SEQUENCE AND REGULATION OF EUROPEAN EEL PROLACTIN MESSENGER-RNA

cDNA clones encoding the European eel (Anguilla anguilla L.) prolactin were isolated from a pituitary cDNA library constructed in gamma gt10 , using a rainbow trout Prl cDNA fragment as a probe. Four different i nserts were subcloned into the pGEM 3Z plasmid after PCR amplification . The 1082 bp-long nucleotide sequence revealed an open reading frame of 627 bp encoding a 24 amino acid-long signal peptide followed by a 1 85 amino acid-long mature protein. Comparison studies showed 60-70% ho mology with other known teleost fish prolactins and 30-45% with non-te leost fish, amphibian, reptilian, avian and mammalian prolactins. In s itu hybridization studies using labelled prolactin RNA probe showed a strong signal in the rostral pars distalis of the pituitary gland. We next examined the physiological regulation of this prolactin synthesis in vivo using Northern blot analysis and prolactin cDNA probe labelle d by random priming. The pituitary prolactin mRNA level was markedly d ecreased 3 weeks after transfer of eels from freshwater to sea water. Implants of thyroid hormones left for up to three weeks were ineffecti ve on prolactin mRNA. Estradiol administered as implant, alone or in c ombination with 500 mu g testosterone, was also unable to significantl y alter the pituitary mRNA level for prolactin in the freshwater silve r eels whatever the dose used (20-500 mu g) and whatever the duration of treatment (from 4 days to 10 weeks).