Localization and characterization of an orphan receptor, guanylyl cyclase-G, in mouse testis and sperm

Yen Hua Huang, Chih Chun Wei, Yueh Hsing Su, Bo Tsung Wu, Yi Yun Ciou, Cheng Fen Tu, Trevor G. Cooper, Ching Hei Yeung, Sin Tak Chu, Ming Tzu Tsai, Ruey Bing Yang

研究成果: 雜誌貢獻文章

14 引文 (Scopus)

摘要

We recently identified a novel testis-enriched receptor guanylyl cyclase (GC) in the mouse, designated mGC-G. To further investigate its protein expression and function, we generated a neutralizing antibody specifically against the extracellular domain of this receptor. RT-PCR and immunohistochemical analyses show that mGC-G is predominantly expressed from round spermatids to spermatozoa in mouse testis at both the mRNA and protein levels. Flow cytometry and confocal immunofluorescence reveal that mGC-G is a cell surface protein restricted to the plasma membrane overlying the acrosome and midpiece of the flagellum in mature sperm. Interestingly, Western blot analysis demonstrates that testicular mGC-G is approximately 180 kDa but is subject to limited proteolysis during epididymal sperm transport, resulting in a smaller fragment tethered on the mature sperm surface. On Fluo-3 cytometrical analysis and computer-assisted sperm assay, we found that serum albumin-induced elevation of sperm intracellular Ca 2+ concentration, protein tyrosine phosphorylation, and progressive motility associated with capacitation are markedly reduced by preincubation of the anti-mGC-G neutralizing antibody. Together, these results indicate that mGC-G is proteolytically modified in mature sperm membrane and suggest that mGC-G-mediated signaling may play a critical role in gamete/reproductive biology.

原文英語
頁(從 - 到)4792-4800
頁數9
期刊Endocrinology
147
發行號10
DOIs
出版狀態已發佈 - 2006

指紋

Spermatozoa
Testis
Neutralizing Antibodies
Guanylate Cyclase-Coupled Receptors
Sperm Transport
Acrosome
Proteins
Spermatids
Flagella
Germ Cells
Serum Albumin
Proteolysis
Fluorescent Antibody Technique
Tyrosine
mouse guanylyl cyclase-G
Flow Cytometry
Membrane Proteins
Western Blotting
Phosphorylation
Cell Membrane

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

引用此文

Localization and characterization of an orphan receptor, guanylyl cyclase-G, in mouse testis and sperm. / Huang, Yen Hua; Wei, Chih Chun; Su, Yueh Hsing; Wu, Bo Tsung; Ciou, Yi Yun; Tu, Cheng Fen; Cooper, Trevor G.; Yeung, Ching Hei; Chu, Sin Tak; Tsai, Ming Tzu; Yang, Ruey Bing.

於: Endocrinology, 卷 147, 編號 10, 2006, p. 4792-4800.

研究成果: 雜誌貢獻文章

Huang, YH, Wei, CC, Su, YH, Wu, BT, Ciou, YY, Tu, CF, Cooper, TG, Yeung, CH, Chu, ST, Tsai, MT & Yang, RB 2006, 'Localization and characterization of an orphan receptor, guanylyl cyclase-G, in mouse testis and sperm', Endocrinology, 卷 147, 編號 10, 頁 4792-4800. https://doi.org/10.1210/en.2005-1476
Huang, Yen Hua ; Wei, Chih Chun ; Su, Yueh Hsing ; Wu, Bo Tsung ; Ciou, Yi Yun ; Tu, Cheng Fen ; Cooper, Trevor G. ; Yeung, Ching Hei ; Chu, Sin Tak ; Tsai, Ming Tzu ; Yang, Ruey Bing. / Localization and characterization of an orphan receptor, guanylyl cyclase-G, in mouse testis and sperm. 於: Endocrinology. 2006 ; 卷 147, 編號 10. 頁 4792-4800.
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abstract = "We recently identified a novel testis-enriched receptor guanylyl cyclase (GC) in the mouse, designated mGC-G. To further investigate its protein expression and function, we generated a neutralizing antibody specifically against the extracellular domain of this receptor. RT-PCR and immunohistochemical analyses show that mGC-G is predominantly expressed from round spermatids to spermatozoa in mouse testis at both the mRNA and protein levels. Flow cytometry and confocal immunofluorescence reveal that mGC-G is a cell surface protein restricted to the plasma membrane overlying the acrosome and midpiece of the flagellum in mature sperm. Interestingly, Western blot analysis demonstrates that testicular mGC-G is approximately 180 kDa but is subject to limited proteolysis during epididymal sperm transport, resulting in a smaller fragment tethered on the mature sperm surface. On Fluo-3 cytometrical analysis and computer-assisted sperm assay, we found that serum albumin-induced elevation of sperm intracellular Ca 2+ concentration, protein tyrosine phosphorylation, and progressive motility associated with capacitation are markedly reduced by preincubation of the anti-mGC-G neutralizing antibody. Together, these results indicate that mGC-G is proteolytically modified in mature sperm membrane and suggest that mGC-G-mediated signaling may play a critical role in gamete/reproductive biology.",
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AU - Ciou, Yi Yun

AU - Tu, Cheng Fen

AU - Cooper, Trevor G.

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AU - Chu, Sin Tak

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