Penetration of concanavalin-A-treated Chinese hamster oocytes by golden hamster spermatozoa in vitro, and chromosome analysis of hybrid 1-cell zygotes.

Abstract

Pretreatment of zona-free Chinese hamster (CH) oocytes with three kinds of lectin--concanavalin A (Con-A), phytohaemagglutinin-P (PHA) and wheat germ agglutinin (WGA)--was attempted in order to improve penetration by golden hamster (GH) spermatozoa in vitro. Con-A had no significant effect on penetration at 2 micrograms/ml, adequately facilitated oocyte-sperm fusion at 4 micrograms/ml, and caused excessive sperm binding and resultant severe polyspermy at 10 micrograms/ml. Neither PHA nor WGA had positive effects on sperm penetration at any concentrations (2-10 micrograms/ml) examined. Using the Con-A (4 micrograms/ml) pretreatment, high rates of interspecific fertilisation and subsequent chromosome analysis of hybrid 1-cell zygotes were achieved. Among 258 CH oocytes used, 212 (82.2%) were fertilised and 153 (72.2% of fertilised ova) developed to the first cleavage metaphase. Eventually, 132 CH-derived chromosome complements and 153 GH-derived ones were successfully karyoanalysed. Incidences of aneuploidy and structural anomaly were 3.1% and 2.3% in CH complements, and 1.4% and 6.5% in GH complements, respectively. These incidences were not significantly different from those obtained by intraspecific in vivo fertilisation, suggesting that our interspecific in vitro fertilisation system does not cause chromosome aberrations.