US EPA

3236058 

Journal Article 

Distribution of NO-induced cGMP-like immunoreactive neurones in the abdominal nervous system of the crayfish, Procambarus clarkii 

Aonuma, H 

2002 

1 

Zoological Science
ISSN: 0289-0003 

19 

9 

969-979 

WOS:000181086100001 

Nitric oxide (NO) acts as a signalling molecule by
activating soluble guanylate cyclase and causing accumulation of the second messenger cyclic
guanosine 3',5'-monophosphate (cGMP) in target cells. In order to detect the presence of NO-
cGMP signalling pathway in the crayfish abdominal nervous system, accumulation of NO-induced cGMP
was investigated by anti-cGMP immunochemistry. Some preparations were incubated in a high-K+
saline containing an inhibitor of cGMP-degrading phosphodiesterase, 3-isobutyl-1-methyxanthine
(IBMX), to activate NO generating neurones, which could release NO in the ganglion, and then
immunohistochemistry using an anti-cGMP antibody was performed. The other preparations were
incubated in NO donor, sodium nitroprusside (SNIP) saline containing IBMX before anti-cGMP
immunohistochemistry was performed. The distribution of cGMP-like immunoreactive neurones in
high-K+ treated preparations was similar to that of cGMP-like immunoreactive neurones in NO donor
treated preparations. About 70-80 cell bodies and many neuronal branches in the neuropilar area
of the ganglion were stained, although no neurones showed immunoreactivity unless preparations
were activated by either high-K+ or the NO donor. Some of them were identical neurones, and they
were intersegmental ascending interneurones and motor neurones. Sensory afferents that innervates
hind gut showed strong cGMP-like immunoreactivity, although no mechanosensory afferents showed
any immunoreactivity. These results strongly suggest the presence of an NO-cGMP signalling
pathway that regulates neuronal events in the abdominal nervous system of the crayfish. 

crustacean; cGMP; high-K+; NO donor; immunohistochemistry