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dc.contributor.author
Bedford Guaus, Sylvia J.
dc.contributor.author
Mc Partlin, L. A.
dc.contributor.author
Xie, J.
dc.contributor.author
Westmiller, S. L.
dc.contributor.author
Buffone, Mariano Gabriel
dc.contributor.author
Roberson, M. S.
dc.date.available
2016-12-28T17:03:11Z
dc.date.issued
2011-07
dc.identifier.citation
Bedford Guaus, Sylvia J.; Mc Partlin, L. A. ; Xie, J. ; Westmiller, S. L. ; Buffone, Mariano Gabriel; et al.; Molecular cloning and characterization of Phospholipase C Zeta in equine sperm and testis reveals species-specific differences in expression of catalytically active protein; Society For The Study Of Reproduction; Biology Of Reproduction; 85; 1; 7-2011; 78-88
dc.identifier.issn
0006-3363
dc.identifier.uri
http://hdl.handle.net/11336/10476
dc.description.abstract
Oocyte activation at fertilization is brought about by the testis-specific phospholipase C zeta (PLCZ), owing to its ability to induce oscillations in intracellular Ca(2+) concentration ([Ca(2+)](i)). Whereas this is a highly conserved mechanism among mammals, important species-specific differences in PLCZ sequence, activity, and expression have been reported. Thus, the objectives of this research were to clone and characterize the intracellular Ca(2+)-releasing activity and expression of equine PLCZ in sperm and testis. Molecular cloning of equine PLCZ yielded a 1914-bp sequence that translated into a protein of the appropriate size (~73 kDa), as detected with an anti-PLCZ-specific antibody. Microinjection of 1 μg/μl of equine PLCZ cRNA supported [Ca(2+)](i) oscillations in murine oocytes that were of a higher relative frequency than those generated by an equivalent concentration of murine Plcz cRNA. Immunofluorescence revealed expression of PLCZ over the acrosome, equatorial segment, and head-midpiece junction; unexpectedly, PLCZ also localized to the principal piece of the flagellum in all epididymal, uncapacitated, and capacitated sperm. Immunostaining over the acrosome was abrogated after induction of acrosomal exocytosis. Moreover, injection of either sperm heads or tails into mouse oocytes showed that PLCZ in both fractions is catalytically active. Immunohistochemistry on equine testis revealed expression as early as the round spermatid stage, and injection of these cells supported [Ca(2+)](i) oscillations in oocytes. In summary, we report that equine PLCZ displays higher intrinsic intracellular Ca(2+)-releasing activity than murine PLCZ and that catalytically active protein is expressed in round spermatids as well as the sperm flagellum, emphasizing important species-specific differences. Moreover, some of these results may suggest potential novel roles for PLCZ in sperm physiology.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Society For The Study Of Reproduction
dc.rights
info:eu-repo/semantics/openAccess
dc.rights.uri
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.subject
Sperm
dc.subject
Fertilization
dc.subject
Oocyte
dc.subject
Plczeta
dc.subject.classification
Biología Reproductiva
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Ciencias Biológicas
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CIENCIAS NATURALES Y EXACTAS
dc.title
Molecular cloning and characterization of Phospholipase C Zeta in equine sperm and testis reveals species-specific differences in expression of catalytically active protein
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2016-12-27T20:13:04Z
dc.identifier.eissn
1529-7268
dc.journal.volume
85
dc.journal.number
1
dc.journal.pagination
78-88
dc.journal.pais
Estados Unidos
dc.journal.ciudad
Nueva York
dc.description.fil
Fil: Bedford Guaus, Sylvia J.. Cornell University; Estados Unidos
dc.description.fil
Fil: Mc Partlin, L. A. . Cornell University; Estados Unidos
dc.description.fil
Fil: Xie, J. . Cornell University; Estados Unidos
dc.description.fil
Fil: Westmiller, S. L. . Cornell University; Estados Unidos
dc.description.fil
Fil: Buffone, Mariano Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina
dc.description.fil
Fil: Roberson, M. S.. Cornell University; Estados Unidos
dc.journal.title
Biology Of Reproduction
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/http://www.biolreprod.org/content/85/1/78.long
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