. Author manuscript; available in PMC: 2016 Oct 11.

Published in final edited form as: Mol Reprod Dev. 2010 Feb;77(2):136–148. doi: 10.1002/mrd.21114

TABLE 2.

Fresh Bovine Sperm Capacitated by Incubating With Ouabain or Anti-ATP1A4 Developed Into Pronuclei Following Penetration of the Zona Pellucida of Bovine Oocytes In Vitro

Group	No. of oocytes	Percentage of oocytes with two pronuclei (fertilization rate)
Negative	51	23.5.0^a
Heparin	58	70.7.6^b
Ouabain	57	71.9^b
Anti-ATP1A4	58	58.6^c
Pre-absorbed	51	21.5^a
Pre-immune	51	19.6^a

Sperm preparations were incubated with either Sp-TALP alone (negative), heparin (positive control for capacitation), ouabain, anti-ATP1A4, anti-ATP1A4 pre-absorbed to its blocking peptide or pre-immune serum for 4 hr. After incubation, sperm were washed in Sp-TALPH and resuspended in final fertilization medium, and used for inseminating in vitro-matured bovine oocytes. After 6 hr of sperm–oocyte coincubation, presumptive zygotes were cumulus-dissociated and cultured in SOF for an additional 12 hr for pronuclei development. These oocytes were fixed in acetic acid alcohol (1:3) for 24 hr and stained with 1% Orcein in glacial acetic acid and examined microscopically (400×; phase 2) for the demonstration of pronuclei.

Values without a common superscript (a–c) differed (P <0.05).