cDNA sequence and spatio-temporal expression of prolactin in the orange-spotted grouper, Epinephelus coioides

Abstract

The cDNA sequence encoding the prolactin (PRL) gene of the orange-spotted grouper was obtained by randomly sequencing its pituitary cDNA library. The full-length cDNA of orange-spotted grouper PRL consists of 1342 bp. The open reading frame encodes a protein of 212 amino acids, which consists of a putative signal peptide of 24 residues and a mature protein of 188 amino acids. Northern blot analysis identified a single transcript of 1.4 kb in the pituitary gland, indicating perhaps the presence of only one form of PRL in the orange-spotted grouper. The expression of PRL gene in different tissues and during embryonic development of orange-spotted grouper was determined using one-step RT-PCR followed by Southern blot analysis. In addition to the pituitary gland, high levels of PRL gene expression were detected in the olfactory bulb, forebrain, midbrain, hypothalamus, hindbrain, medulla, gill, spleen, and adipose tissue. Very low levels of expression were detected in the kidney, liver, and blood cells. The PRL gene was also expressed in the gonads, with the higher level of expression observed in the stage 3 as compared to stage 2 ovary. During embryonic development, mRNA for PRL was detected at all time points examined except at 2h 15min post-fertilization, and its expression peaked at 9h post-fertilization, at the time of neurulation. Results from the current study suggest that PRL may have important paracrine or autocrine roles in diverse tissue types, including the brain and the gonads, and during embryonic development of the orange-spotted grouper.