Pannexin channels increase propidium iodide permeability in frozen-thawed dog spermatozoa
Author
dc.contributor.author
Torres, J. L.
Author
dc.contributor.author
Palomino, J.
Author
dc.contributor.author
Moreno, R. D.
Author
dc.contributor.author
Reyes Solovera, Mónica de los
Admission date
dc.date.accessioned
2018-12-20T14:53:41Z
Available date
dc.date.available
2018-12-20T14:53:41Z
Publication date
dc.date.issued
2017
Cita de ítem
dc.identifier.citation
Reproduction, Fertility and Development, 2017, 29, 2269–2276
Identifier
dc.identifier.issn
14485990
Identifier
dc.identifier.issn
10313613
Identifier
dc.identifier.other
10.1071/RD16267
Identifier
dc.identifier.uri
https://repositorio.uchile.cl/handle/2250/157360
Abstract
dc.description.abstract
Pannexins (Panx) are proteins that form functional single membrane channels, but they have not yet been
described in dogs. The aim of the present study was to detect Panx1, Panx2 and Panx3 in frozen–thawed dog spermatozoa
using flow cytometry and immunofluorescence analyses, evaluating the relationship of these proteins with propidium
iodide (PI) in frozen–thawed spermatozoa. Fresh and frozen–thawed dog spermatozoa from eight dogs were preincubated
with 3 mM PI with or without 15 mM carbenoxolone (CBX) or 1 mM probenecid (PBD), two Panx channel inhibitors, and
then incubated with rabbit anti-Panx1, anti-Panx2 and anti-Panx3 antibodies (1 : 200). Panx immunolocalisation was
assessed by fluorescence microscopy. Flow cytometry data were evaluated by analysis of variance. All three Panx proteins
were found in dog spermatozoa: Panx1 was mostly localised to the acrosomal and equatorial segment, Panx2 was found in
the posterior region of the head and tail and Panx3 was localised to the equatorial and posterior head segment. The
percentage of PI-positive cells determined by flow cytometry was reduced (P , 0.05) in the presence of Panx inhibitors.
These results show that Panx proteins are present in dog spermatozoa and increase PI permeability in frozen–thawed dog
sperm, suggesting that the percentage of PI-positive spermatozoa used as an indicator of non-viable cells may lead to
overestimation of non-viable cells.