Cloprostenol induced luteal regression in the beef cow. II. Histological evaluation of corpora lutea and correlation with luteal progesterone

Forty-two cycling, multiparous beef cows (percentage-Brahman) were injected twice at 11-d intervals with 500mug Cloprostenol (a prostaglandin F(2alpha) analog) to induce luteolysis. Cows were randomly assigned for ovariectomy at 12 hr intervals from 0 to 72 hr post-injection. Corpora lutea were excised and one-half of the corpus luteum was stored in phosphate-buffered formalin until mounting and staining with hematoxylin and eosin. The other half of the CL was snap-frozen for determination of progesterone content and concentration. Luteal cell density increased following Cloprostenol injection and was significantly correlated with a shift from predominantly healthy Type I cells to predominantly degenerating Types III and V cells. Cell mitosis tended to decrease by 12 hr and was lower by 24 hr post-injection. Cell pyknosis increased by 24 hr post-injection and was correlated with the decrease in percentage of healthy luteal cells. No pattern was detectable in cell karyorrhexis. Histological regression of the CL was inversely correlated with CL progesterone content. Therefore, we conclude that a reduction in cell mitosis is the earliest morphological sign of degeneration of the CL and that the CL follows a well-defined sequence of regression which is accompanied by a decrease in progesterone content.     

Variation in the oxytocin content of caprine corpora lutea across the breeding season

Ovarian tissues were obtained from cyclic goats during the early, mid and late breeding season. Immunoreactive oxytocin was measured by RIA in tissue extracts after chromatography on octadecylsilica cartridges. Luteal oxytocin concentrations were significantly greater during the early breeding season than during the mid or late breeding season. Oxytocin is luteolytic in goats. High concentrations of luteal oxytocin may be related to the high incidence of short estrus at the onset of the breeding season.     

Gap junctional intercellular communication of bovine luteal cells from several stages of the estrous cycle: Effects of prostaglandin F2α, protein kinase C and calcium

Cellular interactions mediated by both contact-dependent and contact-independent mechanisms are probably important to maintain luteal function. The present studies were performed to evaluate the effects of luteotropic and luteolytic hormones, and also intracellular regulators, on contact-dependent gap junctional intercellular communication (GJIC) of bovine luteal cells from several stages of luteal development. Bovine corpora lutea (CL) from the early, mid and late luteal phases of the estrous cycle were dispersed with collagenase and incubated with no treatment, LH, PGF or LH + PGF (Experiment 1), or with no treatment, or agonists or antagonists of protein kinase C (TPA or H-7) or calcium (A23187 or EGTA; Experiment 2). After incubation, media were collected for determination of progesterone concentrations. Then the rate of GJIC was evaluated for small luteal cells in contact with small luteal cells, and large luteal cells in contact with small luteal cells by using the fluorescence recovery after photobleaching technique and laser cytometry. Luteal cells from each stage of the estrous cycle exhibited GJIC, but the rate of GJIC was least (P<0.05) for luteal cells from the late luteal phase. LH increased (P<0.05) GJIC between small luteal cells from the mid and late but not the early luteal phase. PGF increased (P<0.05) GjIC between small luteal cells from the mid luteal phase and diminished (P<0.05) LH-stimulatory effects on GjIC between small luteal cells from the late luteal phase. Throughout the estrous cycle, TPA decreased (P<0.05) the rate of GjIC between large and small, and between small luteal cells, and A23187 decreased (P<0.05) the rate of GJIC between large and small luteal cells. LH and LH + PGF, but not PGF alone increased (P<0.05) progesterone secretion by luteal cells from the mid and late luteal phases. Agonists or antagonists of PKC or calcium did not affect progesterone secretion by luteal cells. These data demonstrate that both luteal cell types communicate with small luteal cells, and the rate of communication depends on the stage of luteal development. LH and PGF affect GjIC between small luteal cells during the fully differentiated (mid-luteal) and regressing (late luteal) stages of the estrous cycle. In contrast, at all stages of luteal development, activation of PKC decreases GjIC between small and between large and small luteal cells, whereas calcium ionophore decreases GjIC only between large and small luteal cells. Luteotropic and luteolytic hormones, and intracellular regulators, may be involved in regulation of cellular interactions within bovine CL which likely is an important mechanism for coordination of luteal function.     

Continuous culture of Humicola lutea 120-5 for acid proteinase production

Acid proteinase production by the fungus Humicola lutea 120-5 in continuous culture was studied. The maximum activity of the culture broth reached 2200 U/ml at a dilution rate (D) of 0.05/h. The continuous process was carried out for 1 month without any bacterial contamination, due to low pH (3.0–3.5) during the cultivation.     

Short-lived corpora lutea syndrome in anoestrous ewes following 17β-oestradiol or MAP treatments applied before an allogenic sexual stimulation with rams and oestrous ewes

When induced to ovulate during anoestrus, ewes, does and cows frequently develop a short-lived corpora lutea (SLCL) syndrome associated to lack of previous progesterone. Exogenous progesterone precludes SLCL by blocking oxytocin endometrial receptors, thus inducing normal life-span CL (NLCL). Paradoxically, circa 50% of unprimed ewes do not develop SLCL. We report results from 3 trials assessing follicular, oestrous, ovulatory, and luteal end-points after 17β-oestradiol or MAP treatments. Oestradiol benzoate (50 μg) induced follicular turnover, provoked ovulation in 40% (24/60) of ewes treated (93% of which developed SLCL), but did not affect the incidence of SLCL (26/53) after an allogenic sexual stimulation (ASS) by rams and oestrous ewes. By the onset of the ASS, most NLCL ewes (26/27) had already experienced turnover of their largest follicle, had smaller largest and second largest follicles, and ovulated their largest follicle more frequently than SLCL ewes did. Most SLCL ewes (19/25) did not ovulate their largest follicles, ovulating instead smaller follicles of identical size to those of NLCL ewes. Priming (40 mg of MAP for 12 days) was partially effective at preventing SLCL even when terminated 14 days in advance of an ASS, but failed at completely preventing SLCL when terminated 6 or more days in advance. The coupling of a timed acquisition of full steroidogenic capability before ovulation with a system of endometrial oestradiol–progesterone–oxytocin receptors linked in an unstable equilibrium controlling the amplification of the luteolytic feed-forward loop of oxytocin and prostaglandin F₂α explains occurrence and relative incidences of both NLCL and SLCL, and links proximate and ultimate causes of the SLCL syndrome.     

Histology,histochemistry and morphometry of the ovary of the adult plains viscacha (Lagostomus maximus) in different reproductive stages

Lagostomus maximus is a South American Hystricognathi rodent whose reproductive behaviour shows characteristics unusual for mammals, such as polyovulation (200–800 oocytes) and a high rate of embryo mortality. Thirty‐six mature females captured in the province of Buenos Aires showed different physiological reproductive stages. Most of them presented a postpartum oestrus in August–September. This characteristic is different from that reported in other geographical areas. The stages considered were: anoestrus, follicular phase, early pregnancy and late pregnancy. The ovaries were light‐pink and smooth and presented a tortuous cord‐like aspect. Many primordial follicles were found in all the females studied. Follicles in different states of maturation and primary and accessory corpora lutea were observed in the cortex. These structures were smaller than those present in other related species. Follicles did not project into the surface of the organ. Calcified follicles of diverse size were found in all the ovaries. Atretic follicles were found in all the stages analysed. Interstitial tissue surrounding the follicles and the corpora lutea was also observed. The number and proportion of different cortical structures varied in the physiological stages analysed. The ovaries of the viscacha have differential characteristics in comparison to other Hystricognathi, some of them related to polyovulation.     

Performance of Trichogrammatoidea sp. nr lutea Girault (Hymenoptera: Trichogrammatidae) collected from Mussidia spp. in Kenya on eggs of six lepidopteran hosts

Mussidia nigrivenella Ragonot (Lepidoptera: Pyralidae), an important pest of maize ears in West Africa, has never been reported to attack crops in East and southern Africa (ESA), though it was found on various wild host plants in these regions. It was suggested that in ESA M. nigrivenella might be under natural control. In Kenya, exploration for natural enemies associated with Mussidia spp. yielded several parasitoids including a trichogrammatid egg parasitoid, Trichogrammatoidea sp. nr lutea Girault. The ability of T. sp. nr lutea to attack the eggs of several lepidopteran species found in Kenya was studied. The lepidopterans included the noctuids Busseola fusca (Fuller) and Sesamia calamistis (Hampson), the pyralids Eldana saccharina Walker, Mussidia fiorii Cecconi and de Joannis and Mussidia‘madagascariensis’, and the crambid Chilo partellus (Swinhoe). The former three species also infest cereals in West Africa. Trichogrammatoidea sp. nr lutea successfully attacked and developed in eggs of all six species indicating its potential to exploit other lepidopteran pests of maize in West Africa. Busseola fusca and S. calamistis were the most suitable hosts and had the largest number of eggs parasitized and progeny per female wasp where E. saccharina and C. partellus were the poorest hosts. The host species used to rear the parasitoid and the age of egg also significantly affected the total number of host eggs parasitized by the parasitoid. It was concluded that the ability of T. sp. nr lutea to exploit lepidopterans that are also pests of maize in West Africa may enhance biological control of M. nigrivenella and it should be considered for translocation to that area from Kenya.     

The pattern of ovulation in the southern African spiny mouse (Acomys spinosissimus)

The pattern of ovulation in mammals is generally considered to be either spontaneous or induced by coitus. The present study aimed to assess the pattern of ovulation in the southern African spiny mice (Acomys spinosissimus). Females were divided into three treatments differing in the degree of contact with a male. Control females had no contact with males, separated females had only chemical, auditory and visual contact with a male as the sexes were separated with wire mesh and paired females had full contact with a vasectomized male and copulations were possible. Each treatment consisted of seven females and the ovarian mass, the number of primary, secondary/tertiary and Graafian follicles as well as presence of corpora lutea were compared between the three treatments. Faecal progestagen metabolite (FPM) concentrations were analyzed for every second day throughout the experiment and they were used to determine luteal phases and oestrous cycles. Corpora lutea were found in both the control and the paired treatment indicating that ovulation also occurred in the absence of coitus. There was also no effect of treatment on ovarian mass or follicle numbers. In contrast, only females in the separated and paired treatments exhibited luteal phases and oestrous cycles. Especially at the beginning of the experiment, FPM concentrations were higher in those two groups than the control. The results indicate that A. spinosissimus appears to ovulate spontaneously, although physical as well as olfactory male cues appear to be of great importance to enhance reproductive efforts of females.