Evaluation of the expression of sperm proteins in normozoospermic and asthenozoospermic sperm samples using monoclonal antibodies, flow cytometry and fluorescence microscopy

0463171 - BTÚ 2017 US eng C - Konferenční příspěvek (zahraniční konf.)
Děd, Lukáš - Čapková, Jana - Kubátová, Alena - Teplá, O. - Pěknicová, Jana
Evaluation of the expression of sperm proteins in normozoospermic and asthenozoospermic sperm samples using monoclonal antibodies, flow cytometry and fluorescence microscopy.
Abstract book of the 48th Annual Meeting of the Society for the Study of Reproduction. San Juan: Society for the Study of Reproduction, 2015.
[48th Annual Meeting of the Society for the Study of reproduction. San Juan (US), 18.06.2015-22.06.2015]
Grant CEP: GA ČR(CZ) GA14-05547S; GA MŠMT(CZ) ED1.1.00/02.0109; GA ČR(CZ) GA P503/12/1834
Klíčová slova: asthenozoospermia * normozoospermia * monoclonal antibody * flow cytometry
Kód oboru RIV: EC - Imunologie

Asthenozoospermia is one of the main seminal pathologies underlying male infertility. In our study we aimed to evaluate the ability of specific antibodies to detect the differential expression of selected protein markers by fluorescent microscopy and flow cytometry techniques. Therefore we analyzed sperm samples from 30 men with normal spermiograms and 30 men with asthenozoospermia (<40% motile spermatozoa) by the panel of our diagnostic anti-human sperm (Hs) antibodies. These antibodies were prepared in our laboratory and some of them are used in clinical practice as a tool for the differential diagnosis of various sperm pathologies. Both fluorescent microscopy and flow cytometry analysis revealed quantitative differences in the protein abundances between normozoospermic and asthenozoospermic sperm samples, namely, in GAPDHS, evaluated with Hs-8 MoAb, VCP (valosin-containing protein), evaluated with Hs-14 MoAb, and ATP synthase (cAMP-dependent protein kinase II, PRKAR2A), evaluated with MoAb Hs-36. On the other hand no statistically significant differences were found in the expression of the sperm surface protein clusterin, evaluated with Hs-3 MoAb, and semenogelin, evaluated with Hs-9 MoAb. From the methodological point of view, we observed very high correlation between the data obtained by fluorescent microscopy and flow cytometry techniques. From the clinical point of view, we observed the strong association of the low sperm motility in the sample not only with the expression of proteins playing a important role in sperm energy metabolism (expected), but also with the expression of all tested intra-acrosomal proteins. These findings further demonstrate asthenozoospermia as a complex semen disorder frequently associated with other semen pathologies which are not diagnosed by basic semen analysis and the possibility to use monoclonal antibodies as a tool for diagnosis of protein associated sperm pathologies in the semen with the low sperm motility.
Trvalý link: http://hdl.handle.net/11104/0262874