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Chlortetracycline fluorescence patterns and in vitro fertilisation of frozen-thawed boar spermatozoa incubated under various bicarbonate concentrations

Published online by Cambridge University Press:  26 September 2008

Lalantha R. Abeydeera ,
Hiroaki Funahashi ,
Nam-Hyung Kim  and
Billy N. Day
Lalantha R. Abeydeera
Affiliation:
Department of Animal Sciences, University of Missouri-Columbia, Columbia, Missouri, USA
Hiroaki Funahashi
Affiliation:
Department of Animal Sciences, University of Missouri-Columbia, Columbia, Missouri, USA
Nam-Hyung Kim
Affiliation:
Department of Animal Sciences, University of Missouri-Columbia, Columbia, Missouri, USA
Billy N. Day*
Affiliation:
Department of Animal Sciences, University of Missouri-Columbia, Columbia, Missouri, USA
*
Billy N. Day, University of Missouri-Columbia, Department of Animal Sciences, 159 Animal Sciences Research Center, Columbia, MO 65211, USA. Tel: +1(573)-882-7555. Fax: +1(573)-884-7827.
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Summary

Porcine oocyte-cumulus complexes were cultured in bovine serum albumin(BSA)-free North Carolina State University (NCSU)23 medium containing porcine follicular fluid (10%), cysteine (0.1 mg/ml)and hormonal supplements (eCG and hCG: 10IU/ml each) for 22h. They were then cultured in the same medium but without hormonal supplements for an additional 22h. After culture, cumulus cells were removed and oocytes were co-incubated with frozen-thawed ejaculated boar spermatozoa in tissue culture medium (TCM)199 containing caffeine (5mM), fetal calf serum (FCS; 10%) and varying concentrations (26–56 mM)of NaHCO3 for 9h(experiment 1). In experiment 2, chlortetracycline (CTC) was used to assess the functional state of spermatozoa incubated under different NaHCO3 concentrations. Experiment 3 examined the effect of FCS (1% and 10%)and NaHCO3 (26 and 46 mM) on fertilisation parameters. Compared with 26 mM, penetration rate was significantly higher (p<0.05)at 36–56mM NaHCO3. Polyspermy showed a similar pattern although no difference was observed between 26 and 36 mM. At 46mM NaHCO3, the mean number of spermatozoa (MNS) penetrated per oocyte increased significantly (p< 0.05). A significantly higher proportion of spermatozoa were capacitated and acrosome reacted at 46 and 56mM NaHCO3, respectively. The fertilisation medium containing 46mM NaHCO3 and 1% FCS showed a higher penetration rate (84%)with a relatively low incidence of polyspermy (39%). The results indicate that NaHCO3 stimulates capacitation and/or the acrosome reaction of boar spermatozoa in a dose-dependent manner and thus affects fertilisation parameters.

Keywords

Acrosome reactionBicarbonateCapacitationIn vitro fertilisationPig oocytes
Type
Article
Information
Zygote , Volume 5 , Issue 2 , May 1997 , pp. 117 - 125
Copyright
Copyright © Cambridge University Press 1997

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