SCREENING MODELS IN PHARMACOLOGY DETAILED VIDEO ON EXPERIMENTATION MODELS.OBESITY DIABETES ALZHEIMER PARKINSON SCHIZPHRENIA ULCER PCOS CARDIOVASCULAR DISEASE ENDOCRINE DISEASE NEUROLOGICAL DISEASE PULMONARY DISEASE RENAL DISEASE HEPATOLOGICAL DISEAS SAFETY PHARMAOCLOGICAL STUDIES TOXICOLOGICAL STUDIES

1. EXPERIMENTAL
SCREENING MODELS
FOR VARIOUS DISEASES
BY PAYAAM VOHRA

2. Humanized
Organ on a chip
Transgenic
Knockin and knockout- db/db, BALB,ob/ob, SHR
Cell lines
Cell cultures

5. PHYSICAL
CHEMICAL
MECHANICAL
ACOUSTIC
VIBRATIONAL
ELECTRICAL
BIOLOGICAL

6. Arrhythmia type Experimental model
Wolff Parkinson white syndrome Transgenic PRKAG2 model
Atrial Flutter Atrial flutter induced by Ach and rapid pacing.
Atrial flutter by aconite.
Canine right Atrial occlusion injury model.
Atrial Fibrillation Atrial fibrillation in the isolated langendorff perfused
heart.
Canine model of chronic Atrial fibrillation.
PACAP-27 induced Atrial fibrillation.
Ventricular Fibrillation Ventricular fibrillation electrical threshold.
Canine model of two stage ligation.
Ventricular arrhythmia during exercise by ischemia.
6

7. DRUG ABUSE

8. In-Vivo METHODS
Chemically induced arrhythmia
Electrically induced arrhythmia
Mechanically induced arrhythmia
Exercise Induced arrhythmia
8

9. CHEMICALLY INDUCED ARRHTHMIA
A large number of chemical agents alone or in combination are capable of inducing arrhythmia. These include
1 Aconitine antagonism in rats
2 Digoxin induced arrthymia in guinea pigs
3 Strophanthin or ouabain induced arrhythmia in dog
4 Adrenaline induced arrhythmia
Coronary artery occlusion/reperfusion arrhythmia
Arrhythmias induced directly by ischemia and reperfusion
Coronary artery ligation in anesthetized dog results in
↑ in HR
↑in heart contractility
↑ in BP
Ventricular arrhythmias
MECHANICALLY INDUCED ARRHYTHMIA
ELECTRICALLY INDUCED ARHYTHMIAS
NERVE BLOOD FLOW
CONDUCTION VELOCITY
D-DIMER
CRP
9

12. ALZHEIMER

13. MODELS
SPONTANEOUS
CHEMICALLY
DRUG
INDUCED
TRANSGENIC
HYPOXIA
INDUCED
DENERVATION
BRAIN INJURY
THIAMINE
DEFICIENCY
HIGH FAT

14. DEPRESSION

15. STROKE

16. COLLAGENASE
BCAO MCAO
GLOBAL AND FOCAL
EMBOLIC
ENDOTHELIN
MICROBALLON INFLATION MODEL
FOOT-FAULT TEST MOTOR FUNCTION, LIMB COORDINATION
CORNER TEST SENSORIMOTOR ASYMMETRY
ADHESIVE REMOVAL TEST SENSORIMOTOR DYSFUNCTION, MOTOR ASYMMETRY

17. In vitro Models
ADD A FOOTER
17
In vitro models in depression are generally used as
preliminary high throughput screening for anti
depressant and are usually receptor binding studies
1. Inhibition of Dopamine/Serotonin uptake in rat striatal synaptosomes
2. Binding to monoamine transporters
3. Measurement of β-adrenoreceptor stimulated
4. Adenylate cyclase
5. Monoamine oxidase inhibition: Inhibition of type A and type B
monoamine oxidase activities in rat brain synaptosomes
6. Serotonin Binding Affinity
7. NMDA receptor Antagonism

18. Learned Helplessness
Model
ADD A FOOTER
18
• Following one or more sessions of inescapable shock, rats
have been shown to develop persistent changes including
weight loss, alterations in sleep patterns and HPA axis
activity and loss of spine synapses in hippocampal regions
• In mice, the learned helplessness (LH) syndrome appears to
be short-lived (2–3 days), and several mutant lines of mice
have been phenotyped on the LH assay, with results largely
compatible with their corresponding FST data.
• Like the FST or TST, both mice and rats display a
considerable degree of interstrain variation, and escape
deficits are reversed by a variety of antidepressants
Following an uncontrollable and inescapable
stress such as exposure to inescapable electric
shocks, animals develop a state of
“helplessness”

19. Chronic Mild Stress Model
19
• Chronic mild stress (CMS), better described as CUS, paradigms involve
the application of varied intermittent physical stresses applied over a
relatively prolonged time period (between 1 and 7 weeks
• Basically this model consist of various stressor applied throughout the
weeks randomly to impart stress on mice which gradually develops
anhedonic behaviour over the course of time
• CUS has also been shown to result in a number of other “emotional”
changes that are difficult to objectively quantify, such as grooming
deficits and changes in aggressive and sexual behaviour
• Aside from being a tool to study the physiological consequences of
chronic stress, CUS has been applied recently to phenotype mouse
mutants, study gender differences in stress responses, and validate
novel antidepressants
While acute stress paradigms are used broadly for their
ease, automation, and rapid phenotyping abilities, they
offer singular readouts that often cannot be
unambiguously interpreted

20. Genetic Models
Forward Genetics
20
There are two main approaches for genetic models of depression
Reverse Genetics
It is and unbiased approach in which large number of
random mutations are generated in the animal using
simple mutagenic techniques
This followed by breeding and screening for
individuals with the desired aberrant phenotype
After the generation of a mutant mouse line with the
desired phenotype—in this case, depressive-like
behaviour—the responsible gene can be identified.
The reverse genetic approach is used more commonly in scientific
practice
and involves genetic manipulations that result in either loss- or
gain of-function mutants.
“Knockout mice” are the most well-known examples, in which a
specific target gene is disrupted, resulting in a loss-of function
mutant.
However, loss of function can be achieved using other tools, such
as insertion of transgenes that produce an antisense mRNA of the
target gene or of short hairpin RNAs directed against the
gene of interest
For example, mouse lines expressing Cre recombinase, which is a
tyrosine recombinase enzyme derived from the P1 bacteriophage,
selectively in neurons of a specific neurotransmitter

21. ADD A FOOTER 21
Ontogenetic Mouse Model

22. Maternal Separation
22
• In particular, maternal separation was proposed to represent an important
animal model for investigation of the pathophysiology and treatment of
major depression
• mong the paradigms used to study early adverse life events, long maternal
separation in rodents mimics early life neglect/loss of parents in humans, and
has been presented as one of the most potent natural stressors during
development
• Maternal separation was developed to examine the consequences of early
adverse experiences on behaviour and neurobiology, and this model has been
described as a model of vulnerability to drug dependence, anxiety, stress-
induced illness, and depression
• The mice are separated from their mother at the time of birth for
unpredictable amount of time
• These mice when reach adult stage show hallmark signs of anhedonia and
depression
Early adverse life experiences represent one of the major
risk factors for the development of mental disorders such as
major depression.

23. Sleep Deprivation Model
23
• Increased levels of messenger RNA for interleukin-1b
(a pro-inflammatory cytokine) and for cortisol have been
shown in rodents after sleep deprivation
• The procedure of this study consisted of handling the animals
gently to prevent them from sleeping
• Furthermore, 72 hours of sleep deprivation in mice was induced
using the platform method, which is accomplished by placing the
animal on a platform submerged in water so that, when the
animal falls asleep, it falls into the water and must then climb
back onto the platform, thus forcing it to stay awake
• This study showed that after 72 hours of sleep deprivation,
there was an increase of oxidative stress in the
hippocampus
• Other sleep deprivation technique include the Flower Pot
Sleep has important homeostatic functions, and sleep
deprivation is a stressor that has consequences for the
brain as well as for many body systems.

24. Olfactory Bulbectomy (OBX)
Model
24
• OBX cause permanent changes in various behavioural parameters like
alterations in regular sleep pattern dysregulated cognitive function bizarre
sexual pattern, chronic stress hyperactivity
• The OBX model has been successfully detecting antidepressant drugs for
decades now with the most reproducible effect, meaning the hyperactivity in
the open field test
• This effect is also the confirmation test of successful surgery when the testing
drug is meant to affect a different parameter of the OBX rats
• Many effects of the bulbectomy are reversible with the administration of
antidepressant drugs and fail to return to baseline when given a drug lacking
antidepressant activity
• Such effects include decrease in passive avoidance learning deviating
leukocyte differential count and the microglial neuroinflammatory response
Bilateral olfactory bulbectomy (OB) results in endocrine,
behavioral, immune system, and neurotransmitter
changes that mimic many of the symptoms seen in
human patients with major depression.

25. Chemical Models Of Depression
Chronic Corticosterone
25
These models are generally based on pathophysiology of depression and unlike physical models employ one of
the mechanism of the developing depression, generally used in mechanistic studies of novel antidepressants
Reserpine Induced
This model is based on the Hypothalamus –
Pituitary – Adrenal Axis hyperactivity of
depepression
Chronic administration of corticosterone sends
the HPA axis on a hyper drive and is unable to
cope up even in mild stress condition
Gradually these mice through even slightest of
stressors or stimuli for short period can develop
depression
Reserpine was earlier widely used for treating
hypertension and schizophrenia
Observation show depressive like behaviour in
rodent thus reserpine models are then develop
Reserpine causes depletion of monoamine in
brain, a classic pathology of depression
Depressive symptoms are however accompanied
by other symptoms like hypothermia and ptosis

30. ULCER

31. SCREENINGMETHODS
IN VIVO METHODS
Pylorus ligation in rat
Isolated rat stomach
Stress ulcer models
Restraint- induced ulcer
cold water immersion induced ulcer
Histamine -induced ulcer
Ethanol -induced ulcer
NSAIDs- induced ulcer
Acetic acid- induced gastric ulcer 31
IN VITRO METHODS
[ 125I] Gastrin Binding Assay
Tiotidine Binding assay
H+/ K+ - ATPase inhibition Assay

32. OBESITY

33. Methods to induce
experimental obesity
1. Food induced obesity
obesity can be induced in rats by offering a diet containing corn oil and condensed milk
Male SD rats housed in controlled conditions
At the age of 6 months rats, divided into 2 groups, ordinary Purina Rodent Chow is fed to G1, the
the G2 is fed with Purina Rodent Chow, corn oil and condensed milk
Body weight and food intakes measured, sacrificed after 3 months
Adipose tissue cell size, lipid content, hormones, plasma lipids are determined.

34. 2. Hypothalamic obesity
Hyperphagia in rats induced after hypothalamic lesions
Female SD rats, fed HFD for 5-9 days, fasted over night, anesthetized with 35mg/kg
pentobarbital sodium+1mg atropine methyl nitrate
Bilateral knife cuts or electrolytic lesions sterotaxically positioned in the hypothalamus
Histological verification of placement of knife ciuts is made in brain fixed in 10% buffered
formalin and embeded in paraffin.
serial sections of hypothalamus are examined histologically

35. 3. Goldthioglucose- induced
obesity
i.p or i.m injection of goldthioglucose induces obesity in mice
Related to destruction of hypothalamic and extra thalamic areas of brain
Swiss albino mice of either sex, fed with commercial mouse chow
At the age of 6 months, single i.p injection of 30-40 mg/kg goldthioglucose
Food intake and body weight determined for 3 months.

36. 4. Monosodium glutamate-
induced obesity
Adiposity induced in mice by repeated s.c injections of MSG
Male charles river mice, treated immediately after birth with daily s.c MSG for 5
days
Animals weaned at 3 weeks age, housed under controlled temp.
Food consumption and body weight is measured at the regular intervals

37. Assays of anti-obesity activity
1. Anorectic activity (food consumption in rats)
Food intake and body weight measured in acute and semichronic experiments in normal or obese rats
respectively
Female zucker rats(250-350g), maintained under controlled conditions
Rats fed with special dishes along with test compound or treated by i.p for 7 days
Mazindol, 3mg/kg, a standard
2.APoE model

38. binding in brown adipose
tissue)
Brown adipose tissue, a major site for non-shivering thermogenesis in rodents
Obese male fatty Zucker rats(age; 13 weks, weight; 450g), receive multiple doses of test
compound in the drinking or tap water for 21 days
Food intake measured everyday, and bodyweight everyother day
Rats sacrificed, brown adipose tissue minced, diluted with 250mM sucrose and
homogenized
supernatant collected, centrifuged, BSA 0.2% added
After centrifugation, pellet suspended in albumin free-sucrose buffer
Binding of GDP to mitochondria is determined by incubating mitochondria in basic
medium containing 100mM sucrose, 20mM TES, 1mM EDTA, 1mM choline chloride,
2μM rotenone, and 10 μM 3H rotenone.

39. 3. Uncoupling protein and GLUT4 in brown
adipose tissue
Uncoupling proteins, family of inner mitochondrial membrane membrane transporters
dissipate the proton gradient
UCP1 expressed exclusively in brown adipocytes
Male fatty rats, 10 weeks age, given s.c injection of test compound
Sacrificed after 14 weeks treatment , brown and white adipose tissue removed
Northern and western blotting is performed , the total RNA and GLUT4 proteins
determined

40. 4. Resting metabolic rate
RMR, influenced by various drugs both in normal and obese animals
Female yellow KK mice and female C57B1 mice, age 12 weeks, housed under controlled conditions,
fed with commercial powdered chow and tap water
Test compound given i.m for 2 weeks
Daily food intake and eight is measured
RMR estimated by means of closed-circuit metabolic system, consists of chamber, circulating pumps,
desciccant and CO2absorbant canisters

41. SCREENING OF
SYMPATHOMIMETICS AND
SYMPATHOLYTICS

42. CARDIOVASCULAR ANALYSIS IN VIVO
 Α- AND Β-ADRENORECEPTORS IN THE MOUSE IRIS
 Α2-ADRENORECEPTOR BLOCKADE MEASURED IN VIVO BY
CLONIDINE-INDUCED SLEEP IN CHICKS
 ACTIVITY AT Β1- AND Β2-ADRENORECEPTORS IN THE RAT
 Β1- AND Β2-SYMPATHOLYTIC ACTIVITY IN DOGS
 INTRINSIC Β-SYMPATHOMIMETIC ACTIVITY IN RESERPINE-
PRETREATED DOGS

43. SYMPATHOMIMETIC DRUGS MAY RESULT IN
ANY ON FOLLOWING EVENTS LIKE
i. Mydriasis in the eye
ii. Enhanced stroke volume
iii. Acceleration of the heart rate
iv. Dialation of the coronary arteries
v. Constriction of the pulmonery vessels
vi. Relaxation of bronchial muscles
vii. Inhibition of gastric secretion
viii. Constriction of gastrointestinal sphincters
ix. Stimulation of uterus and
x. constriction of spleen capsule

44. INFLAMMATORY

45. ANTIPROLIFERATIVE ASSAY (I)
•Seed the cells in 24-well tissue culture plates at a density of 104
cells/cm2. That means 1 ml/well at a concentration of 2x104 cell/ml
(one well is considered 2 cm2 ).
•Prepare 3 wells/experimental point. For each experiment, blank and
solvent controls must be assayed along with at least 3 doses for
the test compound.
•Incubate the cells at 37o C in 9% CO2-air for a 24 h-recovery period.
•Treat the test cells with serial concentrations of drug/test
compound in an appropriate vehicle, in triplicate. The vehicle
should not exceed 1% of the tissue culture medium.
45

46. Carrageenan-Induced Paw Edema
DEXTRAN INDUCED
PLETHYSMOMETRY
HISTAMINE
BRADYKININ
Pleurisy model
Ctoron oil
Cotton Pellet Granuloma Test
Granuloma Pouch tes
Formalin induced

47. CNS DRUGS

48. Screening models for CNS stimulant/depressant drugs
In-vivo methods
Run away Test or Y Maze Test
Actophotometer
Open Field Test
Hole Board Test
Elevated Plus Maze
Forced Swim Test
Light Dark Test
Barbiturate induced sleeping time
Motor coordination
Tail Suspension test

49. Run away Test or Y Maze Test
This test is used to study the effect of a
drug on spontaneous activity and motor
coordination. Swiss albino rats of either
sex were selected. The mice were placed
individually in a symmetrical Y–shaped
runway (33 cm x 38 cm x 13 cm) for 3
min and the number of the maze wall 4
ft (an' entry’) were counted

50. Actophotometer
The locomotor activity can be easily
studied by using Actophotometer. Swiss
Albino mice of either sex (20 – 25g )
were randomly divided into three groups
of six animals. The rats were placed
individually inside the chamber of
actophotometer for 10 min and basal
activity score was noted. The animals
were treated with drug and after 30 min of
mice are placed again in actophotometer
for 10 min and the activity was
monitored. Percentage increase(in case of
CNS stimulant)/ decrease(in case of CNS
depressant) in activities were calculated.

51. Open Field Test
Mice were carried to the test room in their home cages and were
handled by the base of their tails at all times. Mice were placed into the
center or one of the four corners of the open field and allowed to
explore the apparatus for 5 minutes. After the 5 minute test, mice were
returned in their home cages and the open field was cleaned with 70 %
ethyl alcohol and permitted to dry between tests.
The Open Field Test provides simultaneous measures of locomotion,
exploration and anxiety.

52. Hole Board Test
Rats were transported from housing room to testing room inside their
home-cages to minimize transfer effect. To avoid possible visual
and/or olfactive influences, animals were allowed to acclimate for 30
minutes far from observational apparatus.
Each subject, experimentally naïve at test beginning, was placed in
the arena centre and allowed to freely explore for 10 min and this
gives the idea about CNS stimulant/depressant activity of drugs.
Experiments were recorded through a digital video camera and video
files stored in a personal Computer.

53. Elevated Plus Maze
This test has been widely validated to measure anxiety in
rodents.
This apparatus was made of Plexiglas and consisted of two
open arms (30cm × 5cm) and two closed arms (30cm ×
5cm) with 25cm walls. The arms extended from a central
platform (5cm ×5cm).
The maze was elevated 38.5cm from the room floor. Albino
rats of either sex (150 - 200 g) were randomly divided into
three groups of six animals.
Each animal was placed at the center of the maze, facing
one of the enclosed arms. Number of entries and the time
spent in enclosed and open arms was recorded for 5 min
test.
Entry into an arm was defined as the animal placing all four
paws onto the arm. All tests were taped by a video camera.

54. Forced Swim Test
Albino rats of either sex (150 - 200 g) were selected. Rats were placed
individually in a transparent glass cylinder (12 cm in diameter, height
25 cm), which was filled with water to a height of 15 cm. Two swim
sessions were conducted. An initial 15-min pre-test followed 24 hr
later by a 6 min test.
In the pre test session, the mice which have not yet treated were
forced to swim in a glass cylinder for 15 min. In the second session,
each mouse received a respective dose of sample 1 hour prior to test,
and placed in the cylinders again for 6 min.
The following behaviors were recorded during the last 4 min.
1. Immobility: floating in water without swimming.
2. Swimming: active movements of extremities and circling in the
container.
3. Climbing: active movements of forelimbs on the container wall

55. Light Dark Test
The apparatus consists of a plexiglass box with two compartments (20cm ×
20cm each), one of which illuminated compartment, facing one of the dark
areas.
The time spent in illuminated and dark places, as well as the number of
entries in each space, was recorded for 5 min
And this gives us an idea about the CNS stimulant/depressant property of
the drug.
White rats-black background
Black rats-White background

56. Motor coordination
Muscle coordination was determined
via the use of Ugo Basile Rota rod bar.
Swiss albino mice were placed on rota
rod prior to treatment and at 0.5, 1, 2,
3, 4 and 5 hrs after treatment. Any
mouse that fell off before the cut off
time 2 min were excluded from the
experiment

57. Tail Suspension test
For the test, mice were suspended
on the edge of a shelf, 58 cm above
the ground with adhesive tape
placed approximately 1 cm from the
tip of the tail. The duration of
immobility was recorded for a period
of 5 min after the drug treatment.

58. Anxiety models
 In vitro assay for GABAergic compounds:
 [3H]-GABA receptor binding
 GABAA receptor binding
 GABAB receptor binding
 Benzodiazepine receptor: [3H]-flunitrazepam binding assay
 Serotonin receptor binding
 General considerations
 Serotonin (5HT1A) receptor: binding of
 [3H]-8-hydroxy-2-(di-n-propylamino)-tetralin ([3H]-DPAT) .
 Serotonin (5HT1B) receptors in brain: binding of
 [3H]5-hydroxytryptamine ([3H]5HT)
 5HT3 receptor in rat entorhinal cortex membranes:
 binding of [3H]GR 65630
 Histamine H3 receptor binding in brain
In vitro models
58

59. Effect on behaviour
Methods based on unconditioned
(spontaneous) response
Exploratory activity
Elevated plus-maze
light-dark (two compartment box)
open field, closed field, etc.
Social behavior
social interaction
maternal separation, ultrasonic distress
calls
Predator
mouse defense test battery
human threat (primates)
predator’s call, odor associated
avoidance response
Methods based on unconditioned
(spontaneous) response
 Conflict models
 Vogel punished
drinking
 Geller-Seifter
conflict
 marmoset, pigeon
conflict models
 Other
 four plate test
 active/passive
avoidance learning
 conditioned
ultrasonic
vocalization
59

60. Anticonvulsant activity
Pentylenetetrazole (Metrazol) induced convulsions
Strychnine-induced convulsions
Picrotoxin-induced convulsions
Isoniazid-induced convulsions
Yohimbine-induced convulsions
PURPOSE AND RATIONALE
This assay has been used primarily to evaluate antiepileptic drugs. However, it
has been shown that most anxiolytic agents are also able to prevent or
antagonize chemical-induced convulsions
60

61. Elevated plus-maze
 Purpose The test has been proposed for selective identification of anxiolytic
and anxiogenic drugs
 Source of anxiety: open space, height, new environment
 Procedure: The rats (200–250 g body weight) are housed in pairs for 10 days
prior to testing in the apparatus Groups consist of 6 rats for each dose. Thirty
min after i.p. administration of the test drug or the standard, the rat is placed
in the center of the maze, facing one of the enclosed arms.
 Parameters measured:Time spent in the open arms entries into the open
arms time spent in the closed arms entries into the closed arms
total entries central time
61

62. Dimensions:
The plus-maze consists of two open arms, 50 × 10 × 40 cm,
and two enclosed arms, 50 × 10 × 40 cm,
50cm
62

63. 2.IN VIVO MODELS:
 Haffner’s Tail Clip Method
 Hot - plate test
 Electrical stimulation of the tail
 Grid - shock test
 Formalin test in rats
 Chemotherapy-Induced Pain
 Spinal Cord Injury
 Radiant heat method
 Tail immersion test

64. Ultrasonic distress calls
Source of anxiety: maternal separation
Parameters measured: time spent with ultrasonic
vocalization
total number of ultrasonic
vocalization
Anxiolytic effect: statistically significant
increase in either parameters
measured
;
64

65. Light-dark model
 Source of anxiety: light, novelty,
 Procedure
 An electronic system using four sets of photocells across the partition automatically counts movements through the
partition and clocks the time spent in the light and dark compartments. Naive male mice or rats are placed into the
cage. The animals are treated 30 min before the experiment with the test drugs or the vehicle intraperitoneally and
are then observed for 10 min Groups of 6–8 animals are used for each dose
 Parameters measured:
 Time spent in both area(horizontal, vertical activity) movement time in both area number of transitions
Anxiolytic effect:
 Statistically significant increasee in light movement time or number of transition
65

66. Social interaction
 PURPOSE AND RATIONAL
In an unfamiliar and brightly lit environment, the normal social interaction of rats (e.g. sniffing, nipping, grooming) is
suppressed. Anxiolytics counteract this
Suppression
 SOURCE OF ANXIETY:
presence of an unfamiliar social partner
PROCEDURE
Male Sprague-Dawley rats (225–275 g body weight) are housed in groups of 5 animals The apparatus used for the
detection of changes in social behaviour and exploratory behaviour consists of a Perspex open-topped box (51 × 51 cm
and 20 cm high) with 17 × 17 cm marked areas on the floor. One hour prior to the test,
two naive rats from separate housing cages are treated with the test compound orally. They are placed into the box (with
60 W bright illumination 17 cm above) and their behaviour is observed over a 10-min period by remote video
recording..
66

67. Purpose In the staircase paradigm, step-climbing
is purported to reflect exploratory or locomotor activity,
while rearing behaviour is an index of anxiety state.
The number of rearings and steps climbed are recorded
in a 5 min period.
Parameter measured The number of steps climbed and the number of rears are counted over a 3-
min period
Anxiolytic effect The number of rearings and steps climbed are recorded in a 5 min period.
The dissociation of these parameters
is considered to be characteristic for anxiolytic drugs.
The test was modified for rapid screening of anxiolytic
activity in mice
Staircase test
67

68. TOXICOLOGICAL
Teratogenicity/Reproductive Carcinogenicity Mutagenecity
Genotoxicity
SOMATIC
GERM
Female
Male

69. Organ anatomy
Hormone
prenatal developmen
embryogeniss
organogenesis
M I C R O N U C L E U S
D R O S O P H I L I A
C H R M O S O M A L A B B E R A T I O N S
A M E S
S E X L I N K E D A S S A Y S

70. Marble burying
 Source of anxiety: Presence of an unfamiliar
objects (potential source
of danger)
 Parameters measured: Number of buried marbles
 Anxiolytic effect: Statistically significant decrease in the number of buried
marbles
70

71. Stress-induced hyperthermia
(Handling order)
 Source of anxiety: Anticipatory anxiety,
handling, new environment
 Parameters measured: Core temperature in the
first three and last three
animals in each group
of 15 mice
 Anxiolytic effect: Body temperature of the last
three animals are not significantly
different from the first three
71

72. Vogel lick-conflict
Source of anxiety: Stressful situation
(48 h water depr.)
conflict between thirst
and punishment after drinking
Parameters measured: Number of accepted punishment (electric shock)
Anxiolytic effect: Statistically significant increase in the accepted shocks
72

73. mCPP-induced anxiety
 Source of anxiety: Chemically induced anxiety
 Parameters measured: Time spent in both side
(horizontal, vertical activity)
frequency of motion
number of transition
 Anxiolytic effect: Statistically significant
increase parameters
measured in the lit compartment or
in number of transition
73
Methylchloro phenoxy propionic acid

74. Geller conflict paradigm
PURPOSE AND RATIONALE
Experimentally induced conflict by punishing food rewarded
behaviour has been used to differentiate between various psychoactive drugs by Geller and
Seifter
EVALUATION
The total number of lever presses during the conflict periods (CRF) and the non-conflict
periods (VI) are counted
An increase of lever presses in the conflict period is regarded as indication of an anti-anxiety
effect,
a decrease of lever presses in the non-conflict period
as an indication for a sedative effect
74

75. CENTRALANALGESIC
ACTIVITY
1. IN VITRO MODELS:
* 3H-Naloxone binding assay
* 3H-Dihydromorphine binding to 𝜇 opiate
receptors in rat brain
* Receptor binding of nociceptin
* Bioassays for nociception
* Receptor binding of cannabinoids
* Vanilloid receptor binding