Melanin-concentrating hormone reduces somatolactin release from cultured goldfish pituitary cells

Melanin-concentrating hormone (MCH)-containing neurons directly innervate the adenohypophysis in the teleost pituitary. We examined immunohistochemically the relationship between MCH-containing nerve fibres or endings and somatolactin (SL)-producing cells in the goldfish pituitary. Nerve fibres or endings with MCH-like immunoreactivity were identified in the neurohypophysis in close proximity to the adenohypophysial cells showing SL-like immunoreactivity. We also examined the effect of MCH on SL release from Cultured goldfish pituitary cells and SL synthesis using a cell immunoblot and a real-time PCR method. Treatment of individually dispersed pituitary cells with MCH 10(-7) M for 3 h decreased the area of SL-like immunoreactivity on immunoblots, and MCH-induced reductions in SL release were blocked by treatment with the mammalian MCH receptor (MCHR) antagonist, compound-30, at a concentration of 10(-5) M. Treatment with 10(-7) M MCH for 3 h did not affect sl-alpha and -beta (smtla and -b as given in the Zfin Database) mRNA expression levels. These led us to explore the signal transduction mechanism leading to the inhibition of SL release, for which we examined whether MCH-induced reductions in SL release are mediated by the G(i) or G(q) protein-coupled signalling pathway. The MCH-induced reductions in SL release were abolished by treatment with the G(i/o) protein inhibitors, NF023 (10(-5) M) or pertussis toxin (260 ng/ml), but not by the phospholipase C inhibitor, U-73122 (3 x 10(-6) M). These results indicate that MCH can potentially function as a hypothalamic factor suppressing SL release via the MCHR, and subsequently through the G(i) protein to inhibit the adenylate cyclase/cAMP/protein kinase A-signalling pathway in goldfish pituitary cells. journal of Endocrinology (2009) 203, 389-398