Studies on the fertilization of mouse eggs <I>in vitro</I>:II. Effects of <I>in vitro</I> pre-incubation of spermatozoa on time of sperm penetration of mouse eggs <I>in vitro</I>
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Capacitation <I>in vitro</I> of mouse spermatozoa was studied by examining the time of <I>in vitro</I> penetration of eggs by previously incubated sperm.<BR>Sperms recovered from cauda epididymis of adult males of ICR-JCL strain were suspended in a chemically defined medium (Krebs-Ringer-Bicarbonate solution supplemented with glucose, Na-pyruvate, bovine albumin and antibiotics) and incubated at 37°C under 5% CO<SUB>2</SUB> in air for various period before they were used for insemination.<BR>When sperms incubated for 2 hr were used for insemination, the rate of sperm penetration through the zona pellucida, as examined at 1/2, 1, 2, 3, or 4 hr after insemination, was 57.1, 100.0, 100.0, 98.9 or 98.7% respectively, in comparison with 0, 22.8, 96.4, 93.9 or 90.9% for fresh epididymal sperm (Table 1 and Fig. 1). In the former group, 96% of eggs examined at 1 hr after insemination were at telophase of second maturation division with enlarged sperm head(s) in vitellus, while in the latter group, only 10.9% of eggs were found to be fertilized at this time. It is, thus, clear that in-cubated spermatozoa penetrate the eggs sooner than the fresh ones. The mean number of penetrating sperm per penetrated egg was also higher (3.1) in eggs inseminated with incubated sperm than that (1.6) in eggs with fresh sperm (Fig. 2). <BR>When the eggs were inseminated with spermatozoa which had been incubated for 37, 15, 30, 60 or 120 minutes, 24.1, 70.1, 93.9, 98.7 or 100% of eggs were found to be penetrated, and 5.7, 46.7, . 73.7, 89.5 or 96.0% of them were found to be fertilized at 1hr after insemination (Table 2). This indicates that the change occurred in sperms during incubation is of a progressive nature and is a fairly rapid one, virtually completing within 1 hr after the start of incubation.<BR>These results seem to show that mouse epididymal spermatozoa can be capacitated <I>in vitro</I> in a chemically defined medium without the presence of female reproductive tissue fluid. This will favor the view that capacitation, at least in mouse, is dependent on some appropriate physico-chemical environment rather than on some specific substances from the female reproductive tissues.
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