Publications by authors named "Mirjana Martic"

2 Publications

  • Page 1 of 1

Recombinant human zona pellucida proteins ZP1, ZP2 and ZP3 co-expressed in a human cell line.

Asian J Androl 2004 Mar;6(1):3-13

Department of Obstetrics and Gynaecology, University of Melbourne, 320 Victoria Parade, East Melbourne 3002 VIC, Australia.

Aim: To produce biologically active recombinant human (rh) ZP proteins in a human cell for use in sperm function tests.

Methods: The human embryonic kidney cell line 293T was employed to produce rhZP1, rhZP2 and rhZP3 proteins individually and together by co-expression. Presence of these proteins in the culture medium and cell lysate was assessed by Western blotting analysis. The effect of the recombinant proteins on the human AR was assessed.

Results: RhZP2 and rhZP3 were secreted into the culture medium, whereas rhZP1 was found only in the cell lysate. Interestingly, when all zona pellucida proteins were co-expressed in the same cells, rhZP1 was also secreted into the culture medium. However, despite the presence of all three ZP proteins in sufficient concentration and evidence of heavy glycosylation on gel electrophoresis, biological activity to induce the AR was not observed.

Conclusion: RhZP1, rhZP2 and rhZP3 were successfully expressed in the human embryonic kidney cell line 293T. It appears that an interaction amongst these proteins may be required for release of rhZP1 from the cell. Although this approach is not satisfactory for producing active human ZP proteins, it makes a significant contribution to the understanding of the structural and functional characteristics of the ZP proteins.
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March 2004

Phorbol myristate acetate induces ruffling of the acrosome of human sperm.

Fertil Steril 2002 Jul;78(1):128-36

Department of Obstetrics and Gynaecology, University of Melbourne, Melbourne, Royal Women's Hospital, Victoria, Australia.

Objective: To determine the effect of phorbol myristate acetate (PMA) on human acrosome morphology and the acrosome reaction.

Design: Controlled experiments on sperm and unfertilized oocytes from volunteers.

Setting: Academic research and teaching tertiary hospital.

Patient(s): Sperm samples were from normospermic men and unfertilized oocytes from IVF patients.

Main Outcome Measure(s): Acrosome morphology was assessed by using transmission and scanning electron microscopy. The acrosome reaction was assessed by using fluorescein-labeled Pisum sativum agglutinin.

Result(s): PMA induced acrosome ruffling, indicated by a marked wavy appearance. A significant correlation was found between PMA-induced ruffling and PMA enhancement of the zona pellucida-induced acrosome reaction. Protein kinase C inhibitors bisindolylmalemide I and sangivamycin had no effect on PMA-induced acrosomal ruffling, but actin polymerization inhibitors cytochalasin B and cytochalasin D significantly decreased PMA-induced acrosomal ruffling. In contrast, bisindolylmalemide I, sangivamycin, cytochalasin B, and cytochalasin D significantly decreased both the zona pellucida-induced acrosome reaction and the PMA enhancement of the zona pellucida-induced acrosome reaction.

Conclusion(s): PMA-induced acrosomal ruffling involves actin polymerization, possibly independent of conventional protein kinase C. Acrosomal ruffling is involved in the PMA augmentation of the zona pellucida-induced acrosome reaction.
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http://dx.doi.org/10.1016/s0015-0282(02)03166-7DOI Listing
July 2002