Flow Cytometry of Fine Needle Aspirations of the Sprague-Dawley Rat Testis: Defining Normal Maturation and the Effects of Multiple Biopsies

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Abstract
Quantitative assessment of spermatogenesis by flow cytometry reproducibly correlates with histological examination. Flow cytometry of fine needle aspirates from the testis was used to analyze normal spermatogenesis of Sprague-Dawley rats. Aged matched male weanlings were divided into 3 groups: group 1-5 rats underwent bilateral percutaneous aspiration of the testes with a 22 gauge Chiba needle on postpartum days 19, 24, 30 and 40; group 2-5 rats aspirated on days 24, 30 and 40, and group 3-4 rats aspirated on days 30 and 40. This sequential approach allowed for evaluation of normal spermatogenesis and the effects of repeated fine needle aspiration on spermatogenesis. Deoxyribonucleic acid distribution analysis by flow cytometry correlates with haploid (1N), diploid (2N), tetraploid (4N) and S phase cell populations. These cell populations were evaluated and comparisons among each group were made at all biopsy times. No significant differences in mean 2N, 4N or S phase cell populations in single versus repeat biopsied testes were detected. However, there was a significant increase in the 1N population at 30 days post partum in the repeat biopsy group (p = 0.037), which normalized by day 40. This increase in 1N corresponds to the beginning of meiosis, which is maximal between postpartum days 30 and 40, and occurs earlier in the repeat biopsied testis. Repeat fine needle aspiration of the testis can be performed without significantly affecting spermatogenesis in the "weanling" rat. This provides a useful technique in the future investigations of the time related effects of varicocele, chemotherapy and toxicologic drugs on spermatogenesis.

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