Evaluation of acrosome reaction by flow cytometry.

0482716 - BTÚ 2018 eng K - Conference Paper (Czech conference)
Dostálová, Pavla - Dvořáková-Hortová, Kateřina
Evaluation of acrosome reaction by flow cytometry.
[International Conference Analytical Cytometry IX. Praha (CZ), 14.10.2017-17.10.2017]
R&D Projects: GA ČR(CZ) GA 14-05547S; GA MŠMT(CZ) ED1.1.00/02.0109
Institutional support: RVO:86652036
Keywords : flow cytometry * sperm capacitation * EGFP
OECD category: Reproductive biology (medical aspects to be 3)

Ejaculated sperm are still not able to fertilize the egg. They first have to undergo a maturation process called capacitation. Only capacitated sperm can undergo the acrosome reaction and fertilize the egg. It is matter of debate where the sperm undergo the acrosome reaction, whether on the zona pellucida of the egg or whether earlier during their passage through the female reproductive tract or cumulus cells. Several detection methods for assessing the sperm maturation status have already been described, including tyrosine phosphorylation, chlortetracycline assay, PNA staining etc. All these methods require fixation of sperm. We are of possession of a transgenic mouse model that express EGFP under the acrosomal promotor and, as a result, the EGFP protein starts to be expressed in spermatids and localizes inside the acrosome in sperm. The loss of EGFP signal in the sperm acrosomal region indicates that sperm have undergone the acrosome reaction. This EGFP-sperm model is suitable for assessing the status of acrosome also on living sperm. Combining this model with flow cytometry is a great and strong tool for evaluating the impact of various substances such as pollutants or different inductors of acrosome reaction (progesterone, zona pellucida and calcium ionophore) on course of acrosome reaction.
Permanent Link: http://hdl.handle.net/11104/0278125