In vitro effects of mammalian leptin, neuropeptide-Y, beta-endorphin and galanin on transcript levels of thyrotropin beta and common alpha subunit mRNAs in the pituitary of bighead carp (aristichthys nobilis)

Abstract

Thyrotropin (thyroid stimulating hormone, TSH) is a member of the pituitary glycoprotein hormones, consisting of two dissimilar subunits, alpha and beta. The two subunits are produced by different genes and are regulated independently. We have previously cloned a TSHbeta cDNA from bighead carp pituitary and investigated its gene regulation. We report here the direct effects of mammalian TSH-releasing hormone (TRH), leptin, neuropeptide-Y (NPY), beta-endorphin and galanin on mRNA levels of both TSHbeta and alpha-subunits in the pituitary of bighead carp in vitro. The dispersed pituitary cells of bighead carp were incubated at 25 degrees C for 6 h with different doses of these factors. The relative mRNA levels of TSHbeta and alpha-subunits were estimated by traditional polymerase chain reaction (PCR) analysis and fluorescence real-time PCR analysis. The results revealed that mammalian TRH, leptin and beta-endorphin produced dose-dependent stimulatory effects on mRNA levels of both TSHbeta and alpha-subunits while thyroxine (T4) and mammalian galanin suppressed mRNA levels of both TSHbeta and alpha-subunits. NPY suppressed TSHbeta mRNA level, but stimulated alpha-subunit mRNA level. This study has demonstrated that mammalian TRH, leptin, NPY, beta-endorphin and galanin were active in modulating the steady-state mRNA levels of TSHbeta and alpha-subunits of bighead carp pituitary in vitro. The results suggest that endogenous TRH, leptin, NPY, beta-endorphin and galanin may modulate transcript levels of TSHbeta and alpha-subunits in pituitary of bighead carp.