Abstract
We performed annealing control primer (ACP)-based differential-display reverse transcription-polymerase chain reaction (DDRT-PCR) to isolate differentially expressed genes (DEGs) from the stage IV ovary and ovotestis of the rice field eel, Monopterus albus. Using 20 arbitrary ACP primers, 14 DEG expressed-sequence tags were identified and sequenced. The transcriptional expression of one DEG, G2, was significantly greater in the ovotestis than the stage IV ovary. To understand the role of G2 in sex inversion, G2 cDNA was cloned and semi-RT-PCR, real time PCR were performed during gonad development. The full-length G2 cDNA was 650 base pairs (bp) and it comprised a 5′-untranslated region (UTR) of 82 bp, a 3′-UTR of 121 bp and an open reading frame of 444 bp that encoded a 148-amino acid protein. The expression of G2 was weak during early ovarian development until the stage IV ovary, but expression increased significantly with gonad development. We speculate that G2 may play an important function during sex inversion and testis development in the rice field eel, but the full details of the function of this gene requires further research.
Highlights
In vertebrates, sex can be determined by one of many mechanisms, including chromosomal, polygenic and environmental sex determination
Semi‐quantitative RT‐PCR and real time PCR We identified a differentially expressed gene (DEG) called G2 that had greater levels of transcription in the ovotestis than the stage IV ovary, and this gene was selected to validate the annealing control primer (ACP) DDRT-PCR prescreening approach
Because the paraffin sections showed there were no male tissues in the stage IV ovary tissues while the spermatocytes first appeared in ovotestis, we decided to isolate DEGs from the gonad tissues at these stages
Summary
Sex can be determined by one of many mechanisms, including chromosomal, polygenic and environmental sex determination. In some fish, more than one mechanism can contribute to sex determination and different species exhibit various mechanisms for controlling sexual determination and patterns of sexual differentiation. It is a protogynous freshwater fish that exhibits an adult sex inversion from functional females to males (Liu 1944; Liem 1963; Chan and Phillips 1967; Xiao and Liu 1995b). Spermatogonia begin to multiple on the germinal fold and form spermatocysts This is the intersex development stage, and the male individual develops from the female. The male development of M. albus can be divided into different stages. This eel is commonly studied because of these unique physical characteristics
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