2024-03-28T16:19:48Zhttp://buleria.unileon.es/oai/requestoai:buleria.unileon.es:10612/108292022-05-19T08:56:15Zcom_10612_17col_10612_18
2019-05-24T09:55:59Z
urn:hdl:10612/10829
Cleaved PARP‐1, an Apoptotic Marker, can be Detected in Ram Spermatozoa
Casao Gascón, Adriana
Mata Campuzano, María
Ordás, L.
Cebrián Pérez, José A.
Muiño Blanco, María Teresa
Martínez Pastor, Felipe
Biologia Celular
Veterinaria
P. 688-691
The presence of apoptotic features in spermatozoa has been related to lower quality and functional impairment. Members of the poly‐ADP‐ribose polymerases (PARP) familyare involved in both DNA repair and apoptosis, playing important roles in spermatogenesis. Poly‐ADP‐ribose polymerase can be cleaved by caspases, and the presence of its cleavage product (cPARP) in spermatozoa has been related to chromatin remodelling during spermatogenesis and to the activation of apoptotic pathways. There are no reports on immunodetection of cPARP in ram spermatozoa; thus, we have tested a commercially available antibody for this purpose. cPARP was microscopically detected in the acrosomal ridge of some spermatozoa (indirect immunofluorescence). A preliminary study was carried out by flow cytometry (direct immunofluorescence, FITC). Ram semen was extended in TALP and incubated for 4 h with apoptosis inducers staurosporine (10 μm) or betulinic acid (200 μm). Both inducers and incubation caused a significant increase in cPARP spermatozoa (0 h, control: 21.4±3.3%, inducers: 44.3±1.4%; 4 h, control: 44.3±2.4%, inducers: 53.3±1.4%). In a second experiment, we compared the sperm fractions after density gradient separation (pellet and interface). The pellet yielded a slightly lower proportion of cPARP spermatozoa (28.5±1.2% vs 36.2±2.0% in the interface; p < 0.001), and a 12‐h incubation increased cPARP similarly in both fractions (p < 0.001). cPARP seems to be an early marker of apoptosis in ram semen, although its presence in untreated samples was weakly related to worse quality (pellet/interface). We suggest to study the relationship of PARP and cPARP levels with between‐male differences on sperm fertility.
2019-05-24T09:55:59Z
2019-05-24T09:55:59Z
2019-05-24
info:eu-repo/semantics/article
Reproduction in Domestic Animals, 2015, vol. 50, n. 4
https://onlinelibrary.wiley.com/doi/full/10.1111/rda.12549
http://hdl.handle.net/10612/10829
John Wiley & Sons