THE SOCIAL RECOGNITION TEST OF MEMORY IN THE RAT: A RELIABLE PRIMARY IN-VIVO COGNITION ASSAY FOR

GUIDING DRUG DISCOVERY/DEVELOPMENT DECISION-MAKING RELEVANT TO NEUROPSYCHIATRIC DISEASES

David Virley, Vincent Castagn, Elise Esneault and Anne-Marie Hernier

INTRODUCTION
The social recognition test in rats has become increasingly popular for the pharmaceutical industry as a tool to evaluate compounds for pro-cognitive activity. This memory test probes short-term recognition/working memory and provides a relatively high-throughput method to investigate
novel target mechanisms relevant to cognitive impairment associated with neuropsychiatric disorders such as Alzheimers Disease (AD), Schizophrenia and Parkinsons Disease (PD). This simple test can be configured in a number of formats to develop a rationale for the target
mechanism on temporally induced forgetting or reversing pharmacologically-induced impairments (e.g. scopolamine, MK-801) or aged-dependent deficits. Importantly the test uses spontaneous naturalistic behavior of an adult rat when exposed to a juvenile conspecific on two
occasions to access cognition, where the output measured (recognition index/ratio of investigation duration between the two sessions) involves an assessment of social exploration, strongly influenced by an olfactory component. As numerous mechanisms of diverse target classes have
been evaluated in this simple cognition assay, the level of pharmacological validation has greatly improved justifying its utility as a screening tool for drug discovery/development decision-making. Furthermore site of action and lesion studies have identified the frontal cortex as a key
anatomical substrate engaged in the social recognition memory task, providing a relatively simple method to examine novel target mechanisms on modulating frontal cognitive function with respect to social encounters. We provide evidence for a reliable deficit in spontaneous
recognition towards the familiar juvenile rat following a temporal delay (120 min) which is reversed by the cholinesterase inhibitor donepezil providing a useful benchmark for prosecuting and validating NCEs relevant to cognitive function, particularly with respect to AD and PD
indications. Furthermore robust pharmacologically-induced deficits are presented (e.g. scopolamine and MK-801) following a short delay (30 min) offering the opportunity of assessing the effects of novel targets on modulating cholinergic or glutamatergic tone, respectively. Finally we
present data on an aged-related social memory impairment following a short delay (30 min), relative to adult rats, providing a relevant paradigm to detect symptomatic and/or disease-modifying effects of novel mechanisms for neuropsychiatric disorders.

MATERIAL & METHODS Scopolamine-induced impairment

Donepezil improves memory in the temporal delay social recognition test
Male adult rats (Wistar-Han, 3-4 months of age, from Elevage Janvier, France) were assessed in a social in the social recognition test: Reversal by donepezil
recognition memory test to recall prior exposure to a conspecific juvenile (3-4 weeks of age). Adult rats were 1st contact (T0-5') 2nd contact (T120'-125') Recognition index 1st contact (T0-5') 2nd contact (T30'-35') Recognition index
housed individually and all testing was carried out in their home cage. An unfamiliar juvenile was introduced
and overall investigation (sniffing, grooming, licking and closely following behaviors) was recorded for a 5- 120
Same juvenile
1.50 120
Different juvenile
1.50 120 1.50

Investigation duration (s)

Investigation duration (s)

Investigation duration (s)
min period (contact 1 = C1). The juvenile rat was then removed to its home cage. After 120 min, the same

Recognition index
100 1.25 100 1.25 100 1.25

Recognition index
Recognition index
juvenile was reintroduced and overall investigation duration was again recorded during a second 5-min ** (b)
period (contact 2 = C2). Donepezil (1 - 3 mg/kg i.p.) or vehicle was administered i.p. to the adult rat, 80 ** (b) ** (b) 1.00 80 1.00 80 1.00
immediately after C1. A recognition index (=C2/C1) was also calculated. To control for non-specific effects, 60
** (b)
0.75 60 0.75 60 0.75
** (a) * (a)
a separate experiment was carried out in which a new different/unfamiliar juvenile was introduced to the *** (a)
*** (a) ** (c)
40 0.50 40 0.50 40 0.50
adult rats during C2. To evaluate the effect of a pharmacologicallyinduced social recognition impairment,
scopolamine (0.25 mg/kg i.p.) or MK-801 (0.05 and 0.1 mg/kg i.p.) was administered 30 min prior to C1 20 0.25 20 0.25 20 *** (a) 0.25

relative to vehicle i.p.. Following a 30-min delay adult rats were re-exposed to the juvenile conspecific (C2). 0 0.00 0 0.00 0 0.00
Vehicle Donepezil 1 mg/kg Donepezil 2 mg/kg Donepezil 3 mg/kg Vehicle Donepezil 1 mg/kg Donepezil 2 mg/kg Donepezil 3 mg/kg Vehicle + Vehicle Vehicle + Donepezil 2.5 mg/kg +
To test potential reversal, donepezil (2.5 mg/kg i.p.) was administered 60 min prior to scopolamine and Scopolamine 0.25 mg/kg Scopolamine 0.25 mg/kg

clozapine (0.3 mg/kg i.p.) was administered 60 min prior to MK-801. To establish an aged-dependent effect (a) compared with first contact: no indication = not significant; * = P < 0.05; ** = P < 0.01; *** = P < 0.001 (Investigation duration) (a) compared with first contact: no indication = not significant; *** = P < 0.001 (Investigation duration)
(b) compared with vehicle control: no indication = not significant; ** = P < 0.01 (Recognition index) (b) compared with vehicle control: no indication = not significant; ** = P < 0.01 (Recognition index)
on social recognition performance, male Wistar (Han) rats of 20 22 months of age were compared to adult (c) compared with scopolamine: no indication = not significant; ** = P < 0.01 (Recognition index)

rats using a 30-min temporal delay. All data were evaluated statistically using an ANOVA followed by
appropriate post-hoc tests.
MK-801-induced impairment
Aged rats perform poorly in the social recognition test
in the social recognition test: Reversal by clozapine
RESULTS 1st contact (T0-5') 2nd contact (T30'-35') Recognition index 1st contact (T0-5') 2nd contact (T30'-35') Recognition index
A temporal delay of 120 min between C1 and C2 induced a robust impairment in spontaneous recognition in adult rats 120 1.50 120 1.50

Investigation duration (s)

Investigation duration (s)

* (b)
towards the same/familiar juvenile rat. Importantly the cholinesterase inhibitor donepezil (1 - 3 mg/kg i.p.) improved

Recognition index
100 1.25 100 1.25

Recognition index
* (b)
social recognition memory performance of the adult rat using this temporal delay when administered immediately after C1
80 1.00 80 1.00
(Ps<0.05). This was not apparent when a different/unfamiliar juvenile was presented to the adult rat on C2 (Ps>0.05, * (c)
NS), suggesting that the reversal of the deficit by donepezil is a function of re-exposure to the same/familiar juvenile. 60 * (a) 0.75 60 0.75
*** (a)
Scopolamine (0.25 mg/kg i.p.) or MK-801 (0.05 mg/kg i.p.) administered 30 min prior to C1 induced a significant 40 0.50 40 * (a) 0.50
recognition deficit during C2 following a 30-min delay (Ps<0.05), suggesting that both cholinergic and glutamatergic
20 0.25 20 0.25
blockade disrupts social working memory. Furthermore donepezil (2.5 mg/kg i.p.) reversed the scopolamine-induced
impairment (P<0.01) while clozapine (0.3 mg/kg i.p.) tended to reverse the MK-801-induced impairment (P<0.05). Aged 0 0.00 0 0.00
Vehicle + Vehicle Vehicle + Clozapine 0.3 Vehicle + Clozapine 0.3
Adult Control Aged Control
rats (>20 months of age) demonstrated a significant social recognition memory deficit (P<0.05) following a short (30 min) MK-801 0.05 mg/kg + MK801
mg/kg 0.05 mg/kg
MK801 0.1
mg/kg
mg/kg + MK-
801 0.1 mg/kg
delay as compared to adult rats (3-4 months of age) confirming the natural occurring decline in social cognitive function as
(a) compared with first contact: * = P < 0.05; *** = P < 0.001 (Investigation duration) (a) compared with first contact: no indication = not significant; * = P < 0.05 (Investigation duration)
a function of age. (b) compared with vehicle control: * = P < 0.05 (Recognition index) (b) compared with adult control: * = P < 0.05 (Recognition index)
(c) compared with MK-801 alone: * = P < 0.05 (Recognition index)

CONCLUSION

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The social recognition test in the rat provides a relevant short-term working memory test to evaluate the pro-cognitive activity of numerous target mechanisms aligned to cognitive impairment
associated with neuropsychiatric disorders, such as Alzheimers Disease, Schizophrenia, Parkinsons disease. Importantly this test can be used in a number of configurations to evaluate the
effects of novel target mechanisms on natural forgetting cognitive processes i.e. consolidation, as well under the experimental conditions involving pharmacological manipulations such as
scopolamine and MK-801. Importantly the selection of the format of the test using pharmacologically-induced impairments can help provide mechanistic insight for NCEs related to cholinergic
or glutamatergic processes to support the subsequent progression to alternative tests probing different cognitive domains and anatomical substrates relevant to neuropsychiatric disorders. The
aged-dependent deficits in social recognition memory offers the potential for determining the symptomatic and/or disease-modifying properties of a NCE, providing a more disease-relevant
assessment for drug discovery/development decision-making.

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