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Regular Article |
a Department of Biochemistry and Molecular and Cell Biology, School of Veterinary Medicine, University of Zaragoza, 50013 Zaragoza, Spain
ABSTRACT
Ejaculated ram spermatozoa, freed from seminal plasma by a dextran/swim-up procedure and exposed to cold shock, were incubated with ram seminal plasma proteins and analyzed by fluorescence markers and scanning electron microscopy. Seminal plasma proteins bound to the sperm plasma membrane modified the functional characteristics of damaged spermatozoa, reproducing those of live cells. Scanning electron microscopy showed that the dramatic structural damage induced by cooling reverted after incubation with seminal plasma proteins. Assessment of membrane integrity by fluorescence markers also indicated a restoration of intact-membrane cells. This protein adsorption is a concentration-dependent process that induces cell surface restoration in relation to the amount of protein in the incubation medium. Fractionation of ram seminal plasma proteins by exclusion chromatography provided three fractions able to reverse the cold shock effect. Scanning electron microscopy also confirmed the high activity of one fraction, because approximately 50% of cold-shocked sperm plasma membrane surface was restored to its original appearance after incubation. Differences in composition between the three separated fractions mainly resulted from one major band of approximately 20 kDa, which must be responsible for recovering the sperm membrane permeability characteristic of a live cell.
First decision: 3 February 2000.
1 Supported by grants DGICYT AGF-97-0919 and DGA-48/99AV.
2 Correspondence: J.A. Cebrián Pérez, Departamento de Bioquímica y Biología Molecular y Celular, Facultad de Veterinaria, C/Miguel Servet, 177. 50013 Zaragoza, Spain. FAX: 34 976 761612; pcebri\|[aacute]\|n{at}posta.unizar.es
3 Current address: Servicio de Soporte a la Investigación Experimental, Universidad de Valencia, 46100 Burjassot, Valencia, Spain.
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