Rajesh Angireddy, Yenugu Suresh
Department of Animal Biology, University of Hyderabad, Hyderabad 500 046, India.
Department of Animal Biology, University of Hyderabad, Hyderabad 500 046, India.
J Reprod Immunol. 2015 Aug;110:117-29. doi: 10.1016/j.jri.2015.02.009. Epub 2015 Apr 10.
Evaluating the immunocontraceptive potential of sperm-bound proteins is an active area of investigation. In this study, we analyzed the role of prostate- and testes-expressed (PATE) and PATE-F proteins in sperm function. Capacitation was measured as a function of tyrosine phosphorylation of sperm membrane proteins. Ionophore-induced acrosome reaction was assessed by measuring the fluorescence intensity of calcium-bound Fluo 3-AM and sperm-bound PNA-FITC in a flow cytometer. Rat spermatozoa subjected to capacitation and acrosome reaction in vitro displayed changes in the PATE and PATE-F protein localization on their surface, indicating the role of these proteins in sperm function. Capacitation and ionophore-induced acrosome reaction in vitro were inhibited in spermatozoa pre-incubated with antiserum raised in rabbit against PATE or PATE-F. Male rats were immunized with PATE proteins to assess their role in sperm function and fecundity. Antibody titer in the serum, testicular, and epididymal fluid was measured by ELISA. The motility parameters were recorded using CASA. High antibody titer was observed in serum, epididymal, and testicular fluid in rats immunized with PATE or PATE-F protein. Immunization did not cause any structural damage and inflammation in the testis and epididymis. PATE and PATE-F antisera obtained from the immunized rats inhibited acrosome reaction. Motility parameters, capacitation, acrosome reaction, and fecundity were compromised in PATE-F-immunized rats, whereas the same were not affected in rats immunized with PATE. These results suggest that PATE-F might play an important role in sperm function and fecundity and can be explored further to determine its immunocontraceptive potential.
评估精子结合蛋白的免疫避孕潜力是一个活跃的研究领域。在本研究中，我们分析了前列腺和睾丸表达（PATE）蛋白及PATE - F蛋白在精子功能中的作用。通过精子膜蛋白酪氨酸磷酸化来衡量获能情况。离子载体诱导的顶体反应通过在流式细胞仪中测量钙结合的Fluo 3 - AM和精子结合的PNA - FITC的荧光强度来评估。体外经历获能和顶体反应的大鼠精子在其表面的PATE和PATE - F蛋白定位上出现变化，表明这些蛋白在精子功能中发挥作用。用兔抗PATE或PATE - F抗血清预孵育的精子，其体外获能和离子载体诱导的顶体反应受到抑制。用PATE蛋白免疫雄性大鼠以评估其在精子功能和生育力中的作用。通过ELISA测量血清、睾丸和附睾液中的抗体滴度。使用CASA记录运动参数。在用PATE或PATE - F蛋白免疫的大鼠的血清、附睾和睾丸液中观察到高抗体滴度。免疫并未在睾丸和附睾中引起任何结构损伤和炎症。从免疫大鼠获得的PATE和PATE - F抗血清抑制顶体反应。PATE - F免疫的大鼠的运动参数、获能、顶体反应和生育力受到损害，而PATE免疫的大鼠则不受影响。这些结果表明，PATE - F可能在精子功能和生育力中发挥重要作用，可进一步探索以确定其免疫避孕潜力。
