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Summary. Microspheres of various materials and diameters were transferred microsurgically to the rat oviduct on Day 1 of pregnancy and autopsies were done at various times thereafter up to Day 10 to assess the recovery and segmental distribution of microspheres and eggs in the genital tract or the viability of embryos. The number and distribution of eggs in the treated and control sides after unilateral transfers were not different on Day 4 and 5 and the number of embryos implanted on Day 10 was not significantly affected after bilateral transfers.

The segmental distribution of eggs and starch microspheres within the oviduct on Days 2, 3 or 4 showed that both are transported partly intermingled from ampulla to uterus. When microspheres of poly(dl-lactide-co-glycolide), starch, dextran or dextran blue were transferred, their distribution in the genital tract in the morning of Day 5 showed that poly(dl-lactide-co-glycolide) and dextran microspheres stayed longer in the oviduct while starch and dextran blue microspheres were transported to the uterus at the same time as the eggs. Transfer of starch microspheres of 40–60 μm to one oviduct and 180–200 μm in diameter to the opposite oviduct showed that distribution on one side was nearly identical to that of the other side from Days 2 to 5.

We conclude that the behaviour of synthetic surrogate ova in rats differs from that in rabbits. The rat oviduct does not change the rate of transport of native eggs following transfer of synthetic surrogate ova. Also, in the rat the composition of the surrogates has greater influence than their size on their time of passage to the uterus. Some surrogates mimic quite well the oviducal transport of embryos and can be used therefore to study this process in species in which the eggs are not coated with additional layers after ovulation.

Keywords: rat; oviduct; embryo transport; egg surrogate; microsphere

Summary. Starch or dextran blue microspheres were transferred microsurgically to the infundibulum of the oviduct on Days 1, 2, or 3 of pregnancy of control and oestradiol-treated rats. The animals were killed a few hours to several days after transfer to assess the number and distribution of ova and microspheres in the tract.

After transfer on Day 1 of pregnancy, microspheres and eggs crossed the ampullary-isthmic junction (AIJ) 18 h after ovulation. After transfer on Day 2 of pregnancy, more than 50% of microspheres were retained in the ampulla, indicating that the AIJ changes again 34 h after ovulation. Treatment with oestradiol did not advance the passage of eggs or microspheres across the AIJ but caused accelerated transport through the isthmus as soon as the eggs or microspheres reached this segment. Dextran blue microspheres were seen to move back and forth in the isthmus of control anaesthetized rats at a frequency of 5–6 times/min. Between 7 and 20 h after treatment with oestradiol the frequency of these movements was significantly augmented, indicating that increased frequency of contractions of the smooth muscle of the isthmus precedes and accompanies accelerated transport of ova through this segment.

Keywords: rat; oviduct; embryo transport; oestradiol; microspheres

Summary.

The isoelectric points of washed spermatozoa from intact boars and from boars after removal of the seminal vesicles were determined using isoelectric focusing on natural pH gradients. Normal boar spermatozoa focused at a higher pH than spermatozoa from boars without seminal vesicles. The isoelectric point of the latter was increased to a value approaching normal by preincubation in normal seminal plasma. This indicates that seminal plasma alters the membrane surface charge of boar spermatozoa on ejaculation.

Summary.

Seminal plasma basic proteins were labelled with ¹³¹I. The efficiency of the labelling was studied by superimposing protein density traces on a radioactive fractionation plot. These labelled proteins were incubated with spermatozoa and shown to bind more readily to spermatozoa from boars after the removal of the vesicular glands than to spermatozoa obtained from their normal litter mates. Most of the labelled protein became bound to the membranes which were isolated by sucrose density gradient centrifugation. The membranes were separated into two bands which equilibrated at the relative densities of 1·150 and 1·165. These fractions consisted of membrane vesicles of different size; the smaller band on the gradient, which equilibrated at 1·165, consisted of denser membrane material.

Because exposure to seminal plasma enhances the detrimental effects of cold shock on boar spermatozoa (Lasley & Bogart, 1944; Pursel, Johnson & Rampacek, 1972), many procedures for preserving semen by deep freezing employ sperm-rich ejaculate fractions (Pursel & Johnson, 1975; Larrson & Einarsson, 1975), thereby excluding much of the seminal plasma. Our previous studies showed that a basic protein fraction of seminal vesicle origin binds irreversibly to boar spermatozoa at ejaculation (Moore & Hibbitt, 1976) and during cooling promotes sperm membrane disruption (Moore, Hall & Hibbitt, 1976). We therefore tested the possibility that boar spermatozoa could be frozen more effectively in the absence of seminal vesicular fluid.

Summary.

Spermatozoa from intact boars and from boars without seminal vesicles were resuspended in diluent and cooled at different rates to 0°C. Glutamic oxaloacetic transaminase and lactate dehydrogenase activities were greater in the diluents which had contained spermatozoa from intact boars than in those which contained spermatozoa from animals without seminal vesicles. The incubation of seminal plasma from an intact boar with spermatozoa from a vesiculectomized animal before cooling also increased the enzyme activity in the diluent. The factors responsible for this effect were associated with the basic protein fractions of boar seminal plasma, in particular the proteins with haemagglutinating activity which may have been adsorbed onto the spermatozoa. Spermatozoa were exposed to colloidal Fe(OH)₂ ⁺ to determine by electron microscopy the charge on the surface of the plasma membrane of washed epididymal spermatozoa and ejaculated spermatozoa from intact and vesiculectomized boars. Epididymal spermatozoa bound the positively charged particles more readily than the ejaculated spermatozoa from the intact boars, due to the absence of membrane-bound protein.

Summary.

A technique is described for the removal of the seminal vesicles from the boar. The operation was carried out on twelve animals and six of the animals were subsequently trained for semen collection. The seminal plasma from the boars after surgery compared with normal litter mates had a more watery consistency and did not form the characteristic gel during ejaculation. The sperm concentration was 49% lower while the total reduction of sperm number/ejaculate was 78% in the experimental animals, but the ratio of living to dead spermatozoa remained unchanged. The concentrations of citrate and protein were significantly depressed in the seminal plasma of the animals after surgery and the pH increased; the osmolarity remained unchanged. Insemination of gilts with the semen from experimental boars revealed no significant loss of fertility compared with the normal controls. Animals slaughtered up to 17 months after surgery showed no regeneration of the seminal vesicles.

This study investigated the effects of short-term (20 days) ovariectomy, the effects of FSH assay (radioimmunoassay, receptor assay or in vitro bioassay) and of FecB^B genotype on the characteristics of pituitary FSH from Booroola ewes. Pituitary extracts were obtained from ovariectomized homozygous carriers (BB) and non-carriers (++, n = 8 per genotype) and ovary-intact controls (n = 4 per genotype). The extracts (n = 4 per genotype per treatment) were subjected to agarose suspension electrophoresis and the eluates were assayed by the three FSH methods. There were significant effects of ovariectomy (P < 0.01) and assay system (P < 0.05) but not of genotype on the median charge of FSH isoforms. The mean ± sem migration rates for FSH in intact and ovariectomized ewes were 0.469 ± 0.006 and 0.439 ± 0.006 albumin mobility units, respectively (P < 0.01), indicating a shift to more basic isoforms after short-term ovariectomy. When the pituitary extracts were subjected to anion-exchange HPLC, there was a significant (P < 0.01) shift to more basic isoforms in the ovariectomized ewes as shown using agarose electrophoresis, and no gene effects were noted. When the pituitary extracts (n = 4–8 per group) were injected into mature female mice, there were no significant effects of ovariectomy or genotype on the circulating half-lifes of the pituitary FSH isoforms. These results indicate that after short-term ovariectomy, the increase in plasma FSH concentrations is accompanied by a shift in the median charge of pituitary FSH isoforms without any observable change in their metabolic clearance rates. Moreover, the FecB^B gene has little effect on the median charge or half-life of pituitary FSH.

Summary. Peripheral blood samples were collected throughout pregnancy from 11 red deer hinds. During the same period, 6 other hinds which mated but failed to produce calves were also sampled. Pretreatment of some of these hinds included synchronization of oestrus alone (N = 3) or with injection of 1000 i.u. PMSG (N = 9).

During early and mid-pregnancy, LH and prolactin were frequently undetectable. Prolactin concentrations in pregnant and non-pregnant hinds were high (>250 ng/ml) in December—January. The results of the hormone analyses suggested that the amount of progesterone in plasma correlates with the number of corpora lutea (CL) present. The concentrations of oestradiol and progesterone were low from mid-winter onwards in the non-pregnant hinds, suggesting a reduction in ovarian activity at this time. In pregnant animals, progesterone concentrations were high for the first 200 days of gestation. Oestradiol rose to peak values concomitant with declining progesterone concentrations just before parturition.

Female spotted hyenas (Crocuta crocuta) have an erectile peniform clitoris and a pseudoscrotum but no external vagina, all established by day 35 of a 110-day gestation. Recent studies indicate that these events are androgen-independent, although androgen secretion by fetal ovaries and testis was hypothesized previously to induce phallic development in both sexes. We present the first data relating to the capacity of the ovaries and testes of the spotted hyena to synthesize androgens at different stages of fetal life. Specifically, spotted hyena fetal gonads were examined by immunohistochemistry at GD 30, 45, 48, 65, and 95 for androgen-synthesizing enzymes, as related to the morphological development. Enzymes included 17α-hydroxylase/17,20-lyase cytochrome P450 (P450c17), cytochrome b5, 3β-hydroxysteroid dehydrogenase (3βHSD), and cholesterol side-chain cleavage cytochrome P450 (P450scc). Anti-Müllerian-hormone (AMH) expression was also examined. AMH was strongly expressed in fetal Sertoli cells from GD 30 and after. P450c17 expression was detected in Leydig cells of developing testes and surprisingly in Müllerian duct epithelium. Fetal ovaries began to organize and differentiate by GD 45, and medullary cells expressed P450c17, cytochrome b5, 3βHSD, and P450scc. The findings support the hypothesis that external genital morphology is probably androgen-independent initially, but that fetal testicular androgens modify the secondary, male-specific phallic form and accessory organs. Fetal ovaries appear to develop substantial androgen-synthesizing capacity but not until phallic differentiation is complete, i.e. after GD 45 based on circulating androstenedione concentrations. During late gestation, fetal ovaries and testes synthesize androgens, possibly organizing the neural substrates of aggressive behaviors observed at birth in spotted hyenas. These data provide an endocrine rationale for sexual dimorphisms in phallic structure and reveal a potential source of androgenic support for neonatal aggression in female and male C. crocuta.