Immunohistochemical localization of taurine in the rat ovary, oviduct, and uterus.

The distribution of the amino acid taurine in the female reproductive organs has not been previously analyzed in detail. The aim of this study was to determine taurine localization in the rat ovary, oviduct, and uterus by immunohistochemical methods. Taurine was localized in the ovarian surface epithelium. The granulosa cells and oocytes of primordial follicles were immunonegative. In primary and antral follicles, taurine was found mainly in theca cells and oocytes, whereas the zona pellucida, antrum, and most granulosa cells were unstained. However, taurine immunoreactivity in theca cells and oocytes decreased during follicular atresia. During corpora lutea development, the number of immunopositive theca lutein cells increased as these cells invaded the granulosa-derived region. Therefore, most luteal cells from the mature corpora lutea were stained. In the regressing corpora lutea, however, taurine staining in luteal cells decreased. In the fimbriae, infundibulum, and uterotubal junction, taurine was localized in most epithelial cells. In the ampullar and isthmic segments, taurine was found in the cilia of most ciliated cells and in the apical cytoplasm of some non-ciliated cells. In the uterus, most epithelial cells were immunopositive during diestrus and metestrus, whereas most of them were immunonegative during estrus and proestrus. Moreover, taurine immunoreactivity in the oviduct and uterus decreased with pregnancy. (J Histochem Cytochem 49:1133-1142, 2001)     

Progesterone binding in rabbit oviduct and uterus.

Progesterone binding of high affinity with a dissociation constant of 10(-9) M was identified in cytosol of rabbit oviduct and uterus. Macromolecules with sedimentation coefficients of 7-8 S and 4-5 S were present. Progesterone receptor concentration was two to fivefold lower in the oviduct when compared with the uterus. The receptor concentration declined steadily from 3 hr until 144 hr after mating in the uterus; however, the decline in oviductal receptor was not significant until the sixth day of pregnancy. Progesterone receptor concentration in rabbit oviduct and uterus in estrus and early pregnancy was greater than estradiol receptor levels.     

Hormone-sensitive lipase expression and IHC localization in the rat ovary, oviduct, and uterus

Hormone-sensitive lipase (HSL) is a key regulator of cholesterol esters metabolism. The aim of this study was to determine HSL localization in rat female reproductive organs during the ovarian cycle by IHC methods. HSL was located in the ovarian epithelium. The granulosa cells and oocytes of primordial follicles were immunonegative. In mature follicles, HSL was found in oocytes and theca and granulosa cells. However, HSL expression in theca cells and oocytes decreased during follicular atresia. Luteal cells showed HSL staining in cytoplasm during proestrus and estrus, in the nucleus during metestrus, and in cytoplasm and the nucleus during diestrus. In the tubaric ampulla, HSL was located in the epithelial cells nuclei and in the cilia during proestrus and estrus but mainly in the nucleus during metestrus and diestrus. In the isthmus, cells showed HSL immunolabeling in the nucleus and cilia during proestrus, but only in the cilia during estrus, metestrus, and diestrus. In the uterus, HSL was found in the epithelial cells nuclei. HSL-immunoreactive bands at 84, 67, 54, and 43 kDa were found in rat female reproductive organs. HSL labeling in the nucleus of epithelial and germ cells suggests an as yet unknown function for this protein, probably related to oogenesis and cell proliferation.     

Ultrastructural and kinetic studies on uteroglobin secretion in the uterus and oviduct of the pseudopregnant rabbit

The intracellular localization of uteroglobin, a progesterone-induced protein, was studied in uterus and oviduct by means of immunoelectron microscopy with the protein A-gold technique. In the uterus, uteroglobin was synthesized in the columnar epithelium of the endometrium where most of the cells were immunoreactive. The protein was localized mainly in small secretory granules which were seen in the process of release into the uterine lumen. The luminal microvilli were also heavily stained. In the oviduct, the secretory cells contained large immunoreactive granules at the apical zone, some of which were observed while discharging into the lumen. Within these secretory granules, uteroglobin accumulated particularly in lens-shaped patches at the periphery of the granules. In vitro kinetic studies on the secretion of newly synthesized uteroglobin indicated that the ability to store uteroglobin is greater in the oviduct than in the uterus; however, the rate of uteroglobin secretion is greater in the uterus than in the oviduct. Thus, there appears to be a good correlation between the microscopic and the functional observations.     

Principal features of the nerve supply to the ovary, oviduct and tubal third of the uterus in the golden hamster

The autonomic outflow and sensory structures in the ovary and accessory reproductive organs of the hamster are described by means of specific fluorescence and enzyme histochemical techniques for the demonstration of catecholamine and acetylcholinesterase (AChE), respectively. Sympathetic nerves accompany branches of the major blood vessels in the mesentery of the ovary, oviduct and tubal uterine horn and invest the vascular bed in each of these organs. Vasomotor fibers predominate in the ovary and oviduct, though occasional adrenergic axons supply thecal and interstitial tissues in the ovary and the longitudinal smooth muscle of the oviduct. Fluorescent myomotor axons run in the suspensory ligament and outer myometrial layer of the uterus, but most of the numerous sympathetic and AChE-fibers in the tubal third of the horn supply the intramural and submucosal vascular plexuses. A limited electron microscopic study of the central spiral (preplacental) arteries of the endometrium indicates that the surrounding terminal AChE-fibers are identical to the fluorescent and granular vesicle-bearing adrenergic axons which form neuromuscular junctions with these vessels. Based on the discovery of specialized sensory endings in the walls of the large collecting veins which drain the hamster uterus, a mechanism is proposed to account for the regulation of blood flow through maternal placental vessels which are devoid of an arteriolar neuromuscular apparatus.     

Existence and coexistence of peptides in nerves of the mammalian ovary and oviduct demonstrated by immunocytochemistry

Summary The immunocytochemical distribution of substance P (SP), gastrin releasing peptide (GRP), vasoactive intestinal polypeptide (VIP), peptide histidine isoleucine (PHI), and neuropeptide Y (NPY) was studied in the ovary and the Fallopian tube (oviduct) of rats, guinea-pigs, cows, pigs and humans. Generally, the nerve supply was better developed in the oviduct than in the ovary. GRP fibers were most scarce in all tissues. Nerves containing SP were particularly numerous in the oviduct of rat and guinea-pig, supplying the muscular wall and blood vessels. VIP and PHI coexisted in dense plexuses of nerves, not only around blood vessels but also in the follicular wall and the interstitial gland of the ovary, as well as within the smooth muscle layers and subepithelially in the oviduct. The general distribution of NPY was similar, but these immunoreactive nerves were even more numerous. Sequential staining for dopamine--hydroxylase and NPY together with results of chemical sympathectomy with 6-hydroxydopamine suggested that NPY was stored in the noradrenergic sympathetic nerves.     

Existence and coexistence of peptides in nerves of the mammalian ovary and oviduct demonstrated by immunocytochemistry

The immunocytochemical distribution of substance P (SP), gastrin releasing peptide (GRP), vasoactive intestinal polypeptide (VIP), peptide histidine isoleucine (PHI), and neuropeptide Y (NPY) was studied in the ovary and the Fallopian tube (oviduct) of rats, guinea-pigs, cows, pigs and humans. Generally, the nerve supply was better developed in the oviduct than in the ovary. GRP fibers were most scarce in all tissues. Nerves containing SP were particularly numerous in the oviduct of rat and guinea-pig, supplying the muscular wall and blood vessels. VIP and PHI coexisted in dense plexuses of nerves, not only around blood vessels but also in the follicular wall and the interstitial gland of the ovary, as well as within the smooth muscle layers and subepithelially in the oviduct. The general distribution of NPY was similar, but these immunoreactive nerves were even more numerous. Sequential staining for dopamine-beta-hydroxylase and NPY together with results of chemical sympathectomy with 6-hydroxydopamine suggested that NPY was stored in the noradrenergic sympathetic nerves.     

Uptake of labeled mammalian gonadotropins by ovary and oviduct of the lizard, Lacerta s. sicula, in vivo

The uptake of radioactivity by ovary, oviduct and thigh muscles of the lizard, Lacerta s. sicula, after administration of labeled mammalian gonadotropins has been followed. Ovary and oviduct show a significantly higher radioactivity than thigh muscles. The ovarian uptake, moreover, is decreased by the corresponding non-labeled gonadotropin. The meaning of these observations for the physiological regulation of reproductive processes in that lizard, are discussed.     

Relationship between the electrical activity of the oviduct and the uterus of the rabbit in vivo.

The electrical activity of the whole genital tract of the rabbit was recorded by means of chronically implanted electrodes after section of the uterotubal junction on one side. When the junction was intact, the activity of the isthmus and that of the proximal uterine horn occurred almost simultaneously, but uterine activity decreased after the junction was cut. During the preovulatory phase and also after administration of HCG, synchronos activity due to adrenergic drugs, smoke or oxytocin persisted on both sides of the uterotubal junction. Hypersensitivity of the isthmus and the proximal segment of the uterine horn was recorded on the cut side after ovariectomy. The concept of a local control mechanism in the region of the uterotubal junction with a positive control of the uterus by the oviduct is suggested.     

Regional differences in expression of progesterone receptor in oviduct and uterus of rabbit during early pregnancy

We characterized the expression pattern of progesterone receptor (PR) in two regions of the oviduct (ampullae and isthmus), and the uterus (epithelium and stroma) of the rabbit (Oryctolagus cuniculus) during early pregnancy (1-4 days) by RT-PCR and immunohistochemistry. We observed a significant increase in the expression of PR at mRNA level in the uterus on days 1 and 2 of pregnancy, followed by a decrease on days 3 and 4. These changes were also observed at protein level in the uterine epithelium. Interestingly, PR immunoreactivity decreased in stromal cells in all days of pregnancy as compared with non-pregnant rabbits (NG). In the isthmus PR mRNA expression significantly increased on day 2 of pregnancy and diminished on days 3 and 4, whereas no significant changes were observed in the ampullae. In epithelial and stromal cells of the isthmus, PR immunostaining was reduced through pregnancy as compared with NG group. In contrast, a reduction in PR immunostaining was observed on days 1-3 with an increase on day 4 in epithelial and stromal cells of the ampullae. The overall results suggest that PR exhibit a differential expression pattern in the oviduct and the uterus during early pregnancy of the rabbit, and that these differences are related to different functions of PR in the reproductive tract during early pregnancy.     

Co-culture of rabbit 2-cell embryos with rabbit oviduct epithelial cells and other somatic cells

Rabbit 2-cell embryos were co-cultured in Basel Synthetic Medium II + 10% fetal bovine serum with one of the following: primary cultures of rabbit oviduct epithelial cells (ROEC), a rabbit kidney epithelioid cell line (RK13), a rabbit epidermal epithelioid cell line (Sf1), or a rabbit skin fibroblast-like cell line (RAB9). Embryos cultured in medium alone served as controls. After 4 d of culture at 39 degrees C in 5% CO2 in air, 77-93% of the rabbit embryos which were co-cultured with somatic cells had reached the blastocyst stage, and 60-76% were hatching through their zonae pellucidae. These percentages, however, were not significantly different (P greater than .05) from those of embryos in medium alone, of which 90% had reached the blastocyst stage and 83% were hatching. Mean intrazonal embryo diameters also did not differ significantly among treatments (239-302 microns). Bovine 1-8-cell embryos were also co-cultured with ROEC. This stimulated 60% of these embryos to develop beyond the so-called "16-cell block" in vitro, whereas 0% of the embryos cultured in medium alone developed past this block. Evaluation of the ROEC cultures by light microscopy, immunocytochemistry, and gel electrophoretic analysis of conditioned medium, together with the positive results with bovine embryos, indicate that the ROEC culture partially simulates oviductal conditions in vivo. Therefore, our results suggest that oviduct epithelial cells may play a less pivotal role in regulating early development in the rabbit than in the cow.(ABSTRACT TRUNCATED AT 250 WORDS)     

Relationship between cellular DNA synthesis,PCNA expression and sex steroid hormone receptor status in the developing mouse ovary,uterus and oviduct

The proliferative activities of the different cellular compartments of the developing mouse ovary, uterus, and oviduct were studied by radioautographic assessment of DNA synthesis with [³H]-thymidine labeling and by immunohistochemical staining of proliferating cell nuclear antigen (PCNA). The distributions of estrogen and progesterone receptors (ER and PR) were studied by immunohistochemical staining. The values of the PCNA positive staining indices were a little higher than that of the radioautographic labeling indices. However, linear relations were shown for the two indices. The proliferative activities were high from postnatal day 1–7 and decreased from day 14 in the different cellular compartments of the ovary. The proliferative activities were high on days 1, 3 and decreased from day 7 in the uterus and oviduct. Staining of ER and PR was very weak in the surface epithelium, stroma and large follicles of the ovary. Positive staining for ER occurred from day 14 in the uterine epithelium and from day 7 in oviductal epithelium. Positive staining for PR was observed from day 1 in both the uterine and oviductal epithelium. However, the positivity of both ER and PR occurred from postnatal day 1 in the stromal cells of the uterus and oviduct. These results suggest that the appearance of the steroid receptors differ between the different cellular compartment of the reproductive organs. The proliferative activities have an inverse relation with the expression of the steroid hormone receptors in the female reproductive organs during developmental stages. Therefore, we propose that there is an autonomous proliferation mechanism in the development of the reproductive organs or that the proliferation is moderated by factors other than steroid hormones.     

New gene assignments in the rabbit (Oryctolagus cuniculus). Comparison with other species

Nineteen cell hybrids were obtained by fusing rabbit (Oryctolagus cuniculus, OCU) fibroblasts and a Chinese hamster cell line HGPRT-. Eleven enzymatic markers were previously investigated (Soulié and Grouchy 1982); seven of these could be assigned (LDHA, LDHB, TPI, PEPB, NP, ITP, and G6PD). Two assignments were uncertain (MDH2 and GUK). Two markers could not be assigned (MDH1 and PGD). Seven further markers were investigated and are the subject of this report. Six could be assigned: GALT to chromosome OCU1, GAPD to OCU4, GPX and ACY to OCU9, PGM1 to OCU13, and GSR to OCU19. One could not be assigned (GPI). MDH2 and GUK were previously considered uncertain. Now MDH2 was found impossible to assign and GUK was mapped on OCU15. These assignments were compared with those known in man, Cebus capucinus, Microcebus murinus, cat, and mouse. It was impossible to assign any enzymatic marker belonging to the ten linkage groups known in the rabbit. The esterase locus could not be investigated since the rabbit enzyme migrates in the same position as the hamster enzyme.     

Influence of hCG injection and steroid treatment on prostaglandin metabolism by rabbit uterus and oviduct

Metabolism of PGE-2 and PGF-2 alpha by cytosolic fractions (100 000 g supernatant) of rabbit uterus, oviduct and lung was measured in vitro. Metabolism of PGE-2 was greater than that of PGF-2 alpha for oviduct and uterus. After an ovulating injection of hCG metabolism of both PGE-2 and PGF-2 alpha by lung and uterus declined linearly up to 72 h (during the time of ovum transport). The amount of PG metabolism by the oviduct did not change significantly during this period, but the percentage changes of PGE-2 and PGF-2 alpha metabolism from oestrous values did differ, and perhaps indicated a change in the ratio of intracellular PGs. No change of metabolism of either PG by lung, uterus or oviduct occurred at 24 or 72 h after an injection of 250 micrograms oestradiol cyclopentylpropionate given concomitantly with the hCG (a treatment regimen which causes 'tube-locking' of ova). However, progesterone treatment, in a regimen known to cause accelerated transport of ova through the oviduct, caused significantly enhanced metabolism of both PGE-2 and PGF-2 alpha by uterus and oviduct, but not lung, 30 and 48 h later except for PGE-2 by uterus at 30 h. These results suggest that changes in metabolism of PGE-2 and PGF-2 alpha by the oviduct may be involved in the mechanisms controlling ovum transport.     

Autotransfer of Day 4 embryos from oviduct to oviduct versus oviduct to uterus in the mare

Embryo autotransfer is defined as the collection of an embryo from and the transfer of this embryo into the same animal. The objectives of this study were to: 1) test the hypothesis that oviduct transport of the equine embryo from the oviduct into the uterus is not dependent on a unilateral embryo-corpus luteum interaction, 2) develop an embryo autotransfer technique for the mare and 3) compare the success rates of Day 4 embryos surgically autotransferred from the oviduct ipsilateral to ovulation to either the oviduct (n=10 mares) or the uterine horn (n=10 mares) contralateral to ovulation. Seventy percent (7 10 ) of the Day 4 embryos which were autotransferred to the oviduct contralateral to ovulation were transported through the oviduct and subsequently developed into embryonic vesicles detectable by ultrasonography between 10 and 21 days postovulation. This finding supported the hypothesis that oviductal embryo transport is not dependent upon the ipsilateral corpus luteum. Overall, sixty percent (12 20 ) of the autotransfers were successful. The success rate of uterine-transferred embryos was not significantly less (P>0.3) than that of oviductal-transferred embryos (5 10 vs 7 10 , respectively). Therefore, the Day 4 equine embryos were apparently mature enough to survive in the mare's uterus.