Biochemical and Biophysical Research Communications
Identification of novel genes expressed during spermatogenesis in stage-synchronized rat testes by differential display
Section snippets
Materials and methods
Preparation of synchronized rat testes. Male, 20-day-old, Sprague–Dawley rats were fed with a diet deficient in vitamin A (ICN, Cleveland, OH) (Groups A–D, n=4) or normal rat chow (Group E, n=4) for 60–70 days. All animals were held in Sealsafe IVC individually ventilated cages (Tecniplast, Buguggiate, Italy). The ethical guidelines set forth by the University’s Committee on Using Live Animals for Teaching and Research were adhered to strictly. After histological verification of spermatogenic
Synchronization of VAD testes
After treating 20-day-old SD rats with a VAD diet for 2 months, the animals showed signs of vitamin A deficiency, which included huffed fur, hairlessness, weight loss or cessation of growth, and discharges from the eyes. The representative histology of the testes from VAD rats after designated PVA (retinol) treatment is shown in Fig. 1. At Day 0 PVA, the rat testis contained mainly type A spermatogonia and few preleptotene spermatocytes just before retinol replacement (Fig. 1A). This indicated
Discussion
To elucidate the molecular mechanism regulating the developmental process of male gametes, we employed the mRNA differential display [4] to dissect the molecular mechanisms governing spermatogenesis. This approach is a well-established method that is supplementary to the conventional subtractive hybridization technique and has been used to study the gene expression during spermatogenesis in mice [12], [13], rat [14], [15], [16], and male infertility in human [17]. The present study utilizes the
Acknowledgements
This research was supported by a public-funded grant (HKU7272/01M) to J.M.L. from the Research Grants Council of Hong Kong. We thank Miss Pauline Leung, Mr. Andy Leung, and Eric Leung for technical assistance and Mr. Cheuk Hung Lee for editorial comments.
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Present address: Microbiology and Tumor Biology Centre, MTC Box 280, Karolinska Institute, Stockholm S-171 77, Sweden.