? Why or why not?
  2. Do you think ADHD would be diagnosed as often as it is if drugs were not readily available for its treatment?

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Bipolar 2 disorder Case Study Assignment

Bipolar 2 disorder
Bipolar 2 disorder

Bipolar 2 disorder

A case study on a 16 year-old girl with ç

  1. Identify the target symptoms for a child or adolescent from your practicum site. Must be a different case then the first one.

2 State the DSM-5 diagnosis with evidence.

  1. Identify two differential diagnosis with rationale.
  2. Interventions

List non-pharmacological interventions with rationale.

Psycho-pharmacological intervention with rationale.

Include a sample prescription

Provide a link to the medication education sheet to be given to the family/patient.

Provide a link to the appropriate diagnostic sheet to be given to the family/patient.

  1. List additional assessments required ie screening tools, labs with rationale.
  2. Name a community resource / referrals for the patient/family. (It can be a web site for a support group; literature, organizations, educational support, or a list of community resources etc.)
  3. Three scholarly references in APA format

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Korean Students and Mental Health to Parents

Korean Students and Mental Health to Parents Effects of Strategy Type to Disclose Korean Students and Mental Health to Parents

Korean Students and Mental Health to Parents
Korean Students and Mental Health to Parents

Research Question: (please rewrite in a more advanced way if possible)

Imagine you are a student with mental health issues. Your psychiatrist has deemed that you have depression, bipolar disorder, or any other type of disorder and may need to take medication. How would you tell your parents? Would you tell them at all?

Participants: 15 born and raised Korean students will be given a real-life situation in which they must Need to reference:

Formula: Wx = D(S, H) + P(H, S) + Rx

Wx is the weight which refers to the weightiness of the act.

D(S, H) symbolizes the distance between the distance and speaker

P(H, S) + R stands for the power o the hearer over the speaker

Rx implies the degree of the imposition of the act

Needs to include:

A section on Korean culture regarding mental health Quantitative Measurement

Quantitative Molecular Imaging of Neurotransmitters

Quantitative Molecular Imaging of Neurotransmitters Direct Targeted Quantitative Molecular Imaging of Neurotransmitters in Brain Tissue Sections

Quantitative Molecular Imaging of Neurotransmitters
Quantitative Molecular Imaging of Neurotransmitters

Mohammadreza Shariatgorji,1 Anna Nilsson,1 Richard J.A. Goodwin,1,2 Patrik Ka® back,1 Nicoletta Schintu,3

Xiaoqun Zhang,3 Alan R. Crossman,4 Erwan Bezard,5 Per Svenningsson,3 and Per E. Andren1,

* 1Biomolecular Imaging and Proteomics, National Center for Mass Spectrometry Imaging, Department of Pharmaceutical Biosciences,

Uppsala University, P.O. Box 591 BMC, 75124 Uppsala, Sweden

2AstraZeneca R&D, Alderley Park, Macclesfield, Cheshire SK10 4TF, UK

3Center for Molecular Medicine, Department of Neurology and Clinical Neuroscience, Karolinska Institutet and Karolinska University Hospital, 17176 Stockholm, Sweden

4Faculty of Life Sciences, University of Manchester, Manchester M13 9PL, UK

5Universite¥ de Bordeaux, Institut des Maladies Neurode¥ ge¥ ne¥ratives, UMR 5293 Bordeaux, France

*Correspondence: per.andren@farmbio.uu.se

http://dx.doi.org/10.1016/j.neuron.2014.10.011

SUMMARY

Current neuroimaging techniques have very limited abilities to directly identify and quantify neurotransmitters from brain sections. We have developed a molecular-specific approach for the simultaneous imaging and quantitation of multiple neurotransmitters, precursors, and metabolites, such as tyrosine, tryptamine, tyramine, phenethylamine, dopamine, 3-methoxytyramine, serotonin, GABA, glutamate,

acetylcholine, and L-alpha-glycerylphosphorylcholine, in histological tissue sections at high spatial resolutions. The method is employed to directly measure changes in the absolute and relative levels of neurotransmitters in specific brain structures in animal disease models and in response to drug treatments, demonstrating the power of mass spectrometry imaging in neuroscience.

Quantitative Molecular Imaging of Neurotransmitters Introduction

Small-molecule neurotransmitters such as the catecholamine dopamine (DA), the amino acids g-aminobutyric acid (GABA) and glutamate (Glu), and acetylcholine (ACh) are critical chemical messengers that transmit signals between neurons. Changes in their concentrations are associated with numerous normal neuronal processes, such as sleep and aging, but also several disease states, including Alzheimerís disease, Parkinsonís disease (PD), depression, drug addiction, and attention deficit hyperactivity disorder. A better understanding of their relative abundance and distribution would provide insights into these complex neurological processes and disorders. At present, researchers rely on indirect histochemical, immunohistochemical (IHC), and ligand-based assays to detect these small-molecule transmitter substances (de Jong et al., 2005; Falck et al., 1962;

Jones and Beaudet, 1987). The antibodies used in IHC often have major limitations, such as an inability to distinguish between different transmitters (Keenan and Koopowitz, 1981).

Nuclear medicine imaging is a widely used tool for indirect visualization of the distribution, abundance, and activity of neurotransmitters in the brain, relying on radiolabeled tracers

that emit gamma rays, which are detected using positron emission

tomography (PET) or single photon emission computed tomography scanners (Pimlott and Sutherland, 2011). Developing

appropriate labeled compounds is often complicated,

particularly for studying endogenous chemical messengers

(Badgaiyan, 2011). There is often an inability to discriminate

between the labeled parent compound and a metabolite retaining

the label. Imaging techniques that can directly visualize the

localization of neurotransmitters and simultaneously quantitate

them without relying on a label would thus represent a scientific

breakthrough. Another significant advance in the molecular

analysis of neurological tissues would be the development of

a multiplex method that enables the simultaneous detection of

multiple endogenous compounds and pharmaceutical agents.

MALDI mass spectrometry (MS) imaging has been used

for direct molecule-specific compound detection, distribution

mapping, and identifying molecular species in tissue sections

(Caprioli et al., 1997; Cornett et al., 2007). It can generate multiplex

full mass spectrum data at near-cellular spatial resolution.

Crucially, it provides a powerful tool for in situ visualization of

target compounds and for determining their abundance and

spatial distribution without requiring a great deal of a priori information.

Moreover, the data gathered during such experiments

can be used to simultaneously identify unknown endogenous

molecular changes due to disease or degeneration. MALDI-MS

can be used for the direct analysis of targets, ranging from small

molecules to large proteins (Schwamborn and Caprioli, 2010).

However, despite some notable successes, it is relatively insensitive

toward certain compound classes and sometimes suffers

from signal interference caused by matrix clusters/fragments

(Sugiura et al., 2012; Ye et al., 2013). To date, it has found only

limited applications in the study of neurotransmitters due to

their poor ionization efficiencies. As such, the development of

a reagent that would enable selective neurotransmitter derivatization

while also facilitating their detection by MS and acting

as an efficient matrix to assist their laser-induced desorption

and ionization would be a significant breakthrough.

Neuron 84, 697ñ707, November 19, 2014 ™2014 Elsevier Inc. 697

Here we describe a unique approach for the direct and

absolute quantitative imaging of classical and common neurotransmitters

in histological brain tissue sections. We report

an example of the simultaneous mapping of biogenic amines

and amino acid chemical messengers following in situ derivatization

in brain tissue sections. The method is employed to

simultaneously measure changes in the levels of neurotransmitters,

including tyrosine, tryptamine, tyramine, phenethylamine,

DA, 3-methoxytyramine (3-MT), serotonin (5-HT), GABA, and

Glu, in specific brain structures of primates and rodents with

and without DA depletion and L-3,4-dihydroxyphenylalanine

(L-DOPA) treatment. In addition, we have developed a method

that can be used to image and quantitate ACh and an ACh

precursor, L-alpha-glycerylphosphoryl-choline (alpha-GPC) in

brain sections, as well as to study activity within cholinergic

pathways by measuring changes in ACh concentrations, imaged

at a 15 mm spatial resolution. Together, these experiments

demonstrate a significant advancement in the molecular imaging

arsenal available to neuroscientists.

RESULTS AND DISCUSSION

Without modification, most neurotransmitters are not directly

detectable by MALDI-MS due to their poor ionization efficiency

and the overlapping signals of isobaric compounds. To address

the limited ionization and desorption of neurotransmitters, we

developed a method for their in situ chemical derivatization.

Once modified, the neurotransmitters are readily ionized and detected,

enabling their identification and the quantitative mapping

of their distributions. Our strategy uses pyrylium salts such

as 2,4-diphenyl-pyranylium tetrafluoroborate (DPP-TFB), which

react selectively with primary amines to produce N-alkyl- or

N-aryl-pyridinium derivatives (Figure S1, available online). The

reaction proceeds under very mild conditions and occurs rapidly

at ambient temperature and pressure without any need for

stirring or agitation. Such conditions are required to preserve

the localization of endogenous compounds. The resulting

charged derivatives have laser desorption and ionization

efficiencies sufficient to enable the detection of endogenous

small-molecule primary amines (Johannesen et al., 2012; Quirke

et al., 1994), such as tyrosine, tryptamine, tyramine, phenethylamine,

DA, 3-MT, 5-HT, GABA, and Glu, simultaneously in a single

experiment. Importantly, the selective derivatization of the

primary amines separates their signals from those of overlapping

isobaric compounds. A MALDI-MS analysis of a representative

pyrylium derivatization reaction clearly showed that all of the

desired pyridinium ions were formed in high yields (Figure S1).

Furthermore, the DPP-TFB derivatives of endogenous primary

amines, including neurotransmitters and their metabolites and

precursors, undergo self-assisted laser desorption ionization.

As such, they are amenable to MALDI-MS imaging without

needing to be assisted by a matrix such as a-cyano-4-hydroxycinnamic

acid (CHCA).

The combination of pyrylium derivatization and multiplex data

acquisition generates hundreds of ion signals in each MALDI-MS

imaging experiment. This makes it possible to obtain images

showing the distribution of every individual ion within an m/z

window of 200 to 450 in the sample being studied (Figure 1A).

In addition to showing the distributions of known derivatives,

these images can be used to characterize unidentified ions

whose concentrations and localizations may be affected by

neurological events and diseases. We demonstrated this by

determining the localization of selected neurotransmitters in

brain tissue samples derivatized using DPP-TFB and also

2,4,6-trimethyl-pyranylium tetrafluoroborate (TMP-TFB) as a

confirmatory reagent. The identities of the signals for the neurotransmitter

derivatives were verified by performing tandem

MS (MS/MS) experiments using derivatized synthetic standards

alongside the endogenous neurotransmitter. In all cases, the

product ions observed for the standards were identical to those

for the endogenous species. For example, both exogenous and

standard GABA generated a peak at m/z 208.1 and similar MS/

MS product ions when derivatized with TMP-TFB and m/z

318.1 and similar MS/MS product ions when derivatized with DPP-TFB (Figure S1). Similarly, both endogenous and standard DA yielded peaks at m/z 136 and m/z 232, respectively, after derivatization with these reagents (Figure S1). In addition, the elemental compositions obtained for the target ions from accurate mass Fourier transform ion cyclotron resonance (FT-ICR)- MS measurements were in good agreement with the expected structures of the derivatized neurotransmitters (Tables S1 and S2). Deuterated neurotransmitters were used as calibration standards to determine the absolute concentrations of the endogenous neurotransmitters directly in brain tissue sections.

MALDI-MS Imaging of Neurotransmitters in Control Rodent Brain Sections

The in situ neurotransmitter derivatization and imaging procedure was initially demonstrated using coronal and sagittal mouse and rat brain sections (Figures 1, S1, and S2). The distribution

of the pyrylium salt formed by derivatizing GABA with DPPTFB

(m/z 318.1) in coronal rat brain tissue sections showed

that GABA is most abundant in the medial septum/diagonal

band region (MSDB) of the brain (Figure S2), which is consistent

with previous reports (Alreja et al., 2000). The MSDB provides a

major GABAergic input to the hippocampus (Alreja et al., 2000;

Qin and Luo, 2009). Imaging of sagittal rat brain sections (Figure

1B) revealed structures with high GABA concentrations,

including the hypothalamus, midbrain, and basal forebrain (Figure

1C). GABA is a dominant neurotransmitter in the hypothalamus

and plays an important role in hypothalamic inhibitory

circuits (Decavel and Van den Pol, 1990). Moreover, the basal

forebrain and its substructures are reported to provide major

GABAergic inputs to the hippocampus (Alreja et al., 2000). The

importance of midbrain neurons in GABA recycling and the

corelease of GABA from the dopaminergic midbrain neurons

via plasma membrane uptake (Tritsch et al., 2014) may explain

the high GABA concentration in this region. Deuterated GABA

(D6-GABA) was used to determine absolute GABA concentrations

in rat brain tissue sections. The absolute concentration of

GABA in the coronal brain section as a whole was 5 nmol/mg,

while that in the MSDB region was 10 nmol/mg (Figure S2). These

values are consistent with previous results (Schaaf et al., 1985).

Other primary amine neurotransmitters, such as tyrosine,

tryptamine, tyramine, phenethylamine, DA, 3-MT, 5-HT, GABA,

and Glu, were detected and imaged in the same brain tissue

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Quantitative Molecular Imaging of Neurotransmitters

698 Neuron 84, 697ñ707, November 19, 2014 ™2014 Elsevier Inc.

sections that were treated with DPP-TFB to produce 2,4-diphenylpyridinium

derivatives. As before, this substantially increased

the neurotransmitterís MALDI desorption/ionization efficiency

and generated a detectable signal at m/z 362.1 (Figure S1).

Glu was found in most structures of the brain but was mainly

localized in the striatal and cortex regions (Figure S2), which is

consistent with previous reports on the distribution of Glu transporters

(Bressan and Pilowsky, 2000; El Mestikawy et al., 2011;

Herzog et al., 2004). The identity of the Glu peak was verified by

MS/MS fragmentation of derivatized synthetic standards

(deuterated and nondeuterated) and the endogenous signal

(Figure S1).

Analyses of rat brain sagittal sections revealed high DA concentrations

in the substantia nigra, striatum, and ventral pallidum

structures due to the nigrostriatal and mesolimbic DAergic pathways

(Figure 1D). 3-MT, a methylated DA metabolite formed

by the action of the enzyme catechol-O-methyl transferase,

was also imaged and found to have the same distribution as

DA (Figure 1E). 3-MT was initially reported to be an inactive DA

metabolite, but recent studies suggest that it is a neuromodulator

that acts as an agonist of the trace amine-associated receptor

1 (Sotnikova et al., 2010). It is worth noting that distinguishing

between an endogenous compound and its metabolites is

very difficult when using techniques based on radiolabeled

compounds. In the coronal sections (Figure 1F), DA was mostly

localized in the striatal regions (Figure 1G), but it was also

present at lower concentrations in the cingulate cortex, piriform

cortex, medial septal nucleus, and the nucleus of the vertical

limb of the diagonal band (Figure 1H). The intensities of the DA

signals for selected regions (Figure 1F) were used to quantify

their DA levels in relative terms (Figure 1I), revealing that the

average DA concentration in the striatal region is about 40 times

higher than that in the cingulate cortex region.

Neurotransmitter MALDI-MS Imaging of Parkinsonís

Disease Models

To further demonstrate the potential of the derivatization/MALDIMS

imaging methodology, it was used to analyze the distribution

Figure 1. MALDI-MS Imaging Data Displaying

GABA, DA, and 3-MT and Distributions

in Rat Brain Tissue Sections

(AñI) The mass spectrum generated in a MALDIMS

imaging experiment (A) contains hundreds of

ion signals associated with each individual pixel.

These can be used to create images showing the

distributions of each individual ion in the spectrum.

A sagittal tissue section from a rat brain (B) and the

associated relative abundances and distributions

of GABA (C), DA (D), and 3-MT (E). GABA was

found in all structures of the brain but was most

abundant in HY, BF and its substructures, and MB.

High DA concentrations were observed in SN,

CP, VS, and AON. The distribution of 3-MT

mirrored that of DA. DA imaging in a rat brain

coronal tissue section (F) revealed that it was most

abundant in the striatal region (G). By rescaling the

signal intensity of the latter image, it was possible

to detect DA in other parts of the brain, notably

in the A24, Pir, MS, and VDB. However, the

DA concentrations in these substructures were all

much lower than that in striatal region (H). The

intensities of the DA signals in each region of the

brain were compared to enable its relative quantitation.

The inset bars show the rescaled DA signal

intensities for the Pir, MS and VDB, and A24

regions (I). MS data were acquired using either

a MALDI-TOF/TOF (AñE) or a MALDI-FT-ICR

mass spectrometer (GñI). Data are shown using a

rainbow scale, normalized against the total ion

count. Scale bar, 2 and 5 mm. Images were acquired

at spatial resolutions of 100 mm for sagittal

sections and 150 mm for coronal sections. Substantia

nigra, SN; caudate-putamen, CP; ventral

striatum, STRv; anterior olfactory nucleus, AON;

cingulate cortex (area 24), A24; piriform cortex, Pir;

medial septal nucleus, MS; nucleus of the vertical

limb of the diagonal band, VDB; accumbens

nucleus, ACB; hypothalamus, HY; thalamus, TH;

hippocampus, HIPP; corpus callosum, cc; cerebral

cortex, CTX; midbrain, MB; basal forebrain,

  1. See also Figure S1.

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Quantitative Molecular Imaging of Neurotransmitters

Neuron 84, 697ñ707, November 19, 2014 ™2014 Elsevier Inc. 699

of neurotransmitters in experimental models of PD. When studying

neurodegeneration processes in PD models, it is desirable to

assess the integrity and function of the nigrostriatal DAergic

pathway. Various techniques have been used for this purpose,

including the quantitation of dopaminergic neurons and terminals

in the substantia nigra pars compacta and striatum by

immunostaining using an antibody against tyrosine hydroxylase

(TH), and the measurement of TH or DA transporters (DAT) in

striatal tissue extracts by immunoblotting (Ciliax et al., 1995;

Huot et al., 2007). Alternatively, the dopamine neurons can be

quantified by using autoradiography to measure the levels of

a radiolabeled DAT ligand (Boja et al., 1991). The best way to

quantitate DA is to use high-performance liquid chromatography

coupled with an electrochemical detector. However, this

approach provides no information on the neurotransmitterís

anatomical localization. Dopaminergic terminals are heterogeneously

distributed in the striatum; in PD, it is mainly the

dorsolateral regions that exhibit reduced DA levels. Important

information might therefore be lost if the anatomical distribution

of DA is not determined. Our method allows the simultaneous

direct imaging and quantitation of primary amine neurotransmitters

in tissue sections from animal models of PD.

The PD models used in our experiments were generated

by injecting the neurotoxin 6-hydroxydopamine (6-OHDA) into

rodentsí medial forebrain bundles (Zhang et al., 2008) or by

intravenous administration of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine

(MPTP) to primates. Pyrylium derivatization

and MALDI-MS imaging were then used to quantify neurotransmitter

levels in rat brain sections by depositing known concentrations

of deuterated standards onto vehicle control tissue

sections. The DA concentrations in the intact and 6-OHDAlesioned

sides of rat striata were determined to be 124 pmol/mg

Figure 2. MALDI-MS Images of Neurotransmitters

in Coronal Rat PD Model Tissue

Sections

(AñU) Imaging experiments were conducted

on brain tissue sections from unilateral shamlesioned-,

unilateral 6-OHDA-lesioned, and unilateral

6-OHDA-lesioned animals that were treated

with subchronic L-DOPA for 4 weeks, with the

final dose being given as deuterated (D3)-L-DOPA.

The images show the distributions of DA (AñF)

in sham-lesioned (A), 6-OHDA-lesioned (B),

and 6-OHDA-lesioned L-DOPA-treated animals.

Rescaling images (DñF) (with new max intensity

corresponding to 5% of the previous) made it

possible to determine the distribution of DA in

the 6-OHDA-lesioned side of the brain and in

structures with low DA concentrations, such as the

cortex. (D) Rescaled image from (A). (E) Rescaled

image from (B). (F) Rescaled image from (C).

Distribution of D3-DA (formed by the in vivo

breakdown of D3-L-DOPA is imaged in shamlesioned

(G), 6-OHDA-lesioned (H), and 6-OHDAlesioned

L-DOPA-treated animals (I). Distribution

of endogenous 3-MT is imaged in sham-lesioned

(J), 6-OHDA-lesioned (K), and 6-OHDA-lesioned

L-DOPA-treated animals (L), and distribution of

D3-3-MT (derived from D3-L-DOPA) is imaged

in sham-lesioned (M), 6-OHDA-lesioned (N), and

6-OHDA-lesioned L-DOPA-treated animals (O).

Distribution of GABA (PñR) is imaged in shamlesioned

(P), 6-OHDA-lesioned (Q), and 6-OHDAlesioned

L-DOPA-treated animals (R), and tyrosine

is imaged in sham-lesioned (S), 6-OHDA-lesioned

(T), and 6-OHDA-lesioned L-DOPA-treated animals

(U). There are clear differences in the

concentrations of the studied amines in different

regions of the brain and between treatments. MS

images were acquired using a MALDI-FT-ICR

mass spectrometer. Data are shown using a

rainbow scale, normalized against the total ion

count. Scale bar, 2 mm; spatial resolution =

150 mm. See also Figure S2.

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700 Neuron 84, 697ñ707, November 19, 2014 ™2014 Elsevier Inc.

and 84 pmol/mg, respectively (Figure S2); these values are

consistent with previous results obtained by HPLC (Zhao and

Suo, 2008).

Pyrylium derivatization and MS imaging were used to evaluate

the neurochemical changes that occur in 6-OHDA PD models

with and without L-3,4-dihydroxyphenylalanine (L-DOPA) treatment.

Some of the model animals were treated with deuterated

L-DOPA (D3-L-DOPA), which is converted into deuterated

dopamine (D3-DA) in vivo. The derivatization/MALDI-MS imaging

protocol successfully distinguished endogenous DA from

both the administered D3-L-DOPA and D3-DA. Sham-lesioned

rat brain tissue sections had a normal DA distribution in their

striatal regions, but the intensity of the DA signal was significantly

lower in the lesioned side of the brain, with or without

D3-L-DOPA treatment (Figures 2Añ2C). The distributions of

DA in the lesioned side of the brain and structures with low

DA levels, such as the cortex, were determined by rescaling

the intensities of the MS-derived images (Figures 2Dñ2F).

No interfering signal for D3-DA was detected in sham- or

6-OHDA-lesioned brain tissue sections (Figures 2G and 2H),

and the signal corresponding to D3-DA (derived from D3-LDOPA)

was only observed in the nonlesioned sides of brains

treated with D3-L-DOPA (Figure 2I). This is presumably due to

the destruction of dopaminergic neurons, which are essential

for the uptake and conversion of L-DOPA to DA. The distribution

images for 3-MT showed that its concentration was high

in the intact sides of the brain but lower in the lesioned sides

(Figures 2Jñ2L). The average concentration of 3-MT in the nonlesioned

sides of 6-OHDA-treated brains (Figures 2K and 2L)

was higher than that in sham-lesioned brains (Figure 2J). The

metabolic conversion of D3-L-DOPA to D3-3-MT was traced

by measuring the concentration of D3-3-MT, which was highest

in the nonlesioned side of the brain (Figure 2O); no interfering

signal was detected in untreated brains (Figures 2M and 2N).

This experiment also showed that D3-L-DOPA treatment

increased striatal GABA levels in the lesioned side of the brain

(Figure 2R) relative to those in sham- and 6-OHDA-lesioned

brain sections (Figures 2P and 2Q, respectively). Changes in

GABAergic signaling throughout the basal ganglia (including

the striatum and the substantia nigra pars reticulate) after the

destruction of dopaminergic neurons and L-DOPA treatment

have been extensively documented (Galeffi et al., 2003; Wang

et al., 2007). Tyrosine is the biosynthetic precursor of the

catecholamines and exhibited the same distribution as 3-MT

and DA (Figures 2Sñ2U).

Accurate mass FT-ICR-MS imaging was also performed on

selected regions of coronal brain sections, including the caudate

and putamen, of control and MPTP-lesioned primates (Figures

3A and 3B). The measured m/z values for selected neurotransmitters

matched the corresponding theoretical values with

sub-ppm deviations (Table S2). DA (Figure 3C) and 3-MT (Figure

3E) were localized in the caudate and putamen regions of

the brain. However, the concentrations of DA (Figure 3D) and

3-MT (Figure 3F) in the MPTP-lesioned brain sections were

around 30 and 100 times lower, respectively, than that in the

controls (Figure S3).

Several reports have discussed the varied roles of 5-HT in PD

(Fox et al., 2009). By reanalyzing the data from the experiments

used to study the distribution of DA and the effects of L-DOPA in

PD models, we were able to map the distribution of 5-HT in control

(Figure 3G) and MPTP-lesioned (Figure 3H) brain tissue sections.

5-HT was localized to the nucleus basalis (Meynert), the

hypothalamic regions, and the substantia nigra. MPTP lesioning

increased its concentration in these regions by >40% (Figure S3).

Increases in 5-HT concentrations following MPTP-induced DA

denervation have previously been demonstrated by in vivo

microdialysis; it was suggested that they are due to the brainís

compensatory processes and recovery mechanisms (Boulet

et al., 2008).

Increased levels of striatal GABA have been reported in

humans with PD, 6-OHDA-treated rats, and MPTP-treated primates.

Striatal GABA levels increased by 70% in MPTP-lesioned

brains (Figures 3I, 3J, and S3) (Boulet et al., 2008); interestingly,

its concentration also rose in the hypothalamic regions. In addition

to DA, 5-HT, and GABA, high mass accuracy FT-ICR imaging

maps were also generated for phenethylamine, tyramine, and

tryptamine (Table S2) in control and MPTP-treated primate

brains (Figure S3).

It is important to recall that while the derivatization/MALDI-MS

imaging technique makes it possible to simultaneously determine

the distributions of multiple known species by interrogating

the data set from a single experiment in a targeted manner, it

can also provide information on as-yet unidentified molecular

species.

ACh Imaging and Analysis of the Distribution of

Cholinergic Neurons Using a Deuterated MALDI Matrix

While pyrylium derivatization provides a powerful tool for

studying primary amine neurotransmitters, some important

neurotransmitters, such as ACh and alpha-GPC do not contain

primary amines and so do not react with pyrylium cations. We

therefore developed an alternative method for their imaging.

MALDI-MS analysis generally requires the homogeneous application

of a matrix to the sample surface to facilitate the laser

desorption ionization process. One of the most widely used

matrices for MALDI-MS imaging of small molecules is CHCA.

Unfortunately, all matrices generate numerous readily detected

fragments and cluster ions in the low molecular weight region

of the mass spectrum, which can mask signals from target

analytes such as ACh (m/z 146.1). One way of avoiding such

target mass interference is to use a deuterated CHCA (D4-

CHCA) matrix (Shariatgorji et al., 2012), which shifts the interfering

signals by a minimum of 4 m/z units. This method was

used to facilitate the imaging of molecular ACh and alpha-GPC

(m/z 258.1) and thereby monitor changes in their abundance

in different parts of the brain during neurological events that

affect the cholinergic system. ACh and alpha-GPC were identified

by MS/MS fragmentation (Figure S4).

Cholinergic neurons are anatomically ubiquitous. However, it

has been difficult to study their distribution in the CNS based

on the presence of ACh because of difficulties in detecting and

quantifying ACh in situ. Instead, cholinergic neuron distributions

have been studied by examining markers such as acetylcholinesterase

(the enzyme that degrades ACh), muscarinic and nicotinic

ACh receptors, and choline acetyltransferase (the enzyme

that catalyzes ACh synthesis). By using a deuterated CHCA

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matrix, we were able to directly investigate the cholinergic

neuron distribution in the brain based on the concentrations of

ACh in its various regions (Figure 4A). Imaging of rodent sagittal

sections taken from different levels of the brain revealed high

ACh concentrations in the frontal cortex, olfactory bulb, striatum,

hippocampus, medial habenula, tectum, thalamus, ventral

tegmental area, interpeduncular nucleus, pedunculopontine

tegmental nucleus, and the laterodorsal tegmental nucleus.

This is consistent with previous findings (Hammond et al., 1994).

High-Resolution MALDI-MS Imaging of ACh

High spatial resolution MALDI-MS imaging makes it possible to

map the distributions of neurotransmitters within substructures

of the brain with high spatial resolution. Using our methodology,

we determined the relative abundance of ACh in various substructures

of the hippocampus, cerebellum, and striatum at a

spatial resolution of 15 mm (Figures 4Cñ4E). The concentration

of ACh was greater in the gray matter of cerebellum than in the

white matter (Figure 4E). Certain subregions of the hippocampus

Figure 3. MALDI-MS Images of Neurotransmitters

in Coronal Primate PD Model Brain

Tissue Sections

(AñJ) MALDI-MS images showing changes in

neurotransmitter levels in one hemisphere of a

coronal brain tissue sections from a control animal

(A) and an MPTP-lesioned animal (B). MALDI-FTICR

imaging of DA (C) and 3-MT (E) in control

brains showed that these compounds are most

abundant in the caudate and putamen structures.

Conversely, both DA (D) and 3-MT (F) are almost

absent in these regions after the destruction of

their dopaminergic neurons by MPTP-lesioning.

5-HT is mostly localized in the nucleus basalis

(Meynert) of the brains, and its concentration is

significantly lower in control brains (G) than in

MPTP-lesioned tissue sections (H). The 5-HT-rich

hypothalamic structure was also imaged in control

and MPTP-lesioned brain sections, revealing

that lesioning increased its 5-HT concentration.

The concentrations of GABA in the putamen and

caudate were significantly higher in the MPTPlesioned

brain (J) than the control (I). All MS images

were acquired using a MALDI-FT-ICR mass

spectrometer. Data are shown using a rainbow

scale over a single range, normalized against the

total ion count. Scale bar, 10 mm; spatial resolution

= 200 mm. Caudate, cd; putamen, Put. See

also Figure S3.

were also enriched in ACh, notably the

granule cell layer of the dentate gyrus

(DG-sg) and the pyramidal layer (sp) of

CA1 (Ammonís horn) (Figures 4C and

4F). These subregions of the hippocampus

are delineated in Figure 4B, which

shows a MALDI-MS image generated

using data for a lipid with an m/z value

of 721.2.

The power of high spatial resolution MS

imaging was further demonstrated by

studying the distribution of ACh in the basal ganglia of the brain.

In the striatum, ACh serves as the transmitter of the large aspiny

interneurons, which have diameters of 20ñ60 mm (Kawaguchi,

1992) and comprise 1%ñ2% of the total striatal cell population

(Aosaki et al., 1998). While the cell bodies of the cholinergic interneurons

are relatively few in number, they densely innervate the

striatum with their extensive axonal arborizations (Holt et al.,

2005). As shown in Figure 4D, the striatal ACh concentrations

can be measured by MALDI-MS imaging with a spatial resolution

of 15 mm.

It is anticipated that the continuing development of MALDIMS

imaging technologies will improve to a point that enables

the analysis of small sample areas of tissue sections at subcellular

resolution. Recently, it was demonstrated that MALDIMS

instrumentation capable of imaging at resolutions < 1 mm

is technically feasible (Zavalin et al., 2012). Therefore, imaging

neurotransmitters at subcellular spatial resolution in the near

future would lead to further biological insights and make the

method even more compelling.

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Quantitative Molecular Imaging of Neurotransmitters

702 Neuron 84, 697ñ707, November 19, 2014 ™2014 Elsevier Inc.

Imaging of ACh and Alpha-GPC in Brain Sections of

Tacrine-Administered Rodents

As a final illustration of the power of the deuterated matrix technique,

it was used to investigate the distributions of ACh (Figures

5A and 5B) and alpha-GPC (Figures 5C and 5D) in sagittal sections

of mouse brains from animals that had been treated with

10 mg/kg intraperitoneal tacrine (a centrally acting cholinesterase

inhibitor) and untreated control animals. The concentration

of ACh in the tacrine-treated brains was approximately

seven times greater than that in the control brains, and the concentrations

of alpha-GPC within brain structures decreased as

their ACh levels increased. In both the control (Figure 5A) and

tacrine-treated (Figure 5B) brain sections, the ACh concentration

was determined for selected brain structures using a standard

calibration curve created by spotting known quantities of deuterated

ACh (D9-ACh) onto consecutive control brain tissue sections

(Figure S5). Absolute quantities of ACh (Figure 5E) and relative

concentrations of alpha-GPC (Figure 5F) were determined

for the whole brain, hippocampus, caudate putamen, thalamus,

and cerebral cortex regions in both control and tacrine-treated

brains. The average measured concentration of ACh was consistent

with previous results (Stavinoha and Weintraub, 1974).

Conclusions

The methods presented herein represent powerful tools for the

quantitative imaging of neurotransmitters, including biogenic

amines, amino acids, and ACh in histological brain tissue sections.

Pyrylium derivatization and deuterated CHCA matrices

extend the reach of MALDI-MS imaging, a technique with excellent

mass accuracy and resolving power, allowing it to be used in

the characterization of compounds that were previously problematic.

Pyrylium derivatization converts low molecular weight

endogenous primary amines into readily ionized species of relatively

high molecular weight. This makes it possible to simultaneously

image tyrosine, tryptamine, tyramine, phenethylamine,

DA, 3-MT, 5-HT, GABA, and Glu and other endogenous primary

amines in a single MALDI-MS experiment. The use of a deuterated

matrix allows one to sidestep problems caused by the

masking effect of matrix-derived ions in MALDI-MS analyses of

low molecular weight species, such as ACh, that lack a primary

amine group. Because MALDI-MS imaging can be performed at

a spatial resolution as low as 15 mm, these technologies make

it possible to measure the distributions of a wide range of neurotransmitters

with high spatial resolution. The concentrations

of neurotransmitters in different subregions of the brain were

quantified by comparing the intensities of their signals to those

generated by known quantities of deuterated calibration standards.

The techniques presented herein permit the simultaneous

quantitation and specific molecular imaging of multiple neurotransmitters

in situ. As such, they should find diverse and immediate

applications in fields such as neuroscience, pharmacology,

drug discovery, neurochemistry, and pathology.

EXPERIMENTAL PROCEDURES

All chemicals and reagents were purchased from Sigma-Aldrich unless otherwise

stated and were used without further purification. Ketamine was obtained

Figure 4. High-Resolution MALDI-MS Images

of ACh in a Sagittal Rat Brain Section

(AñF) MALDI-MS images of ACh in a rodent brain

sagittal tissue section. The relative distribution

and abundance of ACh determined by MS imaging

are consistent with existing data on the major

cholinergic pathways in the brain (A). ACh-rich

structures such as the HIPP, CP, CTX, and even

small structures such as the nuclei of the facial

nerves and pons are readily apparent in the

MALDI-MS image. High-resolution MALDI-MS

images (15 mm spatial resolution) of various

structures in consecutive brain tissue sections

were also acquired. Substructures of the hippocampus

were delineated by high-resolution imaging

of the distribution of a lipid with m/z 721.2

(B). The relative abundance and distribution of

ACh in the HIPP (C), a subregion of the CP indicated

by a dashed rectangular border in (A) (D),

and CB (E) were determined by high-resolution

imaging. The concentration of ACh in the gray

matter of CB is around five times greater than that

in the white matter. In the HIPP, high ACh concentrations

are apparent in DG-sg and sp regions

(F). MS images were acquired using a MALDITOF/TOF

mass spectrometer. Data are shown

using a rainbow scale and are normalized against

the total ion counts; scale bars, 2 mm, 1 mm, and

500 mm; spatial resolutions = 100 mm and 15 mm.

Caudate-putamen, CP; hippocampus, HIPP; cerebellum, CB; granule cell layer of the dentate gyrus, DG-sg; pyramidal layer of CA1, sp; cerebral cortex, CTX;

piriform area, PIR; striatum (ventral region), STRv; thalamus, TH; subiculum, SUB; stratum lacunosum-moleculare, Slm; stratum radiatum, Sr; stratum oriens, So;

cornu ammonis (field 2), CA2; cornu ammonis (field 3), CA3; polymorph layer of dentate gyrus, PO. See also Figure S4.

Neuron

Quantitative Molecular Imaging of Neurotransmitters

Neuron 84, 697ñ707, November 19, 2014 ™2014 Elsevier Inc. 703

from Parke-Davis, and xylazine was obtained from Bayer. Deuterated analogs

of neurotransmitters and phenol-2,3,4,5,6-d5 (D5-phenol) were purchased

from CDN Isotopes. Water, methanol, and trifluoroacetic acid (TFA) were obtained

from Merck.

Animal Experiments

Male Sprague-Dawley rats (approximately 150 g; Scanbur) and C57BL/6J

male mice (3 months old, Charles River Laboratories) were housed in separate

air-conditioned rooms (with a 12 hr dark/light cycle) at 20_C and a humidity of

53%. Experiments were performed in agreement with the European Communities

Council Directive of November 24, 1986 (86/609/EEC) on the ethical use

of animals and were approved by the local ethical committee at the Karolinska

Institute (N350/08).

Unilateral 6-OHDA lesioning of nigral dopaminergic axons was performed as

described previously (Zhang et al., 2008). Briefly, rats were anesthetized with

ketamine (80 mg/kg, i.p.) and xylazine (10 mg/kg, i.p.) and pretreated with desipramine

(25 mg/kg, i.p.) and pargyline (5 mg/kg, i.p.). They were placed in a

stereotaxic instrument and injected with 6-OHDA (2.5 ml of a 5 mg/ml solution)

in the medial forebrain bundle (MFB) of the right hemisphere (AP = 2.8 mm,

ML = 2.0 mm, and V = 9.0 mm).

To determine the success of nigrostriatal denervation following unilateral

6-OHDA lesioning, a single injection of apomorphine (1 mg/kg, i.p.) was administered

to the rodents 2 weeks after their treatment with 6-OHDA, and their

resulting contralateral rotations were measured over 30 min. Only rats that

performed more than 100 turns were used in subsequent studies. They were

killed by decapitation 4 weeks after the surgery, after which their brains

were removed, frozen in dry ice-cooled isopentane, and stored at 80_C until

needed.

Figure 5. MALDI-MS Images of ACh and

Alpha-GPC Concentrations in Sagittal

Mouse Brain Sections from a Control Animal

and after Administration of 10 mg/kg of the

Cholinesterase Inhibitor Tacrine

(AñD) The absolute concentration of ACh in the

control brain section (A) is about seven times lower

than that observed following the injection of tacrine

(B). Conversely, the concentration of the ACh

precursor alpha-GPC is higher in the control

sample (C) than in the tacrine-treated brain (D). The

overall levels of alpha-GPC are reduced following

tacrine treatment. The absolute concentrations of

ACh in the control (A) and tacrine-treated (B)

mouse brains were quantified using a calibration

standard curve.

(E and F) Bar graphs showing the absolute concentration

of ACh (E) and relative concentration of

alpha-GPC (F) with (black bars) and without (white

bars) administration of tacrine in whole brains and

selected structures. MS images were acquired

using a MALDI-TOF/TOF mass spectrometer. Data

are shown using a rainbow scale over a single

range, normalized against the total ion count. Scale

bar, 2 mm; spatial resolution = 100 mm. Caudateputamen,

CP; hippocampus, HIPP; cerebral cortex,

CTX; thalamus, TH. See also Figure S5.

For D3-L-DOPA treatment experiments, 4 weeks

after the surgery, rats were treated with L-DOPA

(10 mg/kg, i.p.) and benserazide (7.5 mg/kg, i.p.)

once daily for 3 weeks. Animals were sacrificed

by decapitation 30 min after the last i.p. injection,

which was deuterated (D3)-L-DOPA (10 mg/kg)

and benserazide (7.5 mg/kg). Handling and storing

the brains was performed as described above.

Primate experiments were conducted using tissue from a previously published

brain bank (Fernagut et al., 2010; Porras et al., 2012; Santini et al.,

2010). Experiments were carried out in accordance with European Communities

Council Directive of November 24, 1986 (86/609/EEC) for care of laboratory

animals in an AAALAC-accredited facility following acceptance of study

design by the Institute of Lab Animal Science (Chinese Academy of Science,

Beijing, China) IACUC. Briefly, two female rhesus monkeys (Macaca mulatta,

Xierxing; mean age = 5 ± 1 years; mean weight = 5.3 ± 0.8 kg) were used.

One animal was kept untreated as control. The other animal received daily

MPTP (0.2 mg/kg, i.v.) according to previously published protocol (Bezard

et al., 1997, 2001a, 2001b). Following stabilization of the MPTP-induced syndrome,

animal behavior was assessed as previously published (Ahmed et al.,

2010; Fasano et al., 2010; Fernagut et al., 2010; Munò oz et al., 2008; Porras

et al., 2012). The degree of parkinsonism was assessed using a validated

macaque clinical scale (Imbert et al., 2000). Animals were killed by sodium

pentobarbital overdose (150 mg/kg, i.v.), and brains were removed quickly

after death and immediately frozen by immersion in isopentane (45_C) and

stored at 80_C.

For the ACh experiments, mice that were 3 months old were injected with

saline or tacrine (10 mg/kg i.p.) and then sacrificed after 30 min according to

the procedure described above.

Tissue Section Preparation

The frozen brain tissues were cut using a cryostat-microtome (Leica

CM3050S, Leica Microsystems). Sagittal and coronal brain sections were

cut at a thickness of 14 mm. Tissue sections were transferred by thaw mounting

onto conductive indium tin oxide (ITO) glass slides (Bruker Daltonics) and

then stored at 80_C. Sections were desiccated at room temperature for

Neuron

Quantitative Molecular Imaging of Neurotransmitters

704 Neuron 84, 697ñ707, November 19, 2014 ™2014 Elsevier Inc.

15 min prior to spotting of calibration standards, after which they were imaged

optically using a photo scanner (Epson perfection V500). The samples were

then coated with matrix for ACh experiments or derivatized with or without

matrix coating for primary amine imaging experiments. To enable the quantitation

of individual neurotransmitters, deuterated ACh, DA, or GABA dissolved

in 50% methanol was spotted onto the control tissue sections to serve as

calibration standards prior to derivatization and matrix application.

Derivatization of Calibration Standards and Endogenous

Primary Amines

DPP-TFB and TMP-TFB were dissolved in methanol to prepare 1 mg/ml stock

solutions. Derivatization was performed using solutions containing 150 ml of

the stock solution of DPP-TFB or TMP-TFB in 1.5 ml of 50% methanol,

buffered by 1 ml triethylamine (TEA). The derivatization solution was applied

to the tissue sections using an automatic sprayer (ImagePrep, Bruker Daltonics).

To facilitate the derivatization reaction and to avoid delocalization of the

endogenous compounds, reagents were added using a three-step process

involving spraying, incubation, and drying, which lasted for 2.5, 15, and 50 s,

respectively. The prepared slides were then incubated for 60 min in a chamber

saturated with the vapor arising from a 50% methanol solution. To acidify the

tissue sections, 1 ml of a solution containing acetic acid, water, and methanol

(10:45:45) was sprayed over the tissue using the same setup as above. The

treated tissue sections were then incubated once more in the methanol

vapor-saturated chamber for 30 min. Primate and D3-L-DOPA-treated rat

tissue sections were analyzed using a high concentration of DPP-TFB as reactive

matrix. DPP-TFB was dissolved in 1.2 ml of methanol to prepare 8 mg/ml

of the stock solution. This solution was diluted in 6 ml of 70% methanol containing

3.5 ml of TEA. An automated pneumatic sprayer (TM-Sprayer, HTX

Technologies) was used to spray warm reagent over the dried samples. The

nozzle temperature was set at 80_C, and the reagent was sprayed (30 passes)

over the tissue sections at a linear velocity of 110 cm/min with a flow rate of

about 80 ml/min. Samples were then incubated for 15 min (dried by nitrogen

flow every 5 min) in a chamber saturated with the vapor arising from a 50%

methanol solution. Stock solutions of DPP-TFB and TMP-TFB (2 mg/ml in

methanol) and standards of DA, GABA, and Glu (1 mg/ml in methanol) were

used for in-solution derivatization experiments. Stock solutions of neurotransmitters

(2 ml) and derivatization reagents (10 ml) were added to the solvent

containing 90 ml of 50% methanol, buffered by 0.1 ml TEA. The mixture was

incubated for 30 min at room temperature, followed by addition of 1 ml of acetic

acid. Samples were spotted (0.5 ml) to dry on stainless steel plates.

Matrix Coating and MALDI-MS Imaging Data Acquisition

CHCA (0.5 ml of 10 mg/ml in 50% ACN, 0.2% TFA) was spotted over dried derivatized neurotransmitters on stainless steel plates. D4-CHCA was prepared as reported previously (Shariatgorji et al., 2012). The MALDI-MS matrix was applied to the tissue sections as follows. A 10 ml solution of 10 mg/ml

CHCA (for analysis of tissues with low concentration of derivatization reagent)

or D4-CHCA (for analysis of ACh) in 50% ACN containing 0.2% TFA

was sprayed onto the tissues sections using a TLC sprayer. The sprayer was maintained at a distance of 30 cm from the target during matrix coating, and sections were allowed to dry between coating cycles. For high spatial resolution (HSR) MALDI-MS imaging of ACh and low spatial resolution

(LSR) matrix-assisted imaging experiments, D4-CHCA and CHCA (5 mg/ml dissolved in 50% ACN containing 0.2% TFA) were applied using an automatic sprayer (TM-Sprayer, HTX Technologies). All parameters were optimized in order to reach the best sensitivity and minimize the delocalization of target compounds. The nozzle temperature was set at 98_C (for HSR experiments) and 90_C (for LSR experiments). The matrix was sprayed (eight passes for HSR and six passes for LSR experiments) over the tissue sections at a linear velocity of 110 cm/min with a flow rate of about 70 ml/min. The quantity of matrix solution applied in this way was measured using an optical sensor present in the automatic sprayer (ImagePrep, Bruker Daltonics) to ensure that the same amount of matrix was applied to each slide. All MALDI-MS imaging experiments were performed using either MALDI-TOF/

TOF (Ultraflextreme, Bruker Daltonics) or MALDI-FT-ICR (Solarix XR 12T,

Bruker Daltonics) mass spectrometer in positive ion mode using a Smartbeam

II 2 kHz laser. The mass spectrometerís operating parameters were set according to the manufacturerís recommendations for optimal acquisition

performance. The laser spot size was selected to yield intermediate and sharp levels of focus (_40 mm and 10 mm laser spot diameters) for low and high spatial resolution analyses. The laser power was optimized at the start of each run and then held constant during the MALDI-MS imaging experiment. On-tissue and on-plate MS/MS fragmentation experiments were performed in positive ion mode using the MALDI-TOF/TOF according to the manufacturerís recommended settings. Tissue sections were analyzed in a random order to prevent any possible bias due to factors such as matrix degradation or variation in mass spectrometer sensitivity. MS imaging data were visualized using FlexImaging (Bruker Daltonics), version 3.0 for

TOF/TOF data and version 4.0 for FT-ICR data. Mass spectra analysis and chemical formula calculations were performed in Data Analysis, version 4.2

(Bruker Daltonics). An in-house-developed software package was used for data processing, normalization, and quantitation. Regions of interest were manually defined in the analysis software using both the optical image and MS imaging data (Ka® llback et al., 2012). The concentration of the target compounds in defined regions of interest were calculated by correlating their signal intensities to a calibration curve derived from deuterated analogs of the analytes spotted on control tissue sections considering the brain density as 1.0 g/cm3 (Barber et al., 1970).

Autoradiographic Studies Sections that were used to study the distribution of DAT (a dopamine transporter)

were preincubated in 50 mM Tris-HCl and 120 mM NaCl (pH 7.5) for 20 min and then incubated for 1 hr in the same buffer supplemented with 50 pM [125I] RTI-55 in the presence of 1 mM fluoxetine (a selective serotonin reuptake inhibitor). The slides were washed twice for 10 s each in ice-cold

binding buffer, rapidly dipped in deionized water, and dried. The sections were then exposed to Kodak BioMax MR films for 2 days.

SUPPLEMENTAL INFORMATION

Supplemental Information includes five figures and two tables and can be found with this article online at http://dx.doi.org/10.1016/j.neuron.2014.10.011.

AUTHOR CONTRIBUTIONS

M.S. conceived the project and its essential ideas, assisted with the MALDI-MS experiments and data evaluation, and contributed to discussions.

A.N. assisted with the MALDI-MS experiments and data evaluation and contributed to discussions. N.S. conducted animal (mouse) and DAT experiments and contributed to discussions. R.J.A.G. contributed to data evaluation and discussions. X.Z. conducted animal (rat) experiments and contributed to discussions. P.K. developed the quantitation software and contributed to discussions. A.R.C and E.B. supervised the primate experiments and contributed to discussions. P.S. supervised the rodent experiments and contributed to discussions. P.E.A. supervised the overall research, conceived the project and essential ideas, assisted with the data evaluations, and contributed to discussions. All authors contributed to the writing of the manuscript.

ACKNOWLEDGMENTS

The authors acknowledge Dr. Matthias Witt and Patrik Ek (Bruker Daltonics) for assisting with FT-ICR-MS imaging and Dr. Andras Szabo and members of Ubichem laboratory for synthesis of deuterated matrix. This work was supported by the Swedish Research Council (Medicine and Health 2013-

3105, Natural and Engineering Science 2010-5421, and Research Infrastructure 2009-6050 to P.E.A.), AstraZeneca R&D, UK (to P.E.A. and A.N.), Agence Nationale de la Recherche (ANR-12-BSV4-0001-01 to E.B.), and the Michael J. Fox Foundation (to E.B.).

Accepted: September 29, 2014

Published: November 6, 2014

Neuron Quantitative Molecular Imaging of Neurotransmitters Neuron 84, 697ñ707, November 19, 2014 ™2014 Elsevier Inc. 705

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Social work practice in mental health

Social work practice in mental health
Social work practice in mental health

Social work practice in mental health

The domain of social work in mental health is that social context and social consequences….discuss.

Your assignment must follow these formatting requirements:

  • Be typed, double spaced, using Times New Roman font (size 12), with one-inch margins on all sides; citations and references must follow APA or school-specific format. Check with your professor for any additional instructions.
  • Include a cover page containing the title of the assignment, the student’s name, the professor’s name, the course title, and the date. The cover page and the reference page are not included in the required assignment page length.

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Abnormal Psychology-Substance Use Disorder

Abnormal Psychology-Substance Use Disorder
Abnormal Psychology-Substance Use Disorder

Abnormal Psychology-Substance Use Disorder

The Case Study is: SUBSTANCE USE DISORDER (Case study 4)

(1) Explain what the purpose of a clinical assessment is.

(2) Identify a commonly used instruments clinicians or psychologists use to screen for a particular disorder and why it would be used. Note, the DSM-5 is not a screening instrument. An instrument helps to identify if a patient is experiencing the symptoms listed in the DSM-5. An example of an instrument is the Beck Depression Inventory that screens for symptoms Major Depressive Disorder, or the PC-PTSD that is designed to screen for post- traumatic stress disorder.

(3) During a clinical assessment, why would a clinician consider the level of severity of a patient’s symptoms?.

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Mental health disorders Essay Assignment

Mental health disorders
Mental health disorders

Mental health disorders

how does the media represent/view offenders (pick one type of crime) with mental health disorders or those who consume illegal substances”

SUBJECT: Criminology, ”how does the media represent/view offenders (pick one type of crime) with mental health disorders or those who consume illegal substances” (this is a working title and i do not have a problem with it being tweaked as long as it is of a criminology nature)

my course requires me to complete a PowerPoint poster of 1000 words which is needs the findings of the research put in another format i have included all the things which i think may be useful.

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Abnormal Behavior Research Paper Assignment

Abnormal Behavior
Abnormal Behavior

Abnormal Behavior

This Abnormal Behavior paper is needed for my Mental Health in Criminal Justice class.
Write a 700- to 1,050-word paper in which you examine the field of abnormal psychology. Address the following items:
• Describe challenges related to classifying normal versus abnormal behavior.
• Analyze the psychosocial, biological/medical, and sociocultural theoretical models related to the development of abnormal behavior.
• Describe the relationship between abnormal behavior and criminal behavior.
• Explain how correctional institutions have been affected by abnormal behavior.

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Moderate Schizophrenia Research Assignment

Moderate Schizophrenia
                Moderate Schizophrenia

Moderate Schizophrenia

For each of the following scenarios, write at least 150 words in response. Your short answer response must also include 1 We can write this or a similar paper for you! Simply fill the order form!.

1. Alyssa’s husband has passed away. As a result of this tragedy, she is suffering from acute social anxiety. She stays in her house all day by herself. The only place she went after her husband’s death was his funeral. Months later, shockingly, you see her at the grocery store. Which approach is the best way to support Alyssa and why?

2. Your best friend, Brandon, has recently lost his job. Coincidentally, two weeks ago, you also lost your job and quickly found another job. Brandon comes to you for advice because he found out about your situation through a mutual friend. What is the best way to support Brandon and why should you take this approach?

3. As a young child, Kara’s parents abused her. After being adopted at the age of 6, she was quickly found to be suffering from moderate to severe depression. Kara is now a student in your second-grade class. She hardly ever speaks and has lots of trust issues. How can you help Kara’s situation and why should you take this approach?

4. Sean has been diagnosed with moderate Schizophrenia. In an effort to improve his ability to socialize appropriately, he and a group of other schizophrenics are invited to your workplace for one week to work on this skill as part of their rehabilitation. At lunch, surprisingly, Sean asks to sit with you. What should you say to support Sean and why should you take this approach? my instructor wants me to answer these questions

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Disorders of Aging and Cognition Essay Paper

Disorders of Aging and Cognition
   Disorders of Aging and Cognition

Disorders of Aging and Cognition

5-7 page paper on abnormal psychology about disorders o aging and cognition.
Talking about older and elderly people, Alzheimer’s disease, Parkinson’s, stress,
depression, anxiety disorders, their mental health, clinicians discovering the elderly, sports head injuries, chronic traumatic encephalopathy, psychotic disorder, substance misuse, etc. Please stick to these topics and follow the directions written by my teacher attached in the file.

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  • Be typed, double spaced, using Times New Roman font (size 12), with one-inch margins on all sides; citations and references must follow APA or school-specific format. Check with your professor for any additional instructions.
  • Include a cover page containing the title of the assignment, the student’s name, the professor’s name, the course title, and the date. The cover page and the reference page are not included in the required assignment page length.

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