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Summary. Ovulatory effects of histamine and specific antagonists were studied in isolated perfused ovaries from immature rats treated with 10 i.u. PMSG to stimulate follicular growth and maturation. Histamine alone, like LH, induced ovulation in all ovaries tested, but the number of follicular ruptures was lower after histamine (7·0 and 2·2 ruptures, respectively, per ovary). The histamine-induced ovulations could be inhibited dose-dependently by the H₁-receptor antagonist, pyrilamine, or the H₂-antagonists, cimetidine and ranitidine. At the concentrations tested, these antagonists did not, when given separately, reduce the LH-induced ovulations significantly, but pyrilamine and cimetidine in combination lowered the ovulation frequency by 65%. The prostaglandin synthesis inhibitor, indomethacin, was not able to block the histamine-induced ovulations.

Summary. Mast cells, visualized with toluidine blue staining and the Falck-Hillarp fluorescence technique, were mainly located around large blood vessels in the hilus region of the ovary in adult rats and in immature rats treated with PMSG. Histamine concentration in the rat ovary was significantly reduced after the LH surge in PMSGtreated animals, corresponding to a reduced number of ovarian mast cells. No marked change in the number of mast cells and histamine concentration was found in adult rats during the oestrous cycle. Histamine as well as the H¹-agonist, 2-methylhistamine, and the H²-agonist, 4-methylhistamine, induced ovulations in the isolated perfused rat ovary. Ovulation rates were significantly lower than those evoked by LH. The histamine liberator, Compound 48/80, induced ovulations which were blocked by the combined effect of the H¹- and H²-histamine receptor antagonists, cimetidine and pyrilamine. The anti-degranulating agent, disodium cromoglycate, did not block ovulations induced by Compound 48/80.

The results show that the level of ovarian histamine, which is primarily stored in mast cells, can be influenced by PMSG treatment, and that the amine is able to induce ovulations in gonadotrophin-primed rats by an effect mediated by both H¹ and H² receptors.

Keywords: histamine; ovary; in vitro; ovulation

Sperm cells that cannot swim and orient properly compromise male fertility. Such defects are responsible for male infertility regardless of the actual quality of the most important content, the sperm’s DNA. Synthetic micromotors are engineered devices that are able to swim in (body) fluids and microscopic environments, similar to flagellated cells like sperm. Coupled together, a sperm-hybrid micromotor embodies the concept of bringing the sperm cell together with artificial components that assist or replace defective functions of the cell, helping it to pursue its goal without interfering with its health, enabling the process of assisted fertilization and further embryo development all inside the body. Non-invasive, remote-controlled in vivo applicability is the key quality of such hybrid microdevices. Assisted reproduction with the help of micromotors is in the focus of this review, although other biomedical applications that arise from the powerful combination of sperm cell and synthetic enhancement are also discussed and summarized. Details are provided about different fabrication processes and cell-material coupling strategies, and the way from proof-of-concept studies to in vivo experiments in animals is outlined.

Glycolysis is crucial for sperm functions (motility and fertilization), but how this pathway is regulated in spermatozoa is not clear. This prompted to study the location and the regulatory properties of 6-phosphofructokinase (PFK, EC 2.7.1.11), the most important element for control of glycolytic flux. Unlike some other glycolytic enzymes, PFK showed no tight binding to sperm structures. It could readily be extracted from ejaculated boar spermatozoa by sonication and was then chromatographically purified. At physiological pH, the enzyme was allosterically inhibited by near-physiological concentrations of its co-substrate ATP, which induced co-operativity, i.e. reduced the affinity for the substrate fructose 6-phosphate. Inhibition by ATP was reinforced by citrate and H⁺. Above pH 8, PFK lost all its regulatory properties and showed maximum activity. However, in the physiological pH range, PFK activity was very sensitive to small changes in effectors. At near-physiological substrate concentrations, PFK activity requires activators (de-inhibitors) of which the combination of AMP and fructose 2,6-bisphosphate (F2,6P₂) was most efficient as a result of synergistic effects. The kinetics of PFK suggest AMP, F2,6P₂, H⁺, and citrate as allosteric effectors controlling PFK activity in boar spermatozoa. Using immunogold labeling, PFK was localized in the mid-piece and principal piece of the flagellum as well as in the acrosomal area at the top of the head and in the cytoplasmic droplets released from the mid-piece after ejaculation.

Two experiments were conducted to evaluate the effect of follicular components on the maintenance of meiotic arrest in horse oocytes. In Expt 1, oocytes were incubated for 24 h with follicular fluid, or with granulosa cells suspended either in medium or in follicular fluid at 25 × 10⁶ cells ml⁻¹. None of the treatments resulted in significant maintenance of the germinal vesicle stage over that of non-suppressive control. Culture with follicular fluid plus granulosa cells resulted in a significantly higher proportion of oocytes at metaphase I compared with controls. In Expt 2, oocytes were divided into those originally having compact or expanded cumuli. Oocytes were cultured with sheets of mural granulosa or sections of follicle wall, or after injection into intact dissected follicles. After incubation, half of the oocytes from each suppressive treatment were matured for 24 h. All three suppressive treatments were effective in maintaining oocytes at the germinal vesicle stage (no significant difference from control oocytes fixed directly after removal from the follicle). However, no treatment maintained normal viability of oocytes, as significantly fewer oocytes were at metaphase II after all the suppression–maturation treatments compared with the maturation control. The highest rate of post-suppression maturation was found in the mural granulosa treatment. Within this treatment, the proportion of oocytes in metaphase II was significantly higher for oocytes with expanded than for oocytes with compact cumuli (31% versus 11%, respectively; P < 0.05). Suppression by injection into an intact follicle was associated with a lack of progression to metaphase II during subsequent maturation. These results indicate that follicular fluid alone does not suppress maturation of horse oocytes. Incubation of horse oocytes with follicle wall tissue (sheets of granulosa, follicle wall sections or intact follicles) can effectively suppress maturation but variably impairs the viability of oocytes.

Summary. The immature rat ovary contains VIP immunoreactive nerve fibres sparsely distributed around blood vessels, in the interstitial gland and around follicles. The VIP concentration, measured radioimmunologically, decreased significantly after PMSG treatment (10 i.u.), probably due to ovarian enlargement and oedema, while the total VIP content (total of 0·12 pmol in both ovaries) did not change after PMSG priming. The ovulatory effect of VIP was studied using in-vitro perfused ovaries from immature 28-day-old rats primed with 10i.u. PMSG. In all ovaries perfused, VIP (10⁻⁷ m) induced ovulations with a rate of 2·33 ± 0·56. The ovulation rate was significantly lower than that of ovaries stimulated by LH (0·1 μg/ml) (5·20 ± 0·86 ovulations per ovary). No synergistic effect on the ovulation rate was seen when LH and VIP were administered together (5·20 ± 0·49 ovulations per ovary). The results suggest that the neuropeptide VIP may represent one of the local factors involved in the ovulation process.

Keywords: VIP; rat ovary; ovulation