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Expression Analysis of Glutathione Peroxidase Genes in the Stage-Specific Seminiferous Tubules of Mice Excised by a Laser Capture Microdissection KCI 등재
Laser Capture Microdissection으로 절제된 마우스의 특정 단계별 정세관에서 Glutathione Peroxidase 유전자의 발현 분석
pp.99-105
정세관은 매우 복잡한 조직으로 마우스의 정자 발생 과정은 12단계로 구성되어 있다. Glutathione peroxidase(GPx)는 glutathione을 이용하여 과산화물(hydroperoxide)을 환원시키는 대표적인 항산화효소로서 포유류 정자 발생 과정에 관여하는 것으로 알려져 있다. 본 연구에서는 laser capture microdissection(LCM)을 이용하여 마우스 정소에서 발생 단계별로 정세관을 채취하여 real-time PCR로
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.
