Early observations of nitric oxide synthase in the intact brain and regenerating tentacle of the snail, Helix pomatia
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1
Balaton Limnological Research Institute, HAS, Department of Experimental Zoology, Hungary
In the present study, the regeneration dynamics of the Helix tentacle which is an established model for neuronal as well as organ regeneration was investigated by NADPH-diaphorase (NADPH-d) reaction which is the histochemical marker for nitric oxide synthase (NOS). The NOS activity and expression, as well as its possible impact on cysteine and tyrosine nitrosylation were also followed in intact brain and tentacle. In a period of 15 weeks, the terminal part of the right tentacle of ten animals was cut by every odd week; meanwhile the left tentacle of these groups remained intact for control. Neurons of the tentacular ganglion, their projections in the tentacular nerve innervating the lateral neuropil and the globuli cells of the procerebrum, showed NADPH-d reactivity. After 1 week of tentacle extirpation a faint NADPH-d reactivity was still seen in the distal part of injured axons of the tentacular nerve. At 10 weeks of regeneration NADPH-d reactivity reappeared in the newly forming tentacular ganglion, and a similar distribution to the control was obtained by 13 weeks. NOS activity of the brain was found 1422±42 dpm 14C citrulline/mg/ml protein/min/5 animals (n=4), whereas it was 2212±50 dpm in the tentacular ganglia. A fivefold dilution of the extracts resulted in an approx. five times higher activity in the brain (6691±193 dpm; n=4). L-NAME substrate analogue inhibitor proved not to be effective. From total brain extract a 140-150 kDa protein was detected by an anti-universal NOS antibody that was also labeled by an anti-S-nitrosocysteine antibody. In addition, cysteine nitrosylation was also found in other 5 proteins of 180, 170, 80, 50, and 40 kDa mw, respectively, whereas tyrosine nitration was not present in the intact Helix brain. These preliminary results show the presence of the NO signaling pathway in the snail CNS and tentacle and also suggest a possible involvement of NO in the tentacle regeneration.
Supported by the OTKA PD75276 and K78224 grants.
Conference:
IBRO International Workshop 2010, Pécs, Hungary, 21 Jan - 23 Jan, 2010.
Presentation Type:
Poster Presentation
Topic:
Sensory and motor systems
Citation:
Serfözö
Z and
Elekes
K
(2010). Early observations of nitric oxide synthase in the intact brain and regenerating tentacle of the snail, Helix pomatia.
Front. Neurosci.
Conference Abstract:
IBRO International Workshop 2010.
doi: 10.3389/conf.fnins.2010.10.00267
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Received:
06 May 2010;
Published Online:
06 May 2010.
*
Correspondence:
Zoltán Serfözö, Balaton Limnological Research Institute, HAS, Department of Experimental Zoology, Tihany, Hungary, serfozo@tres.blki.hu