The possibility to control the sex of mammalian offspring raises great interest in the breeding technologies field and has considerable economic value in the livestock industry. Recent advances in cellular methodologies have led to the development of novel technologies to differentiate and separate X- and Y-chromosome-bearing sperm, although the only effective method for routine use is fluorescence-activated cell sorting using flow cytometry (1). Actually, sperm sorting is based on differential DNA content of X and Y chromosome-bearing sperm cells, which is approximately 4.2% in bovine. However, the reduced pregnancy success rates obtained by using sexed sperm by flow cytometry (2) and the potential hazards deriving by DNA-based separation techniques, which requires DNA staining and UV exposure, are relevant reasons to research alternative sorting methods. Over the years, many theoretical differences have been suggested to differentiate X- and Y-chromosome bearing sperm cells, including size, weight, density, swimming speed, electrical surface charges, surface macromolecular proteins. Nevertheless, the efforts to separate these two populations on the basis of chemical/physical properties have been unsuccessful, principally because the mean differences are narrow and the population distributions overlap considerably (3). Here, we report our proteomic investigations addressed to biochemical characterization of sexed bovine semen. The present study, aimed to indicate the more relevant proteome differences in order to differentiate X- and Y-chromosome bearing sperm cells, was conducted by using a comparative proteomic approach by nLC-MS/MS analysis. 17 proteins were found differentially expressed. 9 Differential proteins may be directly associated with the main components of cytoskeletal structures of sperm flagellum, as the axoneme, outer dense fibers and fibrous sheath, while 4 proteins are glycolitic enzymes and other 2 proteins may be associated to energetic metabolism. Overall the results provide a base to a better comprehension of structural differences between X- and Y-chromosome bearing sperm cells and may are useful to future research addressed to improve the actual sorting method.

Comparative analysis of bovine sexed sperm cells by nLC-MS/MS / M. De Canio, A. Soggiu, C. Piras, L. Bonizzi, A. Galli, A. Urbani, P. Roncada - In: 8th annual national conference : abstract volume / [a cura di] Italian Proteomics Association. - Padova : EdiSES, 2013 Jun. - ISBN 978-88-7959-775-3. - pp. 104-104 (( Intervento presentato al 8. convegno Annual national conference tenutosi a Padova nel 2013.

Comparative analysis of bovine sexed sperm cells by nLC-MS/MS

M. DE CANIO
Primo
;
A. Soggiu
Secondo
;
C. Piras;L. Bonizzi;
2013-06

Abstract

The possibility to control the sex of mammalian offspring raises great interest in the breeding technologies field and has considerable economic value in the livestock industry. Recent advances in cellular methodologies have led to the development of novel technologies to differentiate and separate X- and Y-chromosome-bearing sperm, although the only effective method for routine use is fluorescence-activated cell sorting using flow cytometry (1). Actually, sperm sorting is based on differential DNA content of X and Y chromosome-bearing sperm cells, which is approximately 4.2% in bovine. However, the reduced pregnancy success rates obtained by using sexed sperm by flow cytometry (2) and the potential hazards deriving by DNA-based separation techniques, which requires DNA staining and UV exposure, are relevant reasons to research alternative sorting methods. Over the years, many theoretical differences have been suggested to differentiate X- and Y-chromosome bearing sperm cells, including size, weight, density, swimming speed, electrical surface charges, surface macromolecular proteins. Nevertheless, the efforts to separate these two populations on the basis of chemical/physical properties have been unsuccessful, principally because the mean differences are narrow and the population distributions overlap considerably (3). Here, we report our proteomic investigations addressed to biochemical characterization of sexed bovine semen. The present study, aimed to indicate the more relevant proteome differences in order to differentiate X- and Y-chromosome bearing sperm cells, was conducted by using a comparative proteomic approach by nLC-MS/MS analysis. 17 proteins were found differentially expressed. 9 Differential proteins may be directly associated with the main components of cytoskeletal structures of sperm flagellum, as the axoneme, outer dense fibers and fibrous sheath, while 4 proteins are glycolitic enzymes and other 2 proteins may be associated to energetic metabolism. Overall the results provide a base to a better comprehension of structural differences between X- and Y-chromosome bearing sperm cells and may are useful to future research addressed to improve the actual sorting method.
Bovine sexed sperm ; proteomics ; shotgun-MS
Settore VET/05 - Malattie Infettive degli Animali Domestici
Italian Proteomics Association
ItPA
Book Part (author)
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/2434/221878
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