# FUNCTION OF SPERM MEMBRANE ANTIGENS IN FERTILIZATION

> **NIH NIH R01** · DUKE UNIVERSITY · 1994 · $139,563

## Abstract

The proposed research is aimed at understanding the molecular details of
mouse gamete interaction.  We have generated monoclonal anti-sperm
antibodies that have been used to identify proteins involved in two
distinct fertilization events:  the acrosome reaction and sperm-egg
membrane fusion.  In the present research plan, we will focus on sperm
interaction with the zona pellucida (ZP).

Several strategies will be used to identify the mouse sperm receptor(s) for
zp.  Monoclonal antibodies (mAbs) that block binding to zp will be sought
by immunizing mice with membrane fractions from epididymal sperm.  Three
different approaches are outlined for membrane preparation; this material
will be assayed for bioactivity, defined as the ability to compete with
sperm for zp binding.  Direct biochemical approaches using a) cross-linking
regents and b) RGD containing peptides to identify the sperm's zp
receptor(s) are also proposed.  Depending upon the complexity of the
preparations that result from these approaches, either conventional
biochemical or immuno-affinity techniques will be used to isolate the
receptor(s).

A second major aim is to test the hypothesis that interaction between
capacitated sperm and ZP3 results in receptor aggregation that itself is a
primary event in initiating the cascade terminating in the acrosome
reaction.  A variety of experimental approaches to examine receptor
aggregation and its possible physiological significance are proposed.  In
addition, bioactive membrane fractions, generated as part of the zp
receptor studies, will be characterized for activities known to be involved
in signal transduction, such as G proteins and endogenous protein kinases.
MAbs developed in the zp receptor studies will be examined for inhibitory
effects on AR triggering.  Two of our existing mAbs, M42 and M5 are initial
candidates to examine in detail.

Sperm from the proximal epididymis are incapable of fertilization.  We plan
to examine biochemical changes that occur in specific sperm components
during epididymal transit.  We have already found that the M42 antigen is
modified structurally in the epididymis; this modification coincides with
acquisition of function.  Co-culture of different epididymal sperm
populations with epididymal epithelia in primary culture will be used to
explore the biochemical maturation of relevant antigens in vitro.

## Key facts

- **NIH application ID:** 2197605
- **Project number:** 5R01HD018201-10
- **Recipient organization:** DUKE UNIVERSITY
- **SAM.gov UEI:** TP7EK8DZV6N5
- **Principal Investigator:** PATRICIA M SALING
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1994
- **Award amount:** $139,563
- **Award type:** 5
- **Project period:** 1983-04-01 → 1994-11-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/2197605

## Citation

> US National Institutes of Health, RePORTER application 2197605, FUNCTION OF SPERM MEMBRANE ANTIGENS IN FERTILIZATION (5R01HD018201-10). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/2197605. Licensed CC0.

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