The distribution of growth hormone-releasing hormone-like peptides (GHRH-LP) in the central nervous system of the zebrafish was investigated by using immunohistochemical techniques with polyclonal antibodies. ELISAs showed that the antiserum raised against salmon (s)GHRH-LP recognized both zebrafish GHRH-LP1 and -2, whereas the antiserum raised against carp (c)GHRH-LP was more sensitive but detected only zebrafish GHRH-LP1. Neither antiserum detected the true GHRH. Large cells in the nucleus lateralis tuberis were immunoreactive with both antisera, which suggests that they contained zebrafish GHRH-LP1, but not excluding GHRH-LP2. Also, immunoreactive fibers, which putatively originated from these hypothalamic neurons, were present in the hypophysis; both antisera detected fibers, although only sGHRH-LP antiserum stained fibers in the neurointermediate lobe. These fibers may have a neuroendocrine role. Candidates for a role in feeding include several areas in which both antisera labeled cells and fibers, implying a strong reaction for GHRH-LP1 and possibly GHRH-LP2. These areas include the isthmus with cells in the secondary gustatory/visceral nucleus, which were also calretinin immunoreactive. Numerous GHRH-LP-immunoreactive fibers (also labeled by both antisera) probably originate from the gustatory/visceral nucleus to innervate the ventral area of the telencephalon, preglomerular nuclei, torus lateralis and hypothalamic diffuse nucleus, habenula, torus semicircularis, and dorsolateral funiculus of the spinal cord. Present results in the zebrafish brain suggest involvement of GHRH-LP in both neuroendocrine and feeding-associated nervous circuits. The present data on the location of the two GHRH-LPs are the first clue to the possible functions of these two hormones.
2009 Wiley-Liss, Inc.