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KMID : 0894520100140020099
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Development & Reproduction
2010 Volume.14 No. 2 p.99 ~ p.105
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Expression Analysis of Glutathione Peroxidase Genes in the Stage-Specific Seminiferous Tubules of Mice Excised by a Laser Capture Microdissection
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Yon Jung-Min
Lin Chun-Mei
Park Jung-Hoon
Hong Min-Ki
Jung A-Young
Kim Mi-Ra
Baek In-Jeoung
Lee Beom-Jun
Nam Sang-Yoon
Yun Young-Won
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Abstract
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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.
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KEYWORD
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Glutathione peroxidase, Spermatogenesis, Laser capture microdissection, Real-time PCR
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