Herrero María Belén, Mandal Arabinda, Digilio Laura C, Coonrod Scott A, Maier Bernhard, Herr John C
Center for Research in Contraceptive and Reproductive Health, Department of Cell Biology, P.O. Box 800732, University of Virginia, Charlottesville, VA 22908, USA.
Dev Biol. 2005 Aug 1;284(1):126-42. doi: 10.1016/j.ydbio.2005.05.008.
This study demonstrates the retention of mouse sperm lysozyme-like protein (mSLLP1) in the equatorial segment of spermatozoa following the acrosome reaction and a role for mSLLP1 in sperm-egg binding and fertilization. Treatment of cumulus intact oocytes with either recmSLLP1 or its antiserum resulted in a significant (P < or = 0.05) inhibition of fertilization. Co-incubation of zona-free mouse oocytes with capacitated mouse spermatozoa in the presence of varying concentrations of anti-recmSLLP1 serum or recmSLLP1 also inhibited sperm-oolemma binding. A complete inhibition of binding and fusion of spermatozoa to the oocyte occurred at 12.5 muM concentration of recmSLLP1, while conventional chicken and human lysozymes did not block sperm-egg binding. mSLLP1 showed receptor sites in the perivitelline space as well as on the microvillar region of the egg plasma membrane. The retention of mSLLP1 in the equatorial segment of acrosome-reacted sperm, the inhibitory effects of both recmSLLP1 and antibodies to SLLP1 on in vitro fertilization with both cumulus intact and zona-free eggs, and the definition of complementary SLLP1-binding sites on the egg plasma membrane together support the hypothesis that a c lysozyme-like protein is involved in the binding of spermatozoa to the egg plasma membrane during fertilization.
本研究证明,顶体反应后小鼠精子溶菌酶样蛋白(mSLLP1)保留在精子的赤道段,且mSLLP1在精卵结合和受精过程中发挥作用。用重组mSLLP1或其抗血清处理完整卵丘的卵母细胞,会导致受精受到显著抑制(P≤0.05)。在不同浓度的抗重组mSLLP1血清或重组mSLLP1存在的情况下,将去透明带的小鼠卵母细胞与获能的小鼠精子共同孵育,也会抑制精子与卵质膜的结合。当重组mSLLP1浓度达到12.5μM时,精子与卵母细胞的结合和融合被完全抑制,而传统的鸡和人溶菌酶不会阻断精卵结合。mSLLP1在卵周隙以及卵质膜微绒毛区域显示出受体位点。mSLLP1在顶体反应精子的赤道段的保留、重组mSLLP1和SLLP1抗体对完整卵丘和去透明带卵体外受精的抑制作用,以及卵质膜上互补的SLLP1结合位点的确定,共同支持了一种假说,即一种溶菌酶样蛋白在受精过程中参与精子与卵质膜的结合。