Elsevier

Nitric Oxide

Volume 10, Issue 3, May 2004, Pages 130-140
Nitric Oxide

Neuronal nitric oxide synthase knock-out mice show impaired cognitive performance

https://doi.org/10.1016/j.niox.2004.03.007Get rights and content

Abstract

Nitric oxide (NO) plays a role in a series of neurobiological functions, underlying behavior and memory. The functional role of nNOS derived NO in cognitive functions, however, is elusive. We decided to study cognitive functions in the Morris water maze (MWM) and the multiple T-maze (MTM) in 3-month-old male nNOS-knock-out mice (nNOS KO). To study the influence of neurology and behavior, we performed tests in an observational battery, the rota-rod, the elevated plus maze (EPM), the open field (OF), and a social interaction test. In the memory and relearning task of the MWM, most nNOS KO failed whereas performing better in the MTM. nNOS KO displayed significantly increased frequency of grooming, center crossings, and entries into the center in the OF. The observational battery revealed significantly increased scores for touch–escape reaction, body position, locomotion, and pelvic- and tail-elevation together with reduced vocalization. In the EPM, the time spent in the closed arm and the grooming frequency were significantly increased whereas urination was absent. We conclude that nNOS KO show impaired spatial performance in the MWM and herewith confirm the role of nNOS in cognitive functions such as processing, maintenance, and recall of memory. It must be taken into account that the major behavioral findings of increased grooming and anxiety-related behaviors may have led to impaired function in the MWM. The fact that nNOS KO performed well in the MTM, reflecting a low stress situation points to the interpretation that nNOS inhibition affects cognitive functions under stressful conditions (MWM) only.

Section snippets

Animals

Protocols of experiments were approved by the Bundesministerium für Bildung, Wissenschaft und Kultur, Kommission für Tierversuchsangelegenheiten, Austria. All animals were housed in groups under controlled conditions of temperature, light, and humidity with food and water available ad libitum. Male mutant mice with targeted disruption of the nNOS gene (nNOS KO), showing >95% loss of NO production in the CNS [3] and wild-type (WT) animals C57BL/6J, which share >99.9% genetic similarity with the

NADPH-diaphorase enzyme histochemistry

WT animals exhibited in the caudate putamen and amygdaloid complex (particularly in the medial amygdala) an intense NADPH-d staining, which was virtually absent in nNOS KO animals and only spared in some blood vessels at the amygdaloid level. Similarly, NADPH-d staining was almost absent in nNOS KO in the hypothalamic supraoptic nucleus whereas WT mice showed an intense labelling. As expected, in nNOS KO mice NADPH-d staining was absent or greatly reduced at all remaining brain areas

Discussion

The major outcome of our study is the observation that genetic blockade of nNOS leads to impairment of cognitive functions (memory recall and relearning) under the stressful conditions of the MWM procedure.

Acknowledgements

We are highly indebted to the Red Bull Company, Salzburg, Austria, for generous support of the study. We kindly appreciate the contribution of the Verein “Unser Kind,” Verein zur Durchführung der wissenschaftlichen Forschung auf dem Gebiet der Neonatologie und Kinderintensivmedizin.

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