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D. B. MORTON

Summary.

Acrosin and hyaluronidase have been localized in the acrosomal region of ram spermatozoa using specific antibodies raised against the highly purified enzymes. Hyaluronidase staining was denser at the periphery of the sperm head, whereas acrosin staining was denser in the equatorial region and appeared to be bound to the inner acrosomal membrane.

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B. D. BAVISTER and D. B. MORTON

A fraction that stimulates hamster sperm motility can be obtained from human serum by Sephadex gel filtration (Morton & Bavister, 1974). The protein-rich fractions potentiate this activity and effectively induce the acrosome reaction. Here, we describe attempts to determine the type of protein that may be involved.

The methods used to prepare the motility-stimulating fraction from human serum and for assessing its activity were as previously described (Morton & Bavister, 1974). To help identify which serum component induced the acrosome reaction, serum proteins were chromatographically separated on Sephadex G-150 (5 ml pooled human serum fractionated on 130 ml bed volume G-150 in a 1·5×90 cm column, fraction size 1·6 ml, flow rate 10 ml/hr). Protein was monitored by absorption at 280 nm with an LKB Uvicord II (see Text-fig.

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D. B. MORTON and B. D. BAVISTER

Efficient capacitation of hamster spermatozoa can be induced by follicular fluid (Barros & Austin, 1967; Yanagimachi, 1969a, b) and blood sera of several species (Barros & Garavagno, 1970; Yanagimachi, 1970a; Talbot, Franklin & Fussell, 1974). Two components are involved: one is dialysable, heat-stable and stimulates sperm motility, while the other is non-dialysable, heat-labile and induces the acrosome reaction (Yanagimachi, 1969a). Our understanding of capacitation and the acrosome reaction might be enhanced if the nature and mode of action of these factors were known. Blood serum is an easily available source of both factors and this preliminary report describes the recovery of the sperm motility-stimulating activity from human serum.

Fresh human serum was heated to 56°C for 30 min to destroy unidentified toxic factor(s) (Yanagimachi, 1970a). The serum was chromatographed on Sephadex G-25 (medium grade) and equilibrated in Tyrode's solution supplemented with 0·33 mm-sodium pyruvate, 0·01 mg phenol red/ml,

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D. B. MORTON and T. D. GLOVER

Summary.

Rabbit does were killed at intervals between 2 min and 90 hr after mating and spermatozoa were flushed from the genital tract and counted. A technique is described which facilitates the counting of small numbers of spermatozoa with reasonable accuracy. Evidence was found to suggest that the rabbit's cervix functions as a sperm reservoir and factors influencing the formation of the reservoir and transcervical migration are discussed. It was noted that spermatozoa reached the oviduct within 30 min of coitus.

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D. B. MORTON and T. D. GLOVER

Summary.

Ovariectomized oestrogen-treated rabbits were inseminated with semen of variable sperm density and volume. Does were killed 10 hr after insemination and the number of spermatozoa in the vagina, cervix, uterus and oviduct was determined. It was found that sperm counts from all regions of the genital tract were directly related to the total number of spermatozoa in the inseminate. When the number of spermatozoa in the inseminate was held constant, more spermatozoa were recovered from the cervix and uterus with the smaller inseminate volumes. All regional counts were positively correlated with one another. Proportionately fewer spermatozoa were recovered from the genital tract when the inseminate contained larger sperm numbers.

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H. Morton, C. D. Nancarrow, R. J. Scaramuzzi, B. M. Evison and G. J. A. Clunie

Summary. The rosette inhibition test, an established test for determining the immunosuppressive potential of antilymphocyte serum, has been applied to the serum of sheep after mating. The rosette inhibition titre was much higher (12–26) in 7 sheep which were fertilized and remained pregnant for up to 21 days than in 5 sterile ewes mated with intact rams (8–10). The difference was apparent by 24 h after mating. One ewe had high titres for 6 days after mating but these then dropped and she returned to oestrus; early embryonic loss was suspected. Another ewe which returned to oestrus had consistently low titres. The results indicate that the rosette inhibition test can be used to detect fertilization, early embryonic death and continued pregnancy in sheep.