Laser Capture Microdissection으로 절제된 마우스의 특정 단계별 정세관에서 Glutathione Peroxidase 유전자의 발현 분석

The seminiferous epithelium, with its division into 12 spermatogenic stages in the mouse, is a very complex tissue. Glutathione peroxidase (GPx) is a representative antioxidant enzyme that is capable of reducing organic hydroperoxides to their corresponding hydroxyl compounds utilizing glutathione and is related to the mammalian spermatogenesis. In this study, a real-time PCR was performed in the stage-specific seminiferous tubules of mouse testes excised by a laser capture microdissection (LCM) in order to quantitate the expression levels of a series of GPx genes including cytosolic GPx (cGPx), gastrointestinal GPx (GI-GPx), plasma GPx (pGPx), and phospholipid hydroperoxide GPx (PHGPx). Frozen sections (10 ㎛) were obtained from normal adult mouse testes. LCM was used to capture all the cells that were grouped into stages Ⅰ-Ⅴ, Ⅶ-Ⅷ, and Ⅸ-ⅩI in cross-sections of seminiferous tubules. The expression level of PHGPx mRNA was remarkably higher than those of other GPx mRNAs in mouse testes. During spermatogenesis, the expressions of GI-GPx, pGPx, and PHGPx mRNAs were highest on stages Ⅶ-Ⅷ, began to decrease after stage ⅩI, and showed a lowest level on stage Ⅰ-Ⅴ. However, the expressions of cGPx mRNA were highest on stages Ⅶ-Ⅷ, and showed a lowest level on stage ⅩI-ⅩI. These findings indicate that GPx genes are expressed differentially on mouse spermatogenesis and also LCM can be an useful tool in cellular quantitative analysis of testes.