Structural and functional analysis of zona pellucida

• Main Author: Koothan, Thillai
• Published: University of Edinburgh 1994
• Subjects: 612.6
• Online Access: http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.653544

The ability of the zona pellucida to mediate the species specific recognition of spermatozoa is due to a glycoprotein constituent of this structure, known as ZP3. In view of its important role in the initiation of fertilization, ZP3 has been identified as a suitable candidate for the development of a contraceptive vaccine. In order to generate recombinant marmoset ZP3 for immunization purposes a full length 1.3 kb insert encoding for marmoset ZP3 was cloned downstream of the malE gene to generate a fusion protein containing marZP3 and a maltose binding domain. This protein was found to be generated in a soluble form in aqueous solutions and the antigenic integrity of this molecule was demonstrated by it capacity to be recognized by a monoclonal antibody raised against human ZP3. This full length fusion protein has been purified by anion exchange chromatography and used to generate polyclonal antibodies. In addition to these <I>in vitro</I> studies on the generation of a recombinant marZP3 for active immunization purposes, <I>in vivo</I> studies have also been performed on the induction of active immunity against porcine ZP3. These studies have demonstrated that an important side effect of induction of immunity against a heterologous ZP3 antigen involves the depletion of the primordial follicle pool. Moreover this pathology could be transferred to non-immunized recipients by the passive transfer of antibody. These data raise the important question as to whether a similar pathology would be induced if homologous zona antigens were used for the induction of immunity. As a result of the work descriebd in this thesis, this question can now be addressed in the marmoset monkey using recombinant marZP3 as antigen.