Visualization of nitric oxide production in the earthworm ventral nerve cord

NADPH-diaphorase histochemistry selectively stains peripheral and central sensory structures of lumbricid earhworms

“…The presence and pattern of NADPH-d labelled neurons were shown in the CNS of leeches [13], which are closely related to earthworms, so the organization of the VNC ganglia seems to be similar, but the staining was restricted to perikarya and no characteristic labelling was described in the neuropile. In contrast to an earlier work focused on the investigation of the NADPH-d stained structures of earthworm CNS [12], this study presented not only an account of NADPH-d reactivity of several structures in earthworms' VNC ganglia but also the exact anatomical positions of some distinct structures.…”

“…The occurrence of putative NO-producing (NADPH-d positive) structures was partially described in the VNC of Lumbricus terrestris [12], however, without exact anatomical characterization.…”

Distribution of NO-Induced cGMP-Like Immunoreactive Neurones in the Abdominal Nervous System of the Crayfish, Procambarus clarkii

“…Crayfish abdominal ganglia were dissected out and they were preincubated in 1 mM 3-isobutyl-1-methyxanthine (IBMX; Sigma, St. Luis, MO) in the saline for 30 min at 4 ° C to block endogenous phosphodiesterase activity. Incubation in high-K + saline could depolarize membrane potential of all neurones (Oka et al ., 1994;Oka and Ogawa, 1996) including NO generating neurons, which release NO in the ganglion (Kitamura et al ., 2001). Preparations were then incubated in high-K + crayfish saline (NaCl: 5.36 mM, KCl: 205.3 mM, CaCl 2 : 13.54 mM, MgCl 2 : 2.62 mM, NaHCO 3 : 2.38 mM, HEPES 5 mM, pH 7.4) containing 1 mM IBMX for 15 min at 20 ° C to detect NO-related increase in cGMP.…”

“…High-K + saline activates NO generating neurones to generate endogenous NO in ganglion (Oka et al ., 1994;Oka and Ogawa, 1996;Kitamura et al ., 2001). Anti-cGMP immunohistochemistry in wholemount preparation revealed that about 75 cell bodies (76 ± 16, mean ± SD, n=6) and many neuronal branches were strongly stained in the terminal abdominal ganglion (Fig.…”

“…Preparations that are incubated in NO donors accumulate cGMP which are detectably stained by anti-cGMP immunohistochemistry. It has been demonstrated that high-K + saline depolarized the membrane potential (Oka et al ., 1994;Oka and Ogawa, 1996) and activated NO generating cells to release NO in a ganglion (Kitamura et al ., 2001), which in turn stimulated soluble guanylate cyclase in the target cells. In the present study, anti-cGMP immunohistochemistry is performed after stimulating soluble guanylate cyclase by incubating preparations in high-K + saline or NO donor saline to detect the target cells of NO in the abdominal ganglion of the crayfish.…”