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First published online February 6, 2004
Journal of Experimental Biology 207, 923-935 (2004)
Published by The Company of Biologists 2004
doi: 10.1242/jeb.00845
The early ontogeny of neuronal nitric oxide synthase systems in the zebrafish
1 Department of Pathology, Lund University, Sölvegatan 25, S-221 85
Lund, Sweden
2 Department of Anatomy and Neurobiology, Boston University Medical School,
715 Albany Street R-91, Boston, MA 02118-2394, USA
* Author for correspondence (e-mail: bo.holmqvist{at}pat.lu.se)
Accepted 17 December 2003
To examine a putative role for neuronal nitric oxide synthase (nNOS) in early vertebrate development we investigated nNOS mRNA expression and cGMP production during development of the zebrafish Danio rerio. The nNOS mRNA expression in the central nervous system (CNS) and periphery showed a distinct spatio–temporal pattern in developing zebrafish embryo and young larvae. nNOS mRNA expression was first detected at 19 h postfertilisation (h.p.f.), in a bilateral subpopulation of the embryonic ventrorostral cell cluster in the forebrain. The number of nNOS mRNA-expressing cells in the brain slowly increased, also appearing in the ventrocaudal cell cluster from about 26 h.p.f., and in the dorsorostral and hindbrain cell cluster and in the medulla at 30 h.p.f. A major increase in nNOS mRNA expression started at about 40 h.p.f., and by 55 h.p.f. the expression constituted cell populations in differentiated central nuclei and in association with the proliferation zones of the brain, and in the medulla and retina. In parts of the skin, nNOS mRNA expression started at 20 h.p.f. and ended at 55 h.p.f. Between 40 and 55 h.p.f., nNOS mRNA expression started in peripheral organs, forming distinct populations after hatching within or in the vicinity of the presumptive swim bladder, enteric ganglia, and along the alimentary tract and nephritic ducts. Expression of nNOS mRNA correlated with the neuronal differentiation pattern and with the timing and degree of cGMP production.
These studies indicate spatio–temporal actions by NO during embryogenesis in the formation of the central and peripheral nervous system, with possible involvement in processes such as neurogenesis, organogenesis and early physiology.
Key words: morphogenesis, zebrafish, Danio rerio, brain, retina, gut, intestine, hybridisation, in situ, development, regeneration, neuronal differentiation
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