Abstract
BackgroundInhibins are dimeric gonadal protein hormones that negatively regulate pituitary FSH synthesis and secretion. Inhibin B is produced by testicular Sertoli cells and is the primary circulating form of inhibin in most adult male mammals. Inhibin B is comprised of the inhibin alpha subunit disulfide-linked to the inhibin/activin betaB subunit. Here we describe the cloning of the cDNAs encoding these subunits from adult rhesus monkey testis RNA.MethodsThe subunit cDNAs were cloned by a combination of reverse transcriptase polymerase chain reaction (RT-PCR) and 5' rapid amplification of cDNA ends (RACE) RT-PCR from adult rhesus monkey testis RNA.ResultsBoth the inhibin alpha and betaB subunit nucleotide and predicted protein sequences are highly conserved with other mammalian species, particularly with humans. During the course of these investigations, a novel inhibin alpha mRNA isoform was also identified. This form, referred to as rhesus monkey inhibin alpha-variant 2, appears to derive from both alternative transcription initiation as well as alternative splicing. rmInhibin alpha-variant 2 is comprised of a novel 5' exon (exon 0), which is spliced in-frame with exon 2 of the conventional inhibin alpha isoforms (variant 1). Exon 1 is skipped in its entirety such that the pro-alpha and part of the alpha N regions are not included in the predicted protein. rmInhibin alpha -variant 2 is of relatively low abundance and its biological function has not yet been ascertained.ConclusionThe data show that the predicted inhibin B protein is very similar between monkeys and humans. Therefore, studies in monkeys using recombinant human inhibins are likely to reflect actions of the homologous ligands. In addition, we have observed the first inhibin alpha subunit mRNA variant. It is possible that variants will be observed in other species as well and this may lead to novel insights into inhibin action.
Highlights
Inhibins are dimeric gonadal protein hormones that negatively regulate pituitary FSH synthesis and secretion
The inhibin α cDNA was cloned from rhesus monkey testis using a combination of reverse transcriptase polymerase chain reaction (RT-PCR) based approaches
We describe the cloning of the inhibin B subunit cDNAs from testis of the adult rhesus monkey
Summary
Inhibins are dimeric gonadal protein hormones that negatively regulate pituitary FSH synthesis and secretion. Reproductive Biology and Endocrinology 2004, 2:71 http://www.rbej.com/content/2/1/71 do so in discordant fashion across the reproductive cycle [3,4,5,6,7,8,9,10,11,12,13,14,15] One exception to this general pattern is in rams, where inhibin A appears to be the primary circulating form [16]. Because inhibin B is the biologically relevant ligand in male primates, this has placed some constraints on our understanding of inhibin action in these animals For this reason, we cloned the inhibin B subunit cDNAs from adult monkey testis as a requisite first step to producing recombinant monkey inhibin B. We describe the new transcript called rhesus monkey inhibin α-variant 2
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