Immunization of Mice with Heat-Solubilized Hamster Zonae: Production of Anti-Zona Antibody and Inhibition of Fertility

Section: Introductionmentioning

confidence: 99%

Long-term effects of porcine zonae pellucidae immunocontraception on ovarian function in feral horses (Equus caballus)

Kirkpatrick¹,

Liu

Turner

et al. 1992

“…There was a considerable degree of discrepancy between the two titres of zona precipitate and immunofluorescence in the present study. As suggested by Gwatkin et al (1977) …”

Section: Methodsmentioning

confidence: 83%

“…The specificity of the zona-antigen(s) was further established by using, as the immunogen, cumulus-free mouse eggs (Tsunoda, 1977), isolated mouse zonae pellucidae (Gwatkin, Williams & Carlo, 1977;Tsunoda & Sugie, 1977) or isolated pig zonae pellucidae (Sacco & Palm, 1977). For the purpose of immunological regulation of human fertility, specificity of the zona-antigen(s) must be established with certainty.…”

Section: Introductionmentioning

confidence: 99%

Heteroimmunization with isolated human ova

Takai¹,

Mori²,

Noda³

et al. 1981

“…It is well known that disturbing ZP function by active immunization with heterologous ZP antigens alters fertility in rabbits (Gwatkin & Williams, 1978;Gwatkin et ai, 1980;Wood et al, 1981), dogs (Mahi-Brown et ai, 1982), primates (Gulyas et ai, 1983;Sacco et ai, 1989), horses (Liu et al, 1989;Kirkpatrick et ai, 1990) and mice (Gwatkin et al, 1977). Clinical application of this knowledge to provide methods of immunocontraception for domestic and feral equine populations will ultimately rely upon recombinant techniques (Tyndale-Biscoe, 1991).…”

Section: Discussionmentioning

confidence: 99%

Characterization of equine zona pellucida glycoproteins by polyacrylamide gel electrophoresis and immunological techniques

Miller

Fayrer-Hosken²,

Timmons³

et al. 1992

Summary. This study was designed to explore the composition of the equine zona pellucida (EZP) by one-and two-dimensional polyacrylamide gel electrophoresis (1D-and 2D-PAGE), silver staining and immunoblotting techniques. Antral follicles palpable on frozen\p=n-\thawed equine ovaries were aspirated with a needle and syringe, and the resultant follicular fluid, cellular material and oocytes were pooled. Oocytes were placed in Petri dishes, moved by narrow-bore pipette to droplets of phosphate\x=req-\ buffered saline (PBS) and mechanically cleaned of cumulus cells. The EZP from these collected oocytes was solubilized, and then analysed by 1D-and 2D-PAGE. Silver stained 2D-PAGE of the EZP revealed the presence of three EZP glycoprotein families of apparent molecular mass ranges of 93\p=n-\120 kDa, 73\p=n-\90kDa and 45\p=n-\80kDa.Immunoblot analysis of EZP glycoproteins resolved by 2D-PAGE using rabbit antisera against pig zonae pellucidae (R\g=a\HSPZ) confirmed the presence of three EZP glycoprotein families and established the existence of common epitopes between equine and porcine ZP glycoproteins. Further immunodetection using 2D-PAGE-separated glycoproteins illustrated that the 45\p=n-\80kDa family is recognized by the monoclonal antibody R5, developed against the porcine ZP glycoprotein of molecular mass 55\p=n-\ 120 kDa. Guinea-pig antiserum against endo-\g=b\-galactosidase-treated rabbit ZP 55 kDa glycoprotein (R55K), which specifically recognizes the rabbit ZP glycoprotein with the lowest molecular mass, also recognized the EZP 45\p=n-\80kDa glycoprotein family. Guinea-pig polyclonal antisera developed against total heat-solubilized rabbit ZP (GP\g=a\HSRZ)recognized the 73\p=n-\90kDa EZP glycoprotein family exclusively. After heat solubilization and treatment of EZP with endo-\g=b\-galactosidase to remove polylactosaminoglycans, silver stained 1D-PAGE again demonstrated the presence of three glycoproteins with apparent molecular masses of 60, 75 and 90 kDa. The partially deglycosylated 60 kDa equine glycoprotein is recognized on immunoblot by the monoclonal antibody R5; the 75 kDa EZP glycoprotein is recognized by GP\g=a\HSRZ; and all three EZP glycoproteins separated by 1D-PAGE are recognized by R\g=a\HSPZ.These data add further support to the concept of cross-species zona pellucida glycoprotein antigenicity.