Method of Determining Sperm Capacitation
AssigneesCornell University, The Trustees of the University of Pennsylvania
InventorsAlexander J. Travis, Gregory S. Kopf
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Abstract
This invention describes unique patterns of distribution of ganglioside GM1 in non-capacitated sperm and demonstrates that the pattern of distribution of GM1 undergoes changes that can be correlated with the process of capacitation and/or with acrosomal exocytosis. Accordingly, the present invention discloses a method for determining the ability of sperm to respond to capacitation and/or acrosomal exocytosis stimuli. The method comprises determination of distribution pattern for GM1. The method can be used for both diagnostic and predictive purposes when assessing male reproductive fitness, and can also be used to assess the effects of any agent or environment on sperm including cryoprotective agents and protocols, and contraceptive agents.
Claims20
A method of determining the effect of a test agent on the ability of sperm to respond to one or more stimuli associated with capacitation and/or acrosomal exocytosis comprising the steps of:
- a) obtaining sperm which have been exposed to the test agent;
- b) determining a GM1 distribution pattern in the sperm from step a) comprising;
- i) exposing the sperm to the one or more stimuli;
- ii) fixing the sperm in a fixative comprising paraformaldehyde, glutaraldehyde or combinations thereof;
- iii) staining the fixed sperm to determine the GM1 distribution pattern in the fixed test sperm which had been exposed to the test agent
- c) obtaining a GM1 distribution pattern of a fixed control sperm sample which has not been exposed to the test agent, but which has been exposed to the same one or more stimuli, fixed in the same fixative and stained in the same manner as the sperm in step b);
- d) comparing the GM1 distribution pattern of the fixed test sperm and the fixed control sperm;
- wherein a difference between the GM1 distribution patterns of the fixed test sperm and fixed control sperm is indicative of an effect of the test agent on the ability of sperm to respond to one or more stimuli associated with capacitation and/or acrosomal exocytosis.
A method of determining the effect of a test agent on the ability of sperm to respond to one or more stimuli associated with capacitation and/or acrosomal exocytosis comprising the steps of:
- a) obtaining sperm from an individual who has been exposed to the test agent;
- b) determining a GM1 distribution pattern in the sperm from step a) comprising;
- i) exposing the sperm to the one or more stimuli;
- ii) fixing the sperm in a fixative comprising paraformaldehyde, glutaraldehyde or combinations thereof;
- iii) staining the fixed sperm to determine the GM1 distribution pattern in the fixed test sperm;
- c) obtaining a GM1 distribution pattern of a fixed control sperm sample which is obtained from an individual who has not been exposed to the test agent, but which has been exposed to the same one or more stimuli, fixed in the same fixative and stained in the same manner as the sperm in step b);
- d) comparing the GM1 distribution pattern of the fixed test sperm and the fixed control sperm;
- wherein a difference between the GM1 distribution patterns of the fixed test sperm and fixed control sperm is indicative of an effect of the test agent on the ability of sperm to respond to one or more stimuli associated with capacitation and/or acrosomal exocytosis.
Dependencies are inferred from the claim text.