Elsevier

Neuropharmacology

Volume 42, Issue 3, March 2002, Pages 414-420
Neuropharmacology

Neonatal nitric oxide synthase inhibition: social interaction deficits in adulthood and reversal by antipsychotic drugs

https://doi.org/10.1016/S0028-3908(01)00180-0Get rights and content

Abstract

Nitric oxide synthase (NOS) is thought to migrate improperly during development in the brains of schizophrenic patients. Also it is known that nitric oxide (NO) effects synaptogenesis during development of the CNS. Previously we have shown that neonatal treatment with a NOS inhibitor effects an animal's sensitivity to amphetamine and PCP. In the present study, neonatal rats were challenged with a NOS inhibitor (l-nitroarginine, 10 mg/kg, s.c.) daily on post-natal days (PD) three, four and five. l-Nitroarginine (l-NoArg) treated male rats at adulthood (PD56 and older) had a deficit in social interaction (SI) when placed in an environment with another foreign male rat and this deficit was reproducible on a weekly basis for at least five weeks. Haloperidol failed to significantly reverse this deficit before pronounced secondary effects on general behavior were seen at high doses. However, the atypical antipsychotics, clozapine and olanzapine, were able to significantly reverse this deficit at doses which did not effect baseline SI values. In a separate cohort of animals the effect of DOI was investigated, this was done to ascertain if there was a differential sensitivity of serotonergic pathways in this model. There was no difference in the behavioral score elicited from control or NoArg-treated rats. It is suggested that the SI deficits seen here may be more sensitive to atypical antipsychotics rather than haloperidol.

Introduction

The etiology of schizophrenia is poorly understood. One theory is that disturbances in the development of the brain may in part be responsible for the breakdown of normal neural control in later life (Weinberger, 1987, Bloom, 1993, Bogarts, 1993, Weinberger and Lipska, 1995). In order to explore this theory, one can investigate developmental mechanisms and study their consequences in adulthood.

The formation of the medial temporal lobe appears to be affected in those patients who subsequently exhibit schizophrenia. Of particular interest were studies showing that NADPH-d (nicotinamide-adenine dinucleotide phosphate-diaphorase) neurons migrate improperly in the prefrontal cortex, hippocampal formation and lateral temporal lobe of schizophrenic patients (Akbarian et al., 1993a, Akbarian et al., 1993b, Bloom, 1993). Therefore the developing brain of a person who later becomes schizophrenic may have an abnormal distribution of NADPH-d containing neurons, particularly in those brain areas intimately linked to the disease. NADPH-d is a nitric oxide synthase capable of generating nitric oxide (NO) (Hope et al., 1991, Dawson et al., 1991). NO is formed endogenously by the conversion of l-arginine to l-citrulline by the enzyme nitric oxide synthase (NOS) (Mayer et al., 1989). NOS is known to exist in at least three isoforms (neuronal NOS, endothelial NOS and inducible NOS) broadly corresponding to that present in neuronal, endothelial and macrophage cells (Snyder and Bredt, 1991). In the rat, NADPH-d neurons are present at birth and then increase in number markedly throughout the neocortex during the first post-natal week (Tomic et al., 1994, Yan et al., 1994), and reach adult levels and laminar distribution by the end of the second post-natal week. This is mirrored by analysis of NOS activity in the rat neocortex where the first nine days post-natal are the most active for neuronal NOS; adult levels of NOS activity are substantially less than neonatal levels (Ogilvie et al., 1995). NO has been hypothesized to influence the maturation of neurons and synaptogenesis during neuronal development (Gally et al., 1990, Hess et al., 1993, Ogilvie et al., 1995, Wu et al., 1996).

We have previously hypothesized that interfering with NO production during the very early postnatal period may reproduce some of the aspects of schizophrenia in animals (Black et al., 1999a, Black et al., 1999b, Simmonds et al., 2000). Neonatal rats treated with a NOS inhibitor exhibited amphetamine and PCP hypersensitivity as well as pre-pulse inhibition disruption in adulthood. In order to explore this model further we have examined social interaction (SI) behavior and serotonergic influences. Social interaction has been used previously to investigate cognition processes (Engelmann et al., 1995, Mondadori et al., 1996), however, we used this methodology to try to mimic the frequently seen negative schizophrenic symptom of social withdrawal. Also this behavioral method is not driven by a pharmacological stimulus such as amphetamine or PCP and therefore could be studied, as a pure effect not biased by drug action. 5HT2A receptors are known to be involved in schizophrenia (for review see Meltzer, 1999) therefore we decided to investigate a serotonergic component of this model by determining the sensitivity of rats to DOI (a 5HT2A/2C agonist). In this regards, the attenuation of DOI induced behaviors has been used as a test for antipsychotics (Wettstein et al., 1999). These data presented here are an attempt to dissect apart this neurodevelopmental model and assess its utility in drug discovery research. We have chosen to use a typical antipsychotic, haldol, and the atypical antipsychotics clozapine and olanzapine. It is generally recognized that atypical antipsychotics treat negative symptoms (e.g. social withdrawal) better than typical antipsychotics (Campbell et al., 1999).

Section snippets

Animals

Sprague–Dawley rats (Charles River, USA) were used throughout the study. A single female rat and her litter were housed individually. At weaning (day 22), male pups were separated and housed 4–5 to a cage in the same room. Female pups were not used in experimentation. Rats had access to food and water ad libitum with a 12-hour light/dark cycle. All testing was performed during the light (day) cycle. No assessment of the stage of estrous cycle was made at any time during the behavioral testing

Results

After PD56 (Week 1) the rats were examined for baseline SI. Animals treated with l-NoArg had a SI deficit when compared to phosphate-buffered saline treated (control) animals (Fig. 1). A deficit is defined as a decrease in interaction time of the adult with the juvenile animal compared to a control situation. Other than this deficit no other overt behavioral differences were seen between control and NoArg treated animals

During Week 2, animals were given a low dose of haloperidol, clozapine or

Discussion

Neonatal treatment of male rat pups with l-NoArg elicited a reproducible deficit in SI over a period of several weeks in adulthood. This interaction was more sensitive to the atypical antipsychotics (clozapine and olanzapine) than the typical antipsychotic (haloperidol). This deficit is not dependent on stimulation by exogenous psychoactive compounds and therefore offers investigators an animal model with which to potentially study schizophrenia free of psychostimulant bias. While these results

Acknowledgments

The authors wish to thank Frank Todd for his excellent technical assistance.

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