Sex sorting procedure is believed to induce changes in sperm that could be responsible for reducing the overall quality of sexed semen. The aim of this work was to verify by indirect immunofluorescence technique whether the process is responsible for capacitation-related changes in actin cytoskeleton polymerization and in protein tyrosine phosphorylation (PTP). We studied the distribution of separate sperm subpopulations through immune-reactive pattern in fresh (F) capacitated (Cap) and acrosome reacted (AR) sperm and compared these results with those observed in sexed spermatozoa. As for actin, three different patterns (A, B and C) were observed. A was mainly observed in F (A 91.8 ± 0.7%; B 5.8 ± 0.5%; C 2.4 ± 0.2%, mean ± SEM), B in Cap (A 21.3 ± 6.3%; B 70.4 ± 5.1%; C 8.3 ± 1.2%) and C in AR (A 5.1 ± 0.8%; B 24.5 ± 2.3%; C 70.5 ± 3.8%) sperm. In sexed sperm, the most expressed pattern was A (61.4 ± 8.6%), while B and C were 28.9 ± 8.6% and 5.8 ± 1%, respectively. No significant differences were observed between B and C patterns in sexed and F sperm, while A pattern was significantly different (p < 0.05) from that observed in F, Cap and AR sperm. As for PTP, we identified three patterns a, b and c. ‘‘a’’ pattern was typical of F cells (a 88.2 ± 3.7%; b 9.2 ± 3.1%; c 2.6 ± 1%), ‘‘b’’ was typical of Cap sperm (a 19.4 ± 5.4%; b 68.9 ± 5.8%; c 11.7 ± 0.4%) and ‘‘c’’ was mainly present in AR sperm (a 2.9 ± 0.7%; b 17.8 ± 2.7%; c 79.6 ± 2.1%). In sexed sperm (a 80.3 ± 3.3%; b 8.3 ± 3.3% and c 11.5 ± 2.8%) no significant differences in a and b patterns were found when compared with F sperm, while c was significantly higher (p < 0.05). We suggest that sex sorting procedure induces some changes in actin cytoskeleton polymerization that could represent a partial capacitative activation, while this kind of activation seems to be lower for PTP. Sexing increases acrosome reacted cells possibly due to mechanical damage rather to a direct stimulation of acrosome reaction.

Actin distribution and tyrosine phosphorylation in sex-sorted bull spermatozoa

SPINACI, MARCELLA;VALLORANI, CLAUDIA;GALEATI, GIOVANNA;TAMANINI, CARLO;BUCCI, DIEGO
2011

Abstract

Sex sorting procedure is believed to induce changes in sperm that could be responsible for reducing the overall quality of sexed semen. The aim of this work was to verify by indirect immunofluorescence technique whether the process is responsible for capacitation-related changes in actin cytoskeleton polymerization and in protein tyrosine phosphorylation (PTP). We studied the distribution of separate sperm subpopulations through immune-reactive pattern in fresh (F) capacitated (Cap) and acrosome reacted (AR) sperm and compared these results with those observed in sexed spermatozoa. As for actin, three different patterns (A, B and C) were observed. A was mainly observed in F (A 91.8 ± 0.7%; B 5.8 ± 0.5%; C 2.4 ± 0.2%, mean ± SEM), B in Cap (A 21.3 ± 6.3%; B 70.4 ± 5.1%; C 8.3 ± 1.2%) and C in AR (A 5.1 ± 0.8%; B 24.5 ± 2.3%; C 70.5 ± 3.8%) sperm. In sexed sperm, the most expressed pattern was A (61.4 ± 8.6%), while B and C were 28.9 ± 8.6% and 5.8 ± 1%, respectively. No significant differences were observed between B and C patterns in sexed and F sperm, while A pattern was significantly different (p < 0.05) from that observed in F, Cap and AR sperm. As for PTP, we identified three patterns a, b and c. ‘‘a’’ pattern was typical of F cells (a 88.2 ± 3.7%; b 9.2 ± 3.1%; c 2.6 ± 1%), ‘‘b’’ was typical of Cap sperm (a 19.4 ± 5.4%; b 68.9 ± 5.8%; c 11.7 ± 0.4%) and ‘‘c’’ was mainly present in AR sperm (a 2.9 ± 0.7%; b 17.8 ± 2.7%; c 79.6 ± 2.1%). In sexed sperm (a 80.3 ± 3.3%; b 8.3 ± 3.3% and c 11.5 ± 2.8%) no significant differences in a and b patterns were found when compared with F sperm, while c was significantly higher (p < 0.05). We suggest that sex sorting procedure induces some changes in actin cytoskeleton polymerization that could represent a partial capacitative activation, while this kind of activation seems to be lower for PTP. Sexing increases acrosome reacted cells possibly due to mechanical damage rather to a direct stimulation of acrosome reaction.
Reproduction in Domestic Animals
151
151
M Spinaci; C Vallorani; G Galeati; C Tamanini; J Rodriguez-Gil; D Bucci
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/11585/108701
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