The aim of the present study was to evaluate whether progesterone (P4) promotes sperm capacitation in buffalo. Frozen-thawed sperm were treated by swim-up to select only the motile population. Spermatozoa (N=1740, equally distributed into 6 groups) were incubated in the presence of: A) 0.1 µg of P4; B) 0.01 mM heparin and C) in medium lacking capacitating agents (control) for 1 and 2 hours. Sperm were exposed for 15 min to 60 g mL-1 of lysophosphatidylcholine, known to induce acrosome reaction only on capacitated spermatozoa. Spermatozoa were fixed in 37 % formaldheyde and double stained with Trypan blue and Giemsa, for acrosome evaluation by microscopic examination. The proportion of acrosome-reacted (AR) spermatozoa in each group was used to assess the efficiency of capacitation. Differences between groups were analyzed by Chi Square. The 1-hour treatment with both P4 and heparin significantly (P<0.01) increased the percentages of AR spermatozoa compared to the control (35.7 %, 23.2 % and 13.2 % respectively). Sperm incubation with P4 for 1 h resulted in significantly (P<0.01) higher incidence of AR compared to that with heparin, that is the capacitating agent currently used for in vitro fertilization (IVF). However, following 2 hours of incubation, both P4 and heparin gave similar percentages of AR sperm (35.6 vs 34.5 % respectively) that were significantly (P<0.01) higher than the control (13.3 %). It was demonstrated that P4 induces buffalo sperm capacitation in vitro and may be considered as an alternative capacitating agent for buffalo IVF.

Effect of progesterone on capacitation of buffalo (Bubalus bubalis) spermatozoa in vitro.

BOCCIA, LUCIA;ATTANASIO, LAURA;DE ROSA, ANNA;DI PALO, ROSSELLA;GASPARRINI, BIANCA
2006

Abstract

The aim of the present study was to evaluate whether progesterone (P4) promotes sperm capacitation in buffalo. Frozen-thawed sperm were treated by swim-up to select only the motile population. Spermatozoa (N=1740, equally distributed into 6 groups) were incubated in the presence of: A) 0.1 µg of P4; B) 0.01 mM heparin and C) in medium lacking capacitating agents (control) for 1 and 2 hours. Sperm were exposed for 15 min to 60 g mL-1 of lysophosphatidylcholine, known to induce acrosome reaction only on capacitated spermatozoa. Spermatozoa were fixed in 37 % formaldheyde and double stained with Trypan blue and Giemsa, for acrosome evaluation by microscopic examination. The proportion of acrosome-reacted (AR) spermatozoa in each group was used to assess the efficiency of capacitation. Differences between groups were analyzed by Chi Square. The 1-hour treatment with both P4 and heparin significantly (P<0.01) increased the percentages of AR spermatozoa compared to the control (35.7 %, 23.2 % and 13.2 % respectively). Sperm incubation with P4 for 1 h resulted in significantly (P<0.01) higher incidence of AR compared to that with heparin, that is the capacitating agent currently used for in vitro fertilization (IVF). However, following 2 hours of incubation, both P4 and heparin gave similar percentages of AR sperm (35.6 vs 34.5 % respectively) that were significantly (P<0.01) higher than the control (13.3 %). It was demonstrated that P4 induces buffalo sperm capacitation in vitro and may be considered as an alternative capacitating agent for buffalo IVF.
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/11588/105352
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