MOLECULAR REPRODUCTION AND DEVELOPMENT 69:411–418 (2004) Immunolocalization of Bovine Sperm Protease BSp120 by Light and Electron Microscopy During Capacitation and the Acrosome Reaction: Its Role in In Vitro Fertilization

نویسندگان

  • ANDREINA CESARI
  • JORGE J. SÁNCHEZ
  • JUAN C. BIANCOTTI
  • MÓNICA H. VAZQUEZ-LEVIN
  • GERMÁN KAISER
  • GUSTAVO A. PALMA
  • RICARDO ALBERIO
  • AMANDA E. VINCENTI
  • MIGUEL W. FORNÉS
چکیده

Mammalian fertilization involves various steps in which the participation of specific enzymes has been demonstrated by numerous studies. Acrosin is one of the most widely acrosomal protease in mammalian spermatozoa studied, including bovine; however, other proteases have also been described. A new trypsin-like serine protease named bovine serine protease of 120 kDa (BSp120) and its pre-cursor BSp66 (66 kDa) were identified in bovine spermatozoa. Cytological and ultrastructural immunolocalization studies on BSp120 were performed in live and fixed cells. Immunoflorescence assays with specific polyclonal antibodies revealed localization of BSp120 on the sperm head, with a signal homogeneously distributed over the acrosome resembling a horseshoe. After the acrosome reaction, sperm showed a patchy pattern in the acrosomal cap. Immune electron microscopy analysis indicated that BSp120 is located over the head plasma membrane of capacitated spermatozoa and acrosome reacting spermatozoa. To assess BSp120 function in sperm-oocyte interaction, in vitro fertilization studies were conducted. Oocytes were incubated with spermatozoa pre-treated with antiBSp120, anti-guinea pig acrosin, and anti-BSp120 plus anti-guinea pig acrosin. Pre-treatment of bovine spermatozoa with antibodies towards each protein did not significantly modify fertilization rates. However, when both anti-acrosin and anti-BSp120 antibodies were simultaneously added, there was a significant decrease in the fertilization rate, suggesting that both enzymes may be required for fertilization. Altogether, the results from the present study described the localization of BSp120 over the acrosome of bovine sperm, and suggest its involvement in fertilization. Mol. Reprod. Dev. 69: 411–418, 2004. 2004 Wiley-Liss, Inc.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

O-8: Molecular Mechanisms of Membrane Fusion Involved in Fertilization

Background: Assisted fertilization procedures are a currently widespread practice to regulate reproduction in humans and animals. The arising question is why the human being manipulating gametes to generate new individuals, if we do not understand yet the molecular mechanism of fertilization?. Successful completion of fertilization in mammals is dependent on three membrane fusion events: 1. the...

متن کامل

Roles of Intracellular Cyclic AMP Signal Transduction in the Capacitation and Subsequent Hyperactivation of Mouse and Boar Spermatozoa

It is not until accomplishment of a variety of molecular changes during the transit through the female reproductive tract that mammalian spermatozoa are capable of exhibiting highly activated motility with asymmetric whiplash beating of the flagella (hyperactivation) and undergoing acrosomal exocytosis in the head (acrosome reaction). These molecular changes of the spermatozoa are collectively ...

متن کامل

Physiology and endocrinology symposium: evidence that oviduct secretions influence sperm function: a retrospective view for livestock.

The mammalian oviduct has long been recognized as an organ essential for successful reproduction. Bovine, ovine, porcine, and equine animal models have offered clear advantages for oviduct study related to gamete physiology, fertilization, and early embryonic development. Livestock species are amenable to surgical alteration of the reproductive tract, estrous cycle manipulation, gamete cryopres...

متن کامل

Physiology and Endocrinology: Sperm-Oviduct Interactions in Livestock and Poultry

The mammalian oviduct has long been recognized as an organ essential to the success of reproduction. Bovine, ovine, porcine and equine animal models have offered clear advantages for oviduct study related to gamete physiology, fertilization and early embryo development. Livestock species are amenable to surgical alteration of the reproductive tract, estrous cycle manipulation, gamete cryopreser...

متن کامل

Premature capacitation of frozen-thawed spermatozoa from subfertile Japanese black cattle.

Artificial insemination (AI) subfertility is an indication of failure of AI with frozen-thawed sperm classified as normal by conventional semen examination. Recently, 8 AI-subfertile Japanese Black cattle (S1-S8) were identified using the routine AI test or in vivo fertilization test, which included AI with frozen-thawed sperm of superovulated females and subsequent non-surgical recovery of pre...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:

دوره   شماره 

صفحات  -

تاریخ انتشار 2004