Cloning and Phylogenetic Relationship of Red Drum Somatolactin cDNA and Effects of Light on Pituitary Somatolactin mRNA Expression

Abstract

The nucleotide sequence for red drum somatolactin (SL) cDNA was determined and the expression of pituitary SL mRNA was examined in red drum kept under various light conditions. A full length of SL cDNA (1629 bp) was isolated and characterized from a red drum pituitary cDNA library. The SL cDNA has an open reading frame of 696 nucleotides which encodes a 24-amino-acid signal peptide and a 207-amino-acid mature peptide. Red drum SL shares 58–87% amino acid sequence identity and 56–85% nucleotide sequence identity with other teleost SLs. The characteristic seven cysteine residues and one N-glycosylation site of SL are well conserved in the red drum SL mature peptide. Phylogenetic analysis shows that red drum SL is closely related to seabream SL and is also closely related to lumpfish, flounder, halibut, and sole SLs, whereas SLs of Atlantic cod, chum salmon, rainbow trout, and eel are more distantly related to those of the more advanced teleosts. Two SL transcripts, designated as SL I at 1.8 kb and SL II at 1.3 kb, are expressed in red drum pituitaries and correspond to two polyadenylation signal sites in red drum SL cDNA at nucleotide positions 1554 and 1270. Levels of the SL I mRNA were 2- to 4-fold higher in pituitaries of blind red drum and intact fish kept under constant darkness for 1 week than those in control fish sampled during the light phase of the light–dark cycle. Similarly, pituitary levels of SL II mRNA were 9-fold higher in blind fish and 1.6- to 4-fold higher in intact fish kept under constant darkness than in the control fish. Furthermore, these changes in mRNA levels in pituitaries were accompanied by more than 10-fold increases in SL protein concentrations in plasma. The finding that the absence of light perception for extended periods leads to dramatic increases in SL mRNA expression as well as SL secretion in red drum provides further evidence that illumination levels and SL physiology are intimately related in this species.