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Molecular cloning of prepro-thyrotropin-releasing hormone cDNA from medaka (Oryzias latipes)
Yasuhiro Aoki1, Tomohiro Masuda, Masayuki Iigo
1Department of Biotechnology, United Graduate School of Agricultural Science, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-cho, Fuchu-shi, Tokyo 183-8509, Japan.
Abstract:
The cDNA encoding prepro-thyrotropin-releasing hormone (ppTRH) in a teleost, medaka (Oryzias latipes) was isolated and characterized. The medaka ppTRH cDNA codes for 270 amino acid residues including eight TRH progenitor sequences (-Lys/Arg-Arg-Gln-His-Pro-Gly-Lys/Arg-Arg-). In silico analyses of the medaka genome database predicted that the structure of the medaka ppTRH gene is similar to the ppTRH genes of the other vertebrate species studied to date; consisting of three exons and two introns. Identity of the medaka ppTRH with the other vertebrates is rather low except the sockeye salmon. A molecular phylogenic tree showed that the ppTRH sequences reflected the predicted pattern of species classification. RT-PCR analysis demonstrated ppTRH gene expression in the brain and retina. These results gave some insight into the molecular evolution of ppTRH and physiological functions of TRH in vertebrates.
Insights
Researchers isolated and characterized the medaka prepro-thyrotropin-releasing hormone (ppTRH) cDNA. This study provides insights into the molecular evolution of ppTRH and the physiological functions of TRH in vertebrates.
Area of Science:
- Molecular Biology
- Evolutionary Biology
- Genomics
Background:
- Thyrotropin-releasing hormone (TRH) plays crucial roles in vertebrate physiology.
- Understanding the evolutionary conservation and divergence of TRH across species is important.
Purpose of the Study:
- To isolate and characterize the cDNA encoding prepro-thyrotropin-releasing hormone (ppTRH) in the teleost medaka (Oryzias latipes).
- To investigate the gene structure, evolutionary relationships, and expression patterns of medaka ppTRH.
Main Methods:
- cDNA isolation and characterization.
- In silico genome database analysis.
- Molecular phylogenetic tree construction.
- Reverse transcription polymerase chain reaction (RT-PCR) for gene expression analysis.
Main Results:
- Medaka ppTRH cDNA encodes 270 amino acids with eight TRH progenitor sequences.
- The medaka ppTRH gene structure (three exons, two introns) is conserved among vertebrates.
- Medaka ppTRH shows low sequence identity with other vertebrates, except sockeye salmon.
- ppTRH gene expression was detected in the medaka brain and retina.
Conclusions:
- The study elucidates the molecular evolution of ppTRH in medaka.
- Findings contribute to understanding the physiological functions of TRH in vertebrates.
- Comparative analysis highlights evolutionary patterns of ppTRH sequences.
