Initial Characterization of an In Vitro Rabbit Sperm Activation Assay

Details

• Personal Author:
  Breitenstein, Michael J. ; Brown DB ; Hillman GR ; Maeker BJ ; Sawyer DE ; Schrader, Steven M.
• Description:
  Previous studies have shown that both human and rat sperm nuclei can be activated in vitro using a cytoplasmic extract from Xenopus laevis frog eggs. Activated nuclei undergo chromatin decondensation, DNA synthesis, and recondensation (human only). To complement these systems, we are developing a rabbit sperm activation assay. Lysolecithin-permeabilized rabbit sperm were incubated in 3H-TTP-spiked X. laevis frog egg extract. Decondensation was assessed visually using phase-contrast microscopy and quantitatively using an image analysis system. DNA synthesis was assessed using autoradiography. Ejaculates from three different rabbits were used for these studies. Activated sperm nuclei from two of the rabbits decondensed to about 11 times their original size at a rate of approximately 175 U/min. (arbitrary units). Interestingly, multiple ejaculates from one rabbit repeatedly displayed attenuated decondensation (7.5X size increase and 114 U/min.); the reason for this is being investigated. Greater than 95% of the activated sperm nuclei consistently replicate DNA. We plan to use the rabbit sperm activation assay to perform in vivo toxicology studies on toxic agents known to bind sperm chromatin. This assay will complement the rat sperm activation assay and allow for multispecies toxicology studies evaluating sperm nuclear activation as an endpoint. [Description provided by NIOSH]
• Subjects:
  Animals Environmental Monitoring Hazardous Substances Laboratories Occupational Health Safety Urogenital Diseases Urogenital System
• Keywords:
  Animal-studies Exposure-assessment Exposure-levels Exposure-limits Laboratory-animals Reproductive-effects Reproductive-hazards Reproductive-system Reproductive-system-disorders Toxic-materials Toxins

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• ISSN:
  1096-6080
• Document Type:
  Text
• Genre:
  Abstract or Summary
• Place as Subject:
  Ohio ; OSHA Region 5 ; OSHA Region 6 ; Texas
• CIO:
  National Institute for Occupational Safety and Health (NIOSH)
• Division:
  DBBS - Division of Biomedical and Behavioral Sciences ; EID - Education and Information Division
• Topic:
  Workplace Safety & Health
• Location:
  North America
• Volume:
  36
• Issue:
  1
• NIOSHTIC Number:
  nn:20030614
• Citation:
  Toxicologist 1997 Mar; 36(1)(Pt 2):360
• Federal Fiscal Year:
  1997
• Peer Reviewed:
  False
• Part Number:
  2
• Source Full Name:
  The Toxicologist. Society of Toxicology 36th Annual Meeting, March 9-13, 1997, Cincinnati, Ohio
• Collection(s):
  National Institute for Occupational Safety and Health
• Main Document Checksum:
  urn:sha-512:c1dd173e9d9493403399b4bee4fbcf23fa2c5fafa2005a3129f98d9eb67b84f2905f065a30f135edc5d33e0823ebcfa4d3ebf5a60fe2346265e1b4f181132f2c
• Download URL:
  https://stacks.cdc.gov/view/cdc/191163/cdc_191163_DS1.pdf
• File Type:
  [PDF - 157.27 KB ]