Identification of phospholipase C zeta in normospermic and teratospermic domestic cat sperm

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• 作者：Ana Izabel S. Balbin Villaverde ; Eduardo G. Fioratti ; Rafael A. Fissore ; Changli He ; Hoi Chang Lee ; Fabiana F. Souza ; Fern ; a C. L ; im-Alvarenga ; Maria Denise Lopes
• 关键词：Fertilization ; Oocyte activation ; Immunoblotting ; Two-dimensional PAGE ; PLC¦Æ
• 刊名：Theriogenology
• 出版年：2013
• 出版时间：15 October, 2013
• 年：2013
• 卷：80
• 期：7
• 页码：722-729
• 全文大小：1011 K

文摘

In mammalian species, oocyte activation is initiated by oscillations in the intracellular concentration of free calcium ([Ca²⁺]_i), which are also essential to allow embryonic development. To date, evidence supporting the hypothesis that a sperm factor is responsible for initiating oocyte activation has been presented in various mammalian species. Among the possible candidates to be the active sperm factor is the novel sperm-specific phospholipase C ¦Æ (PLC¦Æ), which besides its testis-specific expression is capable of initiating [Ca²⁺]_i oscillations. In this study, we investigated the presence of PLC¦Æ in the sperm of the domestic cat and whether normospermic and teratospermic cats differ in their PLC¦Æ expression. Immunoblotting with anti-PLC¦Æ antibodies confirmed the presence of an immunoreactive band of ¡«70 kDa in whole sperm lysates of domestic cat as well as in both soluble and ¡°insoluble¡± fractions from this sperm. Additional immunoreactive bands, probably C- and N-terminal truncated versions of PLC¦Æ, were also visualized in the soluble sperm fractions. Interestingly, immunoreactivity of PLC¦Æ was detectable in teratospermic sperm, although with slightly less intensity than in normospermic sperm. In conclusion, domestic cat sperm express PLC¦Æ in both cytosolic and high-pH fractions, which is consistent with data in other mammals. Sperm from teratospermic cats also express PLC¦Æ, albeit at reduced concentrations, which may affect the fertility of these males.