Proteasomal Interference Prevents Zona Pellucida Penetration and Fertilization in Mammals

Details

• Personal Author:
  Caamano JN ; Day BN ; Manandhar G ; McCauley TC ; Sutovsky M ; Sutovsky P ; Thompson WE
• Description:
  The ubiquitin-proteasome pathway has been implicated in the penetration of ascidian vitelline envelope by the fertilizing spermatozoon (Sawada et al., Proc Natl Acad Sci U S A 2002; 99:1223-1228). The present study provides experimental evidence demonstrating proteasome involvement in the penetration of mammalian zona pellucida (ZP). Using porcine in vitro fertilization as a model, penetration of ZP was completely inhibited by specific proteasomal inhibitors MG-132 and lactacystin. Three commercial rabbit sera recognizing 20S proteasomal core subunits beta-1i, beta-2i, alpha-6, and beta-5 completely blocked fertilization at a very low concentration (i.e., diluted 1/2000 to 1/8000 in fertilization medium). Neither proteasome inhibitors nor antibodies had any effects on sperm-ZP binding and acrosome exocytosis in zona-enclosed oocytes or on fertilization rates in zona-free oocytes, which were highly polyspermic. Consistent with a possible role of ubiquitin-proteasome pathway in ZP penetration, ubiquitin and various alpha and beta type proteasomal subunits were detected in boar sperm acrosome by specific antibodies, immunoprecipitated and microsequenced by MALDI-TOF from boar sperm extracts. Antiubiquitin-immunoreactive substrates were detected on the outer face of ZP by epifluorescence microscopy. This study therefore provides strong evidence implicating the ubiquitin-proteasome pathway in mammalian fertilization and zona penetration. This finding opens a new line of acrosome/ZP research because further studies of the sperm acrosomal proteasome can provide new tools for the management of polyspermia during in vitro fertilization and identify new targets for contraceptive development. [Description provided by NIOSH]
• Subjects:
  Immune System Mass Spectrometry Microbiology Occupational Health Safety Spermatogenesis Urogenital Diseases Urogenital System
• Keywords:
  Fertility Immune-reaction Immune-system In-vitro-study Mammalian-cells Mass-spectrometry Microscopic-analysis Reproductive-system Reproductive-system-disorders Spermatozoa
• ISSN:
  0006-3363
• Document Type:
  Text
• Funding:
  Grant
• Genre:
  Journal Article
• Place as Subject:
  Missouri ; Oregon ; OSHA Region 10 ; OSHA Region 7
• CIO:
  National Institute for Occupational Safety and Health (NIOSH)
• Topic:
  Workplace Safety & Health
• Location:
  North America
• Volume:
  71
• Issue:
  5
• NIOSHTIC Number:
  nn:20029411
• Citation:
  Biol Reprod 2004 Nov; 71(5):1625-1637
• Contact Point Address:
  Department of Animal Science, University of Missouri-Columbia, S141 ASRC, 920 East Campus Dr., Columbia, MO 65211-5300
• Email:
  sutovskyp@missouri.edu
• Federal Fiscal Year:
  2005
• Performing Organization:
  University of Missouri, Columbia, Missouri
• Peer Reviewed:
  True
• Start Date:
  20000930
• Source Full Name:
  Biology of Reproduction
• End Date:
  20020929
• Collection(s):
  National Institute for Occupational Safety and Health
• Main Document Checksum:
  urn:sha-512:e5933ce41ad701848e8d4d5276dfc69bc3a004c435223377c29fa3ffeae1de2ca555924e0bd5832bf434af5211fd0f44a18efb2c7e005f367089149abd7444c5
• Download URL:
  https://stacks.cdc.gov/view/cdc/190396/cdc_190396_DS1.pdf
• File Type:
  [PDF - 1.29 MB ]