Identification and characterization of an oocyte factor required for sperm decondensation in pig

Reproduction. 2014 Oct;148(4):367-75. doi: 10.1530/REP-14-0264. Epub 2014 Jul 16.

Abstract

Mammalian oocytes possess factors to support fertilization and embryonic development, but knowledge on these oocyte-specific factors is limited. In the current study, we demonstrated that porcine oocytes with the first polar body collected at 33 h of in vitro maturation sustain IVF with higher sperm decondensation and pronuclear formation rates and support in vitro development with higher cleavage and blastocyst rates, compared with those collected at 42 h (P<0.05). Proteomic analysis performed to clarify the mechanisms underlying the differences in developmental competence between oocytes collected at 33 and 42 h led to the identification of 18 differentially expressed proteins, among which protein disulfide isomerase associated 3 (PDIA3) was selected for further study. Inhibition of maternal PDIA3 via antibody injection disrupted sperm decondensation; conversely, overexpression of PDIA3 in oocytes improved sperm decondensation. In addition, sperm decondensation failure in PDIA3 antibody-injected oocytes was rescued by dithiothreitol, a commonly used disulfide bond reducer. Our results collectively report that maternal PDIA3 plays a crucial role in sperm decondensation by reducing protamine disulfide bonds in porcine oocytes, supporting its utility as a potential tool for oocyte selection in assisted reproduction techniques.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cells, Cultured
  • Female
  • Fertilization in Vitro
  • Gene Expression Regulation, Developmental
  • Gene Expression Regulation, Enzymologic
  • In Vitro Oocyte Maturation Techniques
  • Male
  • Oocytes / enzymology*
  • Paracrine Communication*
  • Protein Disulfide-Isomerases / genetics
  • Protein Disulfide-Isomerases / metabolism*
  • Proteomics / methods
  • RNA, Messenger / metabolism
  • Signal Transduction
  • Sperm Injections, Intracytoplasmic
  • Sperm-Ovum Interactions*
  • Spermatozoa / enzymology*
  • Swine
  • Time Factors
  • Transfection

Substances

  • RNA, Messenger
  • Pdia3 protein, mouse
  • Protein Disulfide-Isomerases