کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10934021 | 1093877 | 2005 | 17 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Functional, biochemical, and chromatographic characterization of the complete [Ca2+]i oscillation-inducing activity of porcine sperm
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موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
بیولوژی سلول
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چکیده انگلیسی
A cytosolic sperm protein(s), referred to as sperm factor (SF), is delivered into eggs by the sperm during mammalian fertilization to induce repetitive increases in the intracellular concentration of free Ca2+ ([Ca2+]i) that are referred to as [Ca2+]i oscillations. [Ca2+]i oscillations are essential for egg activation and early embryonic development. Recent evidence shows that the novel sperm-specific phospholipase C (PLC), PLCζ, may be the long sought after [Ca2+]i oscillation-inducing SF. Here, we demonstrate the complete extraction of SF from porcine sperm and show that regardless of the method of extraction a single molecule/complex appears to be responsible for the [Ca2+]i oscillation-inducing activity of these extracts. Consistent with this notion, all sperm fractions that induced [Ca2+]i oscillations, including FPLC-purified fractions, exhibited high in vitro PLC activity at basal Ca2+ levels (0.1-5 μM), a hallmark of PLCζ. Notably, we detected immunoreactive 72-kDa PLCζ in an inactive fraction, and several fractions capable of inducing oscillations were devoid of 72-kDa PLCζ. Nonetheless, in the latter fractions, proteolytic fragments, presumably corresponding to cleaved forms of PLCζ, were detected by immunoblotting. Therefore, our findings corroborate the hypothesis that a sperm-specific PLC is the main component of the [Ca2+]i oscillation-inducing activity of sperm but provide evidence that the presence of 72-kDa PLCζ does not precisely correspond with the Ca2+ releasing activity of porcine sperm fractions.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Developmental Biology - Volume 285, Issue 2, 15 September 2005, Pages 376-392
Journal: Developmental Biology - Volume 285, Issue 2, 15 September 2005, Pages 376-392
نویسندگان
Manabu Kurokawa, Ken-ichi Sato, Hua Wu, Changli He, Christopher Malcuit, Samuel J. Black, Kiyoko Fukami, Rafael A. Fissore,