The characterization of the neurobiology of psychiatric diseases

Studies in humans:

Problems associated with diagnostic, heterogeneity of population and effects of

medication.

Difficulties to know whether alterations precede (vulnerability factor) or are the

consequence of the pathology (or medication).

Behavioral and neuropsychological tests.

Genetic markers.

Peripheral Biomarkers.

Electroencephalografy (including evoked potentials and magnetoencephalography)

Neuroimage (structural and functional).

Post-mortem biochemical or histological analyses

Development of animal models

 Neuroimaging
 Different techniques: MRI, fMRI, PET and SPET.

 MRI
 Estructural/volumetric changes
 Axonal connectivity (functional anisotropy)

 fMRI gives no absolute values but it is based on comparison between

different times (or situations):
 Differential activation.
 Functional connectivity

 Spatial resolution can be better with fMRI than with the others.

 Theoretical (lack of consensus) and practical (atlas) problems related

to the delimitation of the different areas.

 It is important to consider:
 A) The number and type of patients.
 B) The mood state at the time of scan
 C) Whether or not the subjects are taking medication
 Depressive disorders and DSM-V

 Classification

• Disruptive mood dysregulation disorder

• Major depressive disorder
• Persistent depressive disorder (previously chronic depressive disorder
and dysthymia): mood disorders for at least 2 years.
• Premenstrual dysphoric disorder
• Substance/medication-induced depressive disorder
• Others

 Prevalence is in general about 13-23%, 7% for major depressive disorder. 2-3

fold higher in females versus males.

 Age of onset typical, in some cases, during adolescence

 Moderate genetic contribution (2-3 fold probability increase in families)

 Criteria for major depressive disorder (DMS-V)

• A. Five or more of the following symptoms:

• Depressed mood most of the day, nearly every day
• Markedly diminished interest or pleasure in almost all activities
• Significant weight loss or gain associated with reduced or increased appetite
• Psychomotor agitation or retardation
• Fatigue or loss of energy
• Feelings of worthelessness or excesive/inappropriate guilty
• Diminished ability to think, concentrate and take decisitions.
• Recurrent thoughs of death (not just fear of dying) or suicidal ideation.

• B. The symptoms cause clinically significant distress or impairment in

daily life.

• C. The episode has not substance or organic origin.

• D. The episode is not better explained by other psychiatric pathologies.

• E. No manic o hypomanic episode

 Anatomy of prefrontal cortex

There is no consensus about delimitations of the different prefrontal areas in humans and the
precise relationship with other primates.

The relationship with the rat is still more difficult to establish

Ray & Zald, Neurosci Biobehav Rev 36, 479, 2012

sgACC, pgACC y aMCC have important
connections with the amygdala (basolateral
complex), PAG and hypothalamus, whereas
aMCC y pMCC have connections with lateral
PFC and premotor areas.

Sucortical connections of the ACC (v.g. with

the amygdala) are far greater than those of
the mPFC.

It appears that rostral parts of CC are more

emotional whereas the posterior parts are
more cognitive, but the boundaries are not
clear and the functions are likely not to be
clearly segregated.
 Neuroanatomical and functional alterations in depression

• Reduced hippocampal volume preceding the pathology that can be further reduced over
the course of the pathology.

• Reduced gray matter in the PFC and ACC

• Impaired insula functional recognition of disgust

• Hyperactivity of sgACC (area 25) and amygdala

• Altered functioning of the default system (at rest, in absence of sensorial stimulation)
– Enhanced activity of vmPFC/ACC
– Hypoactivity of DLPFC
– Impaired connectivity

• Depressives are not impaired in early (inconscious) processing of negative valence

materials) but rather in further conscious processing and disengagement from negative
stimuli.

• Exacerbated response to negative feedback

• Impaired response to reward (wanting not liking)

 NIH alternative to DSM criteria: RDoC

Morris SE and Cuthbert BN (2012) Dialogues in Clinical Neuroscience, 14, 29-37

Pryce CR and Seifritz E (2011) Psychoneuroendocrinology, 36, 308-329
 Cognitive theories of depression

• Initially proposed by Aaron T. Beck, posits that prepotent maladaptive schematic

representations of the self, world and future are activated by life experiences leading
to preferential conscious processing bias and dominance of negative and threat-
related thoughts and interpretations.

• This hypothesis has lead to corresponding cognitive therapy (CT).

• Depression is associated with cognitive deficits and some theories argue that
the relationship is bidirectional: positive mood promotes associative processing and
associative processing promotes positive mood.

• Rumination around a narrow focus might acts in a opposite way to broad

associative processes and the medial prefrontal cortex connections to the medial
temporal lobe play a critical role

• Improvement of symptoms after CT for anxiety and depression is associated with

decreased activity in sgACC and amygdala-hippocampal regions and increased
activity in mPFC, OFC and ACC
 Classical theories about biological bases for depression

• Monoamine hypothesis
• Dopamine anhedonia
• Noradrenaline antidepressants
• Serotonin antidepressants

• The influence of stress

• Altered activity of the hypothalamic-pituitary-adrenal axis

• Altered biological rhythms

• Stational depression
• Circadian alterations in mood
• Therapeutic effects of sleep deprivation
 Monoaminergic hypothesis

• Reduced 5-HT and 5-HIAA in CFS (mainly in suicides).

• Altered reuptake of serotonin in platelets.

• Reduced prolactin response to serotoninergic drugs

• Association with polymorphism in 5-HT related genes (i.e. short and long
forms of the 5-HT transporter (5-HTT).

• Action of IMAOs

• Action of SSRI

• Our knowledge about serotoninergic brain system

Antidepressants (ADs):
- MAO inhibitors
- Inhibitors of noradrenaline and serotonin reuptake (tryciclics))
- Serotonin Selective Reuptake Inhibitors (SSRI)

The therapeutic effects of ADs are related to chronic, not acute, biochemical effects

Classical ADs have important

side-effects (dirty drugs)
Delayed action of antidepressants can be explained by desensitization of
autoreceptors
(Rausch et al, Neuropsychopharmacology, 32, 2274, 2006)
 Depression might not be related to alterations in monoamines

• Blockade of catecholamine synthesis causes a fast relapse in those patients who were treated
with noradrenaline reuptake inhibitors and responded to treatment, but not in those who were
treated with serotonin reuptake inhibitors.

• The opposite is observed with the blockade or reduction of serotonin synthesis (tryptophan
depletion).

• In some cases tryptophan depletion causes signs of depression in remitted patients not treated
previously with antidepressants
 Relationship with the hypothalamic-pituitary-adrenal axis

• High levels of CRH in CSF

• Reduced prefrontal cortex CRH receptors
• Higher number of CRH+ neurons in the PVN

• Reduced ACTH response to CRH

• Hypercortisolemia
• Impaired glucocorticoid (dexamethasone, DEX) negative feedback
• Enhanced response to the DEX+CRH test

• Opposite effects of antidepressants and stress regarding:

– Behavioral consequences of stress
– Resting and acute stress levels of HPA hormones
– Enhanced negative feedback
– Enhanced traslocation of GR to the nucleus
 Depression is associated to
central and peripheral HPA
hyperactivity

 Chronic high levels of cortisol

(e.g. Cushing) can induce
depression

• HPA hyperactivity is not

observed in all patients.

• Hypercortisolemia and DEX

resistance can be dissociated
Biological rhythms and depression
Monteleone P, Maj M, Eur Neuropsychopharmacol
18, 701 (2008)

Palagini et al. Sleep Med Re 17, 377 (2013)

Sleep deprivation has rapid but short lasting effects on mood in some subjects
(partial sleep deprivation or REM sleep deprivation are less effective)
Recent hypotheses about depression: BDNF
 BDNF

 Met-Met mice showed reduced hippocampal volumen, reduced dendritic

arborization and impaired synaptic plasticity

 However, changes in BDNF might exert different effects depending on the

brain regions (v.g. hippocampus versus tegmental ventral area-VTA)

 In humans reduced plasma BDNF levels has sometimes observed in

psychiatric diseases, including depression

 Availbility of TrB agonists (7,8-dihydroxyflavone) can allow

pharmacological studies in humans
 Neurogenesis, depression and antidepressants

- Reduction of neurogenesis does not result in depression-like behaviour.

- Enhanced neurogenesis is involved in some, but not all, effects of

antidepressants in animal models.
 Depression is associated with pro-inflammatory states

Pace et al, Am J Psychiatry 163:1630, 2006

Kling et al, Biol Psychiatry 62, 309, 2007
Depression as a low, chronic, inflammatory process
NLRP3: multiprotein
complex of glial cells that
responds to PAMPs and
DAMPs
Inflammasome: the link between Depression and cardiovascular diseases?
Alcocer-Gómez & Cordero, Nat Rev Cardiol 2017
 Antidepressive therapies
• Cognitive therapies
• Pharmacological treatments
– Classical ADDs
– Rapidly acting drugs
– Other putative drugs (i.e. CRH antagonists)

• Combination of cognitive and pharmacological therapies

• Electroconvulsive shocks

• Transcraneal magnetic stimulation

• Deep Brain Stimulation (experimental, sgACC, Cg25)

Depression and synaptic changes in the prefrontal cortex

Abdallah GS et al Annu Rev Neurosci 66, 509 (2015)

 (low, subanesthetic doses)

Different mechanisms, in addition to enhanced glutamate release (the latter one in Figure),
has been proposed to explain the rapid action of ketamine
Animal models of depression
Main criteria to develop animal models:

 Construct validity: similar etiology and putative underlying processes.

 Face validity: similar behavioral changes (symptoms).

 Predictive validity: similar efficacy of treatments (e.g. drugs)

 Objectives of developing animal models of depression

¿Which type of depression are we trying to model?

¿Which are the purposes of the model:

 To search for or to evaluate antidepressant treatments

 To study biological bases of depression

Pure pharmacological models evaluate how putative antidepressant drugs can interfere
with the effects of selected drugs:

-Atenuation of hypothermia or sedation caused by reserpine

-Potentiation of L-DOPA-induced aggression

-Potentiation of amphetamine-induced anorexia

-Potentiation of the serotoninergic syndrome caused by 5-HTTP (e.g. tremor, head-twicht)

These models has low validity and only indentify new drugs with expected mechanisms
of action.
 Evaluation of depression-like behavior

 Motivation to explore novel environment

 Social interaction, including dominance-subordinate relationships

and social avoidance

 Coping strategies in inescapable situations: active versus passive

 Reward:

 Intracraneal self-stimulation

 Stimulus-induced dopamine release

 Conditioned place preference in response to rewarding stimuli

 Preference for sweet solutions (sucrose, saccharin)

 Effort-related performance

 Cognitive bias
 Forced swimming test (Porsolt et al, Nature 1978)

The test actually evaluate the coping

strategy of the animals when facing
inescapable situations.

Antidepressants (including REM sleep

deprivation) reduce immobility (or
increase escape).

Individual differences in the time spent

active or immobile might be useful to
identify animals prone to develop
depression-like behavior.

The most active

escape behavior
(struggling)
 Previous experience of animals with the forced swim situation and the
effects of diazepam and DMI

DAY 1 DAY 3 DAY 5

 Models having marked similarities with the classical forced swim test

Tail-suspension test
 Main animal models to induce depression-like behavior

 Environmental changes

 Separation from mothers and peers (primates)

 Learned helplessness

 Social defeat

 Chronic unpredictable/variable stress (CUS; CVS)

 Drug withdrawal

 Olfactory bulbectomy

 Genetic models

 Genetically selected animals

 Genetically modified animals

 Exposure to uncontrollable electric tail-shocks: the learned
helplessness paradigm
(Seligman, Overmier and Maier)
Escapable shock, ES Inescapable shock, IS
The degree of control over the tail-shocks DID NOT ALTER the response
of the HPA axis

Maier et al, Behav Neurosci 100, 669, 1986

 The inescapable foot-shock exposure model has important differences
with the tail-shock model

Development of context fear conditioning and enhanced freezing that impaired avoidance/escape
(failures and latency).

These negative consequences of stress are reduced by chronic antidepressant treatment given
before the first exposure to inescapable shocks or before the avoidance/escape task.
 Chronic Stress Models
Different chronic stress models:

 Chronic intermittent and repeated stress.

 Chronic intermittent variable (unpredictable) stress.

 Chronic intermittent social stress (daily repeated social defeat)

 Chronic continuous stress (i.e. superpopulation, social stress)

Initially, most studies have focused on the two first models, but there
is increasing interest for social defeat/social stress models
Changes caused in the HPA axis by chronic exposure to high intensity stressors

The observed changes depend on factors such as the intensity and length of exposure
to the stressors. However, under conditions of certain severity we can observe at
least some of these changes:

• Increased adrenal weight and maximum adrenocortical response to ACTH

• Increased in resting levels of corticosterone at lights on

• Increased expression of POMC in the anterior pituitary

• Reduction in the number of CRF receptors in the anterior pituitary, BUT increased
responsiveness of ACTH to exogenous administration of CRF

• Increased expression of CRF y AVP in the PVN

• Reduced number of glucocorticoid type II receptors (GR), but not usually of type I
(MR) in the hippocampal formation
Variable or unpredictable chronic stress (CUS)

This model was initially proposed by Katz

RJ et al (1981) and popularized by Willner.

Katz et al. Demostrated that a prior history

of CUS resulted in the reduction of the
activational effects of previous exposure to
mild stressors on activity in a novel
environment.

This altered response was corrected by

chronic administration of different types of
antidepressants
A different CUS model (Armario et al. 1985)
 CUS (CV in the data) or
chronic inescapable tail-shock
(CS) induces behavioural
changes reminiscent of
depression

CS CV
 CUS induces anhedonia (Willner et al)

Is the same sucrose and

saccharin intake to
measure anhedonia?

(time 0 indicates the starting of antidepressant treatment. The

CUS was initiated several days before)

CUS reduced amphetamine and quinpirole-induced CPP, but enhanced dopamine release in the
NAC by electrical stimulation of the medial forebrain bundle, suggesting reduced presynaptic
inputs to the NAC rather than altered NAC response

A debate about the use of sucrose versus saccharin

Chronic social stress/defeat in rats and intracranial self-stimulation (ISS)
(Der-Avakian et al, Biol Psychiatry 76, 542, 2014)

Electrodes in the lateral hypothalamus

Immobility in the forced swim test did not parallel

changes in ISS
 Chronic social stress-induced cognitive bias

Rats are first trained to the positive tasks Discrimination training

and later to the negative one

Testing under Daily Social defeat 3 weeks Re-testing

ambiguous cues

Papciak et al, Behav Brain Res

256, 305, 2013
 Genetic differences in the response to the forced swim test and the response to
antidepressants: comparison of different inbred rat strains

WKY rats might be considered as a genetic model of depression resistant to

antidepressants
 Effort-related task

Salamone et al, Behav Processes (2016)

 Bipolar disorders and DSM-V
 Bipolar disordes are separated from depressive disorders. They are considered
in between schizophrenia spectrum (and other psychotic disorders) and depression.

 Classification:
 Bipolar I: requires at least a manic episode, but not a depressive episode.
 Bipolar II: manic or hypomanic episode together with major depression episode.
 Cyclotimic: hypomanic and depressive episodes over at least 2 years.
 Others

 Difference between manic and hypomanic episodes is intensity.

 Life-time prevalence of BP-I is 1% and that of BP-II is more difficult to establish,

but around 2% for both. No evident gender differences in prevalence, but some
differences in characteristics.

 Mean age onset about 18 years for BP-I and a bit latter for BP-II.
Hyperactivity
Euphoria, grandiosity
Irritability
Risk-taking
Suicide
 Neurobiological alterations in bipolar disorders

• Gray matter reduction particularly in prefrontal-ACC areas

• Reduction in sgACC similar in bipolar and major depression.

• Changes appear to precede the pathology and Lithium normalizes changes likely due
to its neurotrophic effects.

• Decreased fractional anisotropy (FA) of the uncinate fasciculus connecting OFC and
vmPFC/ACC with amygdala and hippocampus.

• Relationship with hippocampal volume is inconsistent and this migh be related to an

increase caused by lithium treatment.
Bipolar depression requires treatments completely different from other
types of depression

The most widely used treatments are Lithium,

anticonvulsants (carbamazepine and valproate) and
atypical antipsychotics . Other anticovulsants appear to
be less efficaceous and have not beeen aproved for BP
(lamotrigen, topiramate).

Mood stabilizers act on several intracelular pathways, but

the changes responsible for the therapeutic effects are
still unknown..

The main hypotheses about action of Li are related to the

inhibition of phosphoinositide-PKC pathways, the
reduction of the levels of myo-inositol (obtained from
glucose-6-P) or the inhibition of the activity of the
Glycogen synthase kinase-3 (GSK-3).
Factors contributing to cycling betwen mania and depression
(Young & Dulcis, Eur J Pharmacol 759, 151, 2015)
 Animal models for bipolar disorders
• Pharmacological models
– Psychostimulant-induced hyperactivity
• Environmental models
– Sleep-deprivation
– Stress-induced aggression
• Genetic models:
- Black Swiss mica (Swis x C57BL6/JN)
- Mice over-expresing GKS3.
- CLOCK (circadian rhythm gene) mutation encoding dominant negative protein
- D-box binding protein (transcription factor involved in circadian rhythms) KO
- Mutant form of mitochondrial DNA polymerase  in brain that affects mitochondrial function

Test to validate putative animal models

Prepulse inhibition

Hyperactivity in novel environments

Agressive behavior

Operant tasks evaluating sensitivity to reward or punishment or propensity to

risky behavior