1

Neuropeptides
CHAIRPERSON-DR.SHANKAR.K
PRESENTER –DR.SWATHI
Overview 2

 Introduction

 Biosynthesis of neuropeptides

 Distribution and Regulation

 Neuropeptide signaling

 Degradation and functions of neuropeptides

 Specific neuropeptides

 Conclusion

 References
Introduction 3

 David de Wied (1925–2004) coined the term neuropeptide.

 David de Wied formulated the hypothesis that peptides localized

within neurons may directly influence brain function and consequently
human and animal behaviour.

 A neuropeptide is a small proteinaceous substance produced and

released by neurons through the regulated secretory route and acting
on neural substrates.
Introduction 4

Criteria for Neuropeptides

 The hallmarks of neuropeptides are

(1) gene expression and biosynthesis by neurons,

(2) storage and regulated release upon demand, and

(3) the ability to modulate or mediate neural functioning directly

through neural receptors.
Introduction 5

 A neuropeptide is a chain of two or more amino acids linked by

peptide bonds and differs from other proteins only in the length of
the amino acid chain.

 Range : 2 - >40 amino acids

 Peptides with >90 amino acids – proteins.

 Over 100 unique biologically active neuropeptides have been

identified in the brain
Introduction 6

Classification - Neuropeptides can be classified into following categories

1) Opioid neuropeptides (met-enkepalin, leu- enkephalin,-endorphin, dynorphin),

2) Gut-brain peptides (substance P, cholecystokinin, galanin, bombesin,

neurotensin, neuropeptide Y),

3) Hypothalamic releasing hormones (corticotropin-releasing factor, hypocretin,

melanin-concentrating hormone (MCH)),

4) Pituitary hormones (AVP, ACTH),

5) Miscellaneous peptides (bradykinin).

Biosynthesis 7

 Biosynthesis of neuropeptides occurs according to the classical

steps described for protein synthesis:
Transcription of an mRNA from a specific gene

Translation of a polypeptide preprohormone encoded by that mRNA

Posttranslational processing

Pre-prohormone
Biosynthesis 8
mRNA encoding the preprohormone
R
E
R
Polypeptide is translated into the cisternae of the RER
R
E
R
Signal peptide anchored in the RER membrane
signal endopeptidase

PACKAGED INTO VESICLES

The prohormone polypeptide GOLGI
9
Biosynthesis 10

 After storage in the large vesicles, neuropeptides are released

through exocytosis. This occurs when an action potential arrives
down the axon in the nerve terminal, causing a calcium influx
through calcium channels.

 The increased calcium concentration causes a cascade of

molecular events leading to exocytosis.
Distribution and Regulation 11

 Peptides involved in regulating pituitary secretion are concentrated

in the hypothalamus

 Other brain regions - limbic structures, cerebral cortex, midbrain,

hindbrain and spinal cord

 Peptides are colocalized and released with other neuropeptide or

nonpeptide neurotransmitters

 Peptides are cotransmitted with nonpeptide neurotransmitters and

modulate neurotransmission of both the pre- and postsynaptic cells.
Neuropeptide signaling 12

 Neuropeptide neurotransmitters are typically released from axonal

terminals into a synapse, where binding to neuropeptide receptors
changes the postsynaptic membrane potential, either depolarizing or
hyperpolarizing the cell.

 The cellular signaling of neuropeptides is mediated by specific

neuropeptide receptors.
Neuropeptide signaling 13

 Neuropeptide neuromodulators and neurohormones act through

modulation of second messenger pathways.

 Majority of neuropeptide receptors - GPCR belonging to the same

family of proteins as the monoamine receptors.
Neuropeptide signaling 14

 Most common types of receptor signaling pathways involve the

activated G protein modulating the activity of either adenylate cyclase
(e.g., Gs or Gi) or PLC (e.g., Gq).

 Stimulation of adenylate cyclase results in increased cyclic adenosine

monophosphate (cAMP) production in the cell, while stimulation of
PLC results in increased diacylglycerol (DAG) and inositol
triphosphate (IP3).
Neuropeptide signaling 15

 Many neuropeptides appear to be coreleased with at least one other

neurotransmitter at the synapse

Example : NPY is coreleased with norepinephrine from same vesicles

within cells to act simultaneously on the target cell

Degradation of neuropeptides 16

 The neuropeptide is catabolized to metabolites in the synaptic cleft by

catabolic peptidases.

 There are two broad categories of peptidases: exopeptidase and

endopeptidase.

 The enzymes may be found bound to post- or presynaptic neural

membranes or in solution in the cytoplasm and extracellular fluid, and
they are distributed widely in peripheral organs, and serum, as well as
in the CNS
Degradation of neuropeptides 17
Functions of neuropeptides 18

 Thermoregulation

 Food and water consumption

 Sex, sleep, learning, memory, locomotion

 Responses to stress and pain

 Emotion and social cognition

Differences With Classical Neurotransmitters 19

 Like the “classical” small molecule neurotransmitters,neuropeptides function

as chemical mediators enabling neuronal communication.

 However, contrary to such classical neurotransmitters, neuropeptides differ in

their features.
20
Investigating neuropeptide function 21

Levels of analysis :

1. Molecular structure & biosynthesis of peptide and receptor

2. Regulation of expression and release of peptide

3. Behavioural effects of peptide

Investigating neuropeptide function 22

Identification of structure

1. Chemical analysis of purified biologically active peptides – cloning

and characterization of genes

2. Gene structure – molecular regulation and chromosomal localization

– ultimately aids in examining the peptide’s potential in pathogenesis

3. Structural characterization – production of immunological and

molecular probes to determine distribution and localization of
peptides
Investigating neuropeptide function 23
Investigating neuropeptide function 24

Other techniques

1. siRNA, shRNA techniques - selective degradation of targeted mRNA in

specific brain regions – study function of specific neuropeptide producing
neurons

2. CRISPR/Cas system - gene editing - separate temporal effects of

up/downregulating neuropeptides and genes

3. Optogenetics : LASER light to activate neuronal firing- provide special and

temporal resolution of real-time neuronal activation in different models, in
combination with the modulators and technologies
Neuropeptides in psychiatric diseases 25

 The involvement of neuropeptides in psychiatric diseases may

result from different non exclusive mechanisms, occuring at both
cellular and systemic levels.

 Cellular level - Either the neuropeptide neuron is directly involved

in the disease, or it is involved because of its interaction with
neurons expressing a classical neurotransmitter known to be
involved in the disease.
26
Neuropeptides in psychiatric diseases 27

 Systemic level-

1) The neuropeptide is directly responsible for the disease, either

because it participates in its etiology,or because its dysregulation
induces a core symptom of the psychiatric disease. Abnormalities in
the neuropeptide may produce a core symptom, leading to a given

psychiatric illness including several Symptoms.

Neuropeptides in psychiatric diseases 28

2) Neuropeptides may not only modulate the core symptom per se

or the etiology of the disease but some specific related symptoms,
such as sleep, motivation or pain .
29
Specific Neuropeptides 30

1. Thyrotropin-releasing hormone (TRH)

2. Corticotrophin releasing factor (CRF)

3. Oxytocin (OT)

4. Arginine vasopressin (AVP)

5. Neurotensin (NT)
Thyrotropin-Releasing Hormone 31

 1969 - TRH - pyroglutamyl histidyl prolinamide tripeptide - first of the

hypothalamic releasing hormones to be isolated.

 Nobel prize for Dr.Andrew Schally and Roger Guillemin in 1977 for the
same

 Discovery of structure - demonstrated peptide hormones secreted

from the hypothalamus regulate secretion of hormones from the
anterior pituitary

 Location : chromosome 3q13.3-q21

 Hypothalamic TRH neurons 32

Median eminence

Release TRH
(hypothalamo-hypophyseal
portal system)

Adenohypophysis

TSH into the systemic circulation

Thyrotropin-Releasing Hormone 33

 TRH immunoreactive neurons are located in olfactory bulbs,

entorhinal cortex, amygdala, hypothalamus, midbrain structures.

 TRH receptor is a member of the seven-transmembrane domain,

GPCR family

 Within CNS : TRH modulates dopamine, serotonin, acetylcholine

and opioids
Thyrotropin-Releasing Hormone 34

 TRH mRNA expression in the PVN of the hypothalamus is

decreased in patients with major depression

 TRH - arouses hibernating animals, counteracts the behavioral

response and hypothermia produced by a variety of CNS
depressants

 CSF TRH concentrations are elevated in depressed patients as

compared to controls supports hypothesis of TRH hypersecretion
with no clear information of regional CNS origin
Corticotropin-Releasing Factor and 35
Urocortins
 1950s - pituitary extracts contain a factor,crf, which could
stimulate the release of ACTH from anterior pituitary cells in vivo

 Vale and colleagues isolated and characterized CRF as a 41-

amino acid peptide in 1981

 Located on chromosome 8q13 and has 2 exons

 Related neuropeptides, urocortin 1, urocortin 2, and urocortin 3 -

share similar gene structures
Corticotropin-Releasing Factor and 36
Urocortins
 Distribution : widely distributed throughout the brain

 Sites : PVN of hypothalamus, raphe nuclei and locus coeruleus

(LC).

 CRF is the primary hypothalamic ACTH secretagogue .

 It acts as an extra hypothalamic neurotransmitter/neuromodulator

in a CNS network which, along with the urocortins, globally
coordinates responses to stressors
Corticotropin-Releasing Factor and 37
Urocortins
 The physiological and behavioral roles of the urocortins are less
understood.

 Urocortin 2 and 3 mediate anxiolytic action and may dampen the

stress response.

 This has led to the hypothesis that CRF and the urocortins act in
opposition, but this is likely an oversimplification
Corticotropin-Releasing Factor and 38
Urocortins
 Urocortin 1 is primarily synthesized in the Edinger–Westphal nucleus,
lateral olivary nucleus, and supraoptic hypothalamic nucleus.

 Urocortin 2 is synthesized primarily in the hypothalamus.

 Urocortin 3 cell bodies are found more broadly in the extended

amygdala, perifornical area, and preoptic area.
Corticotropin-Releasing Factor and 39
Urocortins
 The effects of CRF and the urocortins are mediated by at least two
receptor subtypes, CRF1- and CRF2-receptor.

 The urocortins have been proposed to be the endogenous ligands for

the CRF2 receptor
Corticotropin-Releasing Factor and 40
Urocortins
Corticotropin-Releasing Factor and 41

Urocortins
 Hyperactivity of the HPA axis in major depression remains one of the
most consistent findings in biological psychiatry.

 The reported HPA axis alterations in major depression include

 hypercortisolemia,

 resistance to dexamethasone suppression of cortisol secretion (a

measure of negative feedback),
Corticotropin-Releasing Factor and 42

Urocortins
 blunted ACTH responses to intravenous CRF challenge,

 increased cortisol responses in the combined dexamethasone/CRF

test, and elevated CSF CRF concentrations

 Normal subjects : CRF stimulation test (i.v or s.c) yields robust ACTH,
β-endorphin, β-lipotropin, and cortisol responses.
Corticotropin-Releasing Factor and 43
Urocortins
 In patients with major depression, blunting of ACTH or β-endorphin
secretion with a normal cortisol response has been repeatedly
reported

 Early-life stress apparently sensitizes the HPA axis and leads to a

greater risk of developing depression later in life.
Oxytocin and Vasopressin 44

 Vasopressor effects of posterior pituitary extracts - first described in

1895, and given the name vasopressin

 1953- OT became first peptide hormone to be chemically synthesized,

leading to the Nobel Prize in chemistry – to Vincent du Vingeaud in
1955.

 Human OT and AVP genes – Tandemly situated on chromosome 20p13

 Structural composition : cyclical nonapeptides with a cysteine–cysteine

disulfide bond and differ at two amino acid residues.
Oxytocin and Vasopressin 45

 Oxytocin and vasopressin mRNAs - the most abundant

messages in hypothalamus

 Location : magnocellular neurons of paraventricular and

supraoptic nuclei of hypothalamus

 These nuclei send axonal projections to the neurohypophysis

and produce all the OT and AVP that is released into
bloodstream
Oxytocin and Vasopressin 46

 OT released from the pituitary - functions associated with female

reproduction (uterine contractions and milk ejection reflex )

 AVP also known as antidiuretic hormone - regulates water

retention in the kidney and vasoconstriction through interactions
with vasopressin V2(not in brain) and V1a receptor subtypes
Oxytocin and Vasopressin 47

 V1a receptor is distributed widely in the CNS - mediate most of its

behavioral effects

 V1b receptor is concentrated in anterior pituitary and hippocampus - V1b

knockout mice display decreased aggression and altered social behaviors

 AVP acts to potentiate the effects of CRF on ACTH secretion through V1b
receptors (colocalised with CRF in parvocellular neurons)

 Microdialysis experiments: AVP is released within the CNS in response to

stressful stimuli
Oxytocin and Vasopressin 48

 OT has been reported to facilitate

1. Female sexual behavior

2. Increase social interest

3. Onset of maternal behavior

 Studies in knockout mice - OT may be responsible for

processing socially salient stimuli
Oxytocin and Vasopressin 49

 Studies in highly social, monogamous rodents - OT is

involved in forming of selective social attachments
between mates

 Extra hypothalamic AVP is predicted to be involved in regulation of sex-

specific behaviors in males

 AVP facilitates affiliation and social attachment in monogamous mammals

 Three genetic association studies reported associations between

polymorphisms in V1aR promoter and ASD.
Neurotensin 50

 Isolated in 1973 based on its hypotensive properties from bovine

hypothalamus

 Neurotensin-neuromedin N gene - originally cloned from canine

ileal mucosa

 Human NT gene - chromosome 12 (12q21)

 NT-producing cells- midbrain (VTA ,substantia nigra), ventral

striatum, extended amygdala, lateral septum, and arcuate
nucleus
Neurotensin 51

 Location : midbrain- VTA and SN.

 Within the VTA - found in dense-core vesicles only in tyrosine-

hydroxylase–positive staining cell bodies, indicating
colocalization with dopamine.

 NT-dopamine cells project to the prefrontal cortex, striatum,

amygdala, and lateral septum.
Neurotensin 52

Neurotensin receptors

 Three receptors – NT1, NT2, NT3.

 NT1 and NT2 receptors are seven-transmembrane, GPCRs.

 NT3 is a Type I amino acid receptor with a single

transmembrane domain and intracellular in location.
Neurotensin 53

Neurotensin has gained interest in research on the pathophysiology of

Schizophrenia because

1. NT system – positioned anatomically to modulate neural circuits

involved in schizophrenia

2. Peripheral administration of antipsychotic drugs has been shown to

consistently modulate NT systems

3. Evidence of central NT systems being altered in schizophrenic

patients
Neurotensin 54

 Both antipsychotic drugs and NT neurotransmission enhance

sensorimotor gating, as sensorimotor gating defect is seen in
Schizophrenia.
Cholecystokinin 55

 CCK was originally discovered in the gastrointestinal tract.

 Location- cortex, striatum, hypothalamus, hippocampus, and

amygdala.

 Colocalized with dopamine in the VTA

 Functions -decreases dopamine release

Infusions of CCK induce panic in healthy individuals

Pentagastrin (agonist) - HR,BP and physical symptoms

of panic
Substance P(receptor- NK1) 56

 Location - Amygdala, hypothalamus, periaqueductal gray, LC, and

parabrachial nucleus.

 Colocalized with norepinephrine and serotonin

 Implications- Pain neurotransmitter

 Both depressed and PTSD patients exhibit elevated CSF

substance P concentrations.

 NK1 antagonist is more effective than placebo, equal to paroxetine

in patients with major depression
Neuropeptide Y ( NPY ) 57

 NPY is a 36 amino acid peptide

 Location- Hypothalamus, brain stem,Spinal cord, and several

limbic structures

 NPY is colocalized with serotonergic and noradrenergic neurons.

 Functions- involved in the regulation of appetite, reward, anxiety,

and energy balance.
Neuropeptide Y ( NPY ) 58

Implications-

 Containment of negative effects following exposure to stress

 Decreased in depression

 Plasma NPY elevated in soldiers subjected to the

“uncontrollable stress” of interrogation
Conclusion 59

 Neuropeptides are neurotransmitters made up of amino acids, each

connected by peptide bonds.

 They are relatively large and are composed of 3 to 36 amino acids.

 They are released into the synaptic cleft along with another
neurotransmitter and are degraded by peptidases.

 The study of these neuropeptides and their receptors has increased

considerably our understanding of their mechanisms of action in a
wide range of physiological and pathophysiological situations.
References 60

 Shaun P. Brothers, Larry James Young, and Charles B. Nemeroff,

Neuropeptides: Biology, Regulation, and Role in Neuropsychiatric Disorders
Kaplan and Sadock’s Comprehensive textbook of Psychiatry,10 th edition,
Wolters Kluwer.

 Brain SD, Cox HM. Neuropeptides and their receptors: innovative science
providing novel therapeutic targets. British journal of pharmacology. 2006 Jan
1;147(S1):S202-11.

 Burbach JP. What are neuropeptides?. InNeuropeptides 2011 (pp. 1-36).

Humana Press.
References 61

 Belzung C, Yalcin I, Griebel G, Surget A, Leman S. Neuropeptides in

psychiatric diseases: an overview with a particular focus on depression
and anxiety disorders. CNS & Neurological Disorders-Drug Targets
(Formerly Current Drug Targets-CNS & Neurological Disorders). 2006 Apr
1;5(2):135-45.

 Nillni EA. The cell biology neuropeptide hormones. InTextbook of Energy

Balance, Neuropeptide Hormones, and Neuroendocrine Function 2018
(pp. 109-139). Springer, Cham.
62