Gonadotropin-releasing hormone (GnRH) is produced by the hypothalamus and stimulates the synthesis and secretion of gonadotropin hormones. In addition, GnRH also stimulates the production and secretion of growth hormone (GH) in some fish species and in humans with certain clinical disorders. In the goldfish pituitary, GH secretion and gene expression is regulated by two endogenous forms of GnRH known as salmon GnRH and chicken GnRH-II. It is well established that protein kinase C (PKC) mediates GnRH-stimulated GH secretion in the goldfish pituitary. In contrast, the signal transduction of GnRH-induced GH gene expression has not been elucidated in any model system. In this study, we demonstrate, for the first time, the presence of novel and atypical PKC isoforms in the pituitary of a fish. Moreover, our results indicate that conventional PKC is present selectively in GH-producing cells. Treatment of primary cultures of dispersed goldfish pituitary cells with PKC activators (phorbol ester or diacylglycerol analogue) did not affect basal or GnRH-induced GH mRNA levels and two different inhibitors of PKC (Calphostin C and GF109203X) did not reduce the effects of GnRH on GH gene expression. Together these results suggest that, in contrast to secretion, conventional and novel PKCs are not involved in GnRH-stimulated increases in GH mRNA levels in the goldfish pituitary. Instead, PD98059 inhibited GnRH-induced GH gene expression, suggesting that the extracellular signal-regulated kinase (ERK) signaling pathway is involved. The results presented here provide novel insights into the functional-specificity of GnRH-induced signaling and the regulation of GH gene expression.