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    Título
    Cleaved PARP‐1, an Apoptotic Marker, can be Detected in Ram Spermatozoa
    Autor
    Casao Gascón, Adriana
    Mata Campuzano, María
    Ordás, L.
    Cebrián Pérez, José A.
    Muiño Blanco, María Teresa
    Martínez Pastor, FelipeAutoridad BuleriaORCID
    Facultad/Centro
    Facultad de Ciencias Biologicas y Ambientales
    Área de conocimiento
    Biologia Celular
    Título de la revista
    Reproduction in Domestic Animals
    Datos de la obra
    Reproduction in Domestic Animals, 2015, vol. 50, n. 4
    Editor
    John Wiley & Sons
    Fecha
    2015-08
    Abstract
    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.
    Materia
    Veterinaria
    Palabras clave
    Polimerasas
    Esperma
    Ganado ovino
    Peer review
    SI
    URI
    http://hdl.handle.net/10612/10829
    Versión del editor
    https://onlinelibrary.wiley.com/doi/full/10.1111/rda.12549
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