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Molecular cloning and regulation of mRNA expression of the thyrotropin β and glycoprotein hormone α subunits in red drum, Sciaenops ocellatus

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Abstract

Full-length cDNAs for thyrotropin β (TSHβ) and glycoprotein hormone α (GSUα) subunits were cloned and sequenced from the red drum (Sciaenops ocellatus). The cDNAs for TSHβ (877 bp) and GSUα (661 bp) yielded predicted coding regions of 126 and 94 amino acid proteins, respectively. Both sequences contain all invariant cysteine and putative glycosylated asparagines characteristic of each as deduced by comparison with other GSUα and TSHβ sequences from representative vertebrate species. Multiple protein sequence alignments show that each subunit shares highest identity (79% for the TSHβ and 86% for the GSUα) with perciform fish. Furthermore, in a single joint phylogenetic analysis, each subunit segregates most closely with corresponding GSUα and TSHβ subunit sequences from closely related fish. Tissue-specific expression assays using RT-PCR showed expression of the TSHβ subunit limited to the pituitary. GSUα mRNA was predominantly expressed in the pituitary but was also detected in the testis and ovary of adult animals. Northern hybridization revealed the presence of a single transcript for both TSHβ and GSUα, each close in size to mRNA transcripts from other species. Dot blot assays from total RNA isolated from S. ocellatus pituitaries showed that in vivo T3 administration significantly diminished mRNA expression of both the TSHβ and GSUα subunits and that goitrogen treatment caused a significant induction of TSHβ mRNA only. Both TSHβ and GSUα mRNA expression in the pituitary varied significantly in vivo over a 24-h period. Maximal expression for both TSHβ and GSUα occurred during the early scotophase in relation to a peak in T4 blood levels previously documented. These results suggest the production of TSH in this species which may serve to drive daily cycles of thyroid activity. Readily quantifiable, variable, and thyroid hormone-responsive pituitary TSH expression, coupled with previously described dynamic daily cycles of circulating T4 and extensive background on the growth, nutrition, and laboratory culture of red drum, suggests that this species will serve as a useful model for experimental studies of the physiological regulation of TSH production.

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Acknowledgments

We thank Dr. Scott Jaques of the Texas Veterinary Medical Diagnostic Laboratory for providing facilities and reagents for performing thyroid hormone radioimmunoassays, Brian Ray and Dr. Delbert Gatlin of Texas A&M University and Gary Jones of the Dow Chemical Company for assistance in obtaining or maintaining fish, and Drs. Suraj Unniappan and Luis Fabian Canosa of the University of Alberta for assistance in developing the molecular biological techniques. The authors also wish to acknowledge the assistance of Dr. Arjun Natesan of Texas A&M during the early stages of this project. Support for this study came from The Texas Advanced Technology Program (project 010366-445), the Texas A&M-CONACYT Collaborative Research Grant Program, College of Science Research Enhancement funds, Texas A&M University, and a mini-grant from McKendree University. Additional support was provided by grants from the Louisiana Board of Regents and NIEHS 5S11 ES9996.

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  1. Kevin A. Leiner

    Present address: Syngenta Biotechnology, Inc., Research Triangle Park, Research Triangle, NC, 27709, USA

Authors and Affiliations

  1. Department of Biology, Texas A&M University, 3258 TAMU, College Station, TX, 77843, USA

    William B. Cohn, Richard A. Jones & Duncan S. MacKenzie

  2. Department of Biological Sciences, Southeastern Louisiana University, 808 North Pine Street, SLU 10736, Hammond, LA, 70402, USA

    Roldan A. Valverde

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  1. William B. Cohn
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  2. Richard A. Jones
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  3. Roldan A. Valverde
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  4. Kevin A. Leiner
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  5. Duncan S. MacKenzie
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Correspondence to William B. Cohn.

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Cohn, W.B., Jones, R.A., Valverde, R.A. et al. Molecular cloning and regulation of mRNA expression of the thyrotropin β and glycoprotein hormone α subunits in red drum, Sciaenops ocellatus . Fish Physiol Biochem 36, 1277–1290 (2010). https://doi.org/10.1007/s10695-010-9408-5

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  • DOI: https://doi.org/10.1007/s10695-010-9408-5

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