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In spite of the importance of the corpus luteum in human reproduction, little is known about its formation after ovulation and during regression in the absence of conception. This is largely due to constraints on the availability of normal human tissue; therefore an appropriate model which could be studied and could provide information applicable to the human was sought. The baboon (Papio), a non-human primate, has been determined to be one such model. Thus, in the past several years our studies have examined the role of luteal peptides in corpus luteum function, and, when possible, we have attempted to examine corpora lutea from the human and baboon in parallel. Although a milk-ejection factor was recognized to be present in luteal tissue in 1910 (Ott and Scott, Proc. Soc. Exp. Biol. Med., Vol. 8, p. 49), the role of oxytocin in luteal physiology has not been easy to ascertain. This is in part due to the methodologies employed to assess its role. Our studies summarized below suggest that oxytocin does not directly affect luteal steroidogenesis but that it may play a role in cell to cell communication involving the expression of the gap junction proteins, the connexins. In view of the fact that oxytocin, its receptor, gap junctions and associated proteins are not unique to the human and non-human primates, the model of luteal development and demise proposed may be applicable to most species.

Objective: To present our current understanding of oxytocin and its receptors during pregnancy and parturition and their potential clinical applications., Data Sources: A MEDLINE search was conducted for pertinent articles from 1966 to October 1996 related to oxytocin and its receptor and their clinical implications during pregnancy and parturition. Review articles, book chapters, and published trials were also searched., Methods of Study Selection: Only references in English that were deemed relevant were used. When possible, human data and sometimes animal data pertinent to understanding the interaction of oxytocin and its receptors were selected., Tabulation, Integration, and Results: Oxytocin is synthesized in the hypothalamus and in many reproductive tissues during pregnancy, whereas the receptors are synthesized in reproductive tissues. The genes for oxytocin and its receptors are on chromosomes 20 and 3, respectively. Oxytocin and its receptors are regulated by sex steroids and by oxytocin itself. The paracrine and autocrine mechanisms regulating oxytocin and its receptor within the fetoplacental-uterine unit are central to the control of uterine contractions and parturition. Such current understanding provides the basis for appropriate oxytocin regimens to induce or augment labor, to inhibit preterm labor by blockade of oxytocin receptors, and to achieve cervical ripening., Conclusion: Advances in our knowledge of oxytocin and its receptor have provided rational and sound principles for current concepts about their role in parturition, the appropriate use of oxytocin to stimulate the pregnant uterus or ripen the cervix, and the use of oxytocin antagonist to inhibit uterine contractions and preterm labor.

Although oxytocin has been recognized as a product of the corpus luteum in numerous species, including nonhuman primate and women, for sometime, its precise role in luteal physiology has remained obscure. However, with the recent observations that the steroidogenic activity of the large and small cells is increased in the presence of LH when these cells are in intimate contact has led to the hypothesis that cell-to-cell communication must occur between these cells. Cell-to-cell communication is possible via several mechanisms, including paracrine/autocrine and intercellular crosstalk via gap junctions. Substantial morphological and immunohistological evidence using antibodies to gap-junction specific proteins, the connexins, indicates the presence of gap junctions in corpora lutea. Our recent studies indicate that oxytocin affects the expression of the gap-junction protein connexin-43 and that the gonadotropins are intimately involved in this action. The synthesis of oxytocin and the oxytocin receptor is influenced by the gonadotropins and locally produced prostaglandins. Oxytocin stimulates estradiol synthesis, which may affect the expression of the gap-junction protein connexin-43, allowing interaction between the large cells and small cells of the corpus luteum. With the ubiquitous presence of oxytocin and its receptor, and the presence of gap junctions in the corpora lutea of numerous species, it is concluded that oxytocin is involved in not only paracrine/autocrine interaction but also may be of significant importance in intercellular crosstalk in the corpus luteum.

Baboon corpora lutea (two each from the early, mid- and late luteal phases) were individually microretrodialyzed in vitro for 48 h, 12 h initial baseline, 12 h retrodialysis with OT (9 mU/h), 12 h without OT and 12 h with cAMP (5 mmol/h). Progesterone (P) was measured by a sensitive and specific radioimmunoassay in 10-min fractions of retrodialysates and analyzed for P peaks by PC-pulsar 3.0. Neither OT nor cAMP had any effect on the characteristics of P pulses. In early and late luteal phase CL, OT inhibited P secretion within 1 h of administration followed by increased P secretion late during OT perfusion. In midluteal phase, OT did not affect P secretion. In all CL, P secretion was sustained or further increased during the 12 h after stopping OT. cAMP also sustained baseline or stimulated P secretion. In contrast, OT either increased total P output/12 h (28 to 49% above baseline) with a further increase of 21% to 296% above baseline after stopping OT, or inhibited total P output by 4% to 13% percent with a further decline of 51% to 61% after stopping OT. Thus, while overall OT is luteotropic, its dual effect (initial inhibition followed by stimulation) suggests direct and indirect effects through paracrine-autocrine mechanisms.

Oxytocin has been identified in both non-human primate and human corpora lutea of the menstrual cycle by RIA, immunocytochemistry and HPLC. Evidence for the transcription of the oxytocin gene in this tissue using PCR is available. Oxytocin receptors have been characterized by biochemical procedures. However, there is some debate as to whether the oxytocin identified in these tissues is biologically active and has a role in luteal function. In this study we have demonstrated that oxytocin isolated by gel chromatography of tissue extracts from the baboon and the human corpus luteum is biologically active as determined in a rat uterine bioassay. Since both oxytocin and its receptors are present in these tissues, it is suggested that oxytocin in the human and non-human primate corpora lutea has a functional role.

Although oxytocin (OT) was identified in the human and primate corpus luteum (CL) over a decade ago, a definitive role for this peptide has not been elucidated. Several in vitro models have been utilized to examine the most obvious role for OT in luteal function, that of its effect on progesterone (P) production. Using dispersed cells in short term incubations, cultured cells and microdialysis procedures utilizing intact tissue, variable effects of OT on P production have been obtained. We therefore hypothesized that OT may have other role(s) in this tissue. The follicle cells remaining after a successful ovulation has occurred undergo rapid luteinization and the tissue is extensively remodeled to form the CL. At this time, in most species, two types of luteal cells have been identified based on their morphology, biochemistry and size. Both cell types produce P, however only one cell type responds to the long distance modulator LH. Since both cell types appear to be needed for the overall synthesis of P in adequate quantities required for the preparation of the endometrium "information trafficking" must occur between the two cell types. Our recent studies have shown the presence of gap junctions and connexin-43, E-cadherin, an adhesion molecule and ZO-1 protein associated with tight junctions in the baboon and human CL. The effect of OT on connexin-43 in the baboon CL has been examined.

A variety of molecular techniques were used to search human and baboon gonadal tissues for evidence of transcription of the genes for the peptide hormones oxytocin and vasopressin. Only a highly sensitive assay based on a modification of the polymerase chain reaction succeeded in detecting mRNA copies of the oxytocin gene in both human and baboon corpus luteum. Vasopressin gene transcription was not detected in human testis and corpus luteum and was found only once in four different experiments in baboon corpus luteum. Evidence for oxytocin gene transcription in the human testis was found in three of five experiments. The method employed and subsequent sequence analysis of the polymerase products verified the presence of oxytocin mRNA with normal hypothalamic-type exonic structure in primate corpus luteum. Nevertheless, the very low levels of mRNA present are unlikely to support other than local functions for the encoded nonapeptide hormones.

Corpora lutea, corpora albicantia, and ovarian stroma from normal human premenopausal ovaries were examined for the presence of oxytocin and neurophysin by using highly specific antisera and peroxidase-antiperoxidase light-microscopic immunohistochemistry. Oxytocin and neurophysin immunoreactivity was found in some but not all cells of the corpora lutea obtained on days 19 to 24 of the menstrual cycle. Stromal tissue and corpora albicantia did not give a positive reaction for either of these peptides, and negative results were also obtained with corpora lutea of mid- and term-pregnancy and preovulatory follicles. Specificity of the immunohistochemical reaction was confirmed by immunoabsorption tests. The specific localization of immunoreactive oxytocin and neurophysin in corpora lutea of the human menstrual cycle directly demonstrates the presence of oxytocin- and neurophysin-positive cells within the human corpus luteum.

Seventeen human fetal and neonatal pituitary glands were removed at necropsy, and analyzed for oxytocin content by a specific and sensitive radioimmunoassay, after homogenization in 0.4M acetic acid. Serial dilution of the pituitary extract showed parallelism with the oxytocin standard curve on the radioimmunoassay. Column chromatography of the pituitary gland extract gave a single peak of immunoreactive oxytocin as determined by the radioimmunoassay. In eight fetuses, 14 to 17 weeks' gestation, pituitary gland oxytocin was 10.2 +/- 5.9 ng/gland (mean +/- SE), whereas an 18-week fetus had 5.8 ng of oxytocin per gland. Pituitary gland oxytocin content increased to 38.4 ng/gland in a 20-week fetus removed at hysterectomy, and 31.6 ng/gland in a 26-week fetus. Fetal pituitary gland oxytocin values were 22.1 ng/gland and 57.0 ng/gland at 32 weeks and increased significantly to 544.3 +/- 33.8 ng/gland in 1- to 5-day-old term newborn infants (n = 3). However, a 13-day-old term newborn infant had 3.7 ng of oxytocin per pituitary gland. The increased pituitary gland oxytocin content with advancing gestation was due to a significant increase in oxytocin concentration rather than to an increase in the weight of the pituitary gland. The findings indicate that oxytocin is present in fetal pituitary glands as early as 14 to 17 weeks' gestation and increases at term to 50 and 13 to 14 times more than in early midtrimester and early third-trimester pregnancy, respectively.

Ovarian tissues (n = 26) obtained at surgery were assayed for oxytocin (OT) concentrations in different parts of the ovary by a specific and sensitive RIA after homogenization and extraction with 0.4 M acetic acid. Chromatography of the extract on a Sephadex G-25 column revealed a single peak identical to synthetic OT, as measured by RIA. Corpora lutea of the menstrual cycle had 10.8-53.0 ng immunoreactive OT/g tissue (n = 7), while those of early pregnancy had a concentration of 106.0 ng/g (n = 1). Ovarian stromal tissue had either undetectable or lower concentrations of OT (0-21.0 ng/g; n = 5) than the corpus luteum from the same ovary. While a luteoma of term pregnancy (n = 1), a benign cystadenoma (n = 2), and an endometriotic cyst (n = 1) had no detectable immunoreactive OT, the concentrations of immunoreactive OT were 20.0 ng/g in a thecoma, 1.4, 20.0, and 60.0 ng/g in preovulatory follicles (n = 3), and 41.0 and 37.0 ng/g in polycystic ovaries (n = 2). In one patient with premature ovarian failure, the ovaries had 9.0 ng/g and undetectable immunoreactive OT. These findings indicate the presence of immunoreactive OT in human ovaries, with significant concentrations in the corpus luteum and preovulatory follicles. It is probable that these tissues produce OTs or an OT-like material which may function as an ovarian luteolytic agent.

Oxytocin concentrations were determined in baboon (Papio anubis) corpora lutea, and the effect of oxytocin on dispersed luteal cell progesterone production was evaluated. Oxytocin concentrations increased significantly from an early luteal phase value of 2.1 +/- 1.1 ng/gm to a peak concentration of 18.1 +/- 4.3 ng/gm wet weight in midluteal phase corpora lutea. Corpora albicantia and ovarian stroma had comparatively low oxytocin concentrations. Reverse-phase high-pressure liquid chromatography of corpora lutea extracts gave a peptide peak (retention time, 17.25 min) similar to a standard oxytocin peak. Plasma oxytocin levels, which were significantly higher in the ovarian vein draining a corpus luteum than in the contralateral side or the femoral vein, declined significantly after luteectomy. Oxytocin was localized by immunocytochemical methods in luteal cells. In the early luteal phase oxytocin (4 to 800 mU; 1 mU is equivalent to 2 ng) inhibited basal and human chorionic gonadotropin-stimulated progesterone production by dispersed luteal cells, but in the late luteal phase 200 to 800 mU oxytocin inhibited only human chorionic gonadotropin-stimulated progesterone output. Oxytocin did not affect luteal cell progesterone production in the midluteal phase. Thus oxytocin is present in corpora lutea, can be localized in the luteal cells, is probably produced locally, and may modulate luteal cell progesterone production.

Immunoreactive oxytocin is detectable in the corpora lutea of women and cynomolgus monkeys by radioimmunoassay. To localize the presence of oxytocin and neurophysin I in ovarian tissues of subhuman primates, three corpora lutea and ovarian stromal tissues and two Fallopian tubes obtained during the menstrual cycle of the baboon and decidua from two pregnant baboons were examined using highly specific antisera against either oxytocin or neurophysin I and peroxidase-antiperoxidase light microscopy immunohistochemistry. Oxytocin-like as well as neurophysin I-like immunoreactivities were found in some cells of all the corpora lutea only, but could not be demonstrated in ovarian stromal tissues, Fallopian tubes and decidua. Specificity of the immunocytochemical reaction was further confirmed by immunoabsorption of the antiserum with excess oxytocin or neurophysin, after which the immunoreactivities for both oxytocin and neurophysin in the luteal tissue were negative. Similar controls using normal rabbit serum gave no positive staining for either oxytocin or neurophysin. Counterstaining of the positive immunoreactivities for oxytocin and neurophysin I with Mayer's haematoxylin and eosin demonstrated clearly that the oxytocin and neurophysin I appeared as granular material mainly within the cytoplasm of the luteal cells. The localization of immunoreactive oxytocin and neurophysin I in the corpus luteum of the baboon demonstrates directly the presence of these two neurohypophysial peptides within primate luteal cells and suggests their local production.

To determine the site of oxytocin in human ovaries and its relationship with ovarian steroids, oxytocin and steroid hormones were measured in ovarian tissues, ovarian vein, and peripheral blood. Corpus luteum had significantly higher oxytocin, estrone, estradiol, and progesterone concentrations than corpus albicans and ovarian stroma (p = less than 0.01 to less than 0.001). Oxytocin concentrations in corpus luteum correlated significantly with estrone, estradiol, and progesterone. Oxytocin in corpus luteum increased from 14.0 +/- 1.8 ng/gm of wet weight in early to 30.8 +/- 0.9 ng/gm in midluteal phases (p = less than 0.001). Reverse phase high pressure liquid chromatography showed similarity between oxytocin in corpus luteum and synthetic oxytocin. Ovarian vein draining corpus luteum had significantly higher plasma oxytocin (11.8 +/- 1.5 pg/ml) than those without corpus luteum (2.1 +/- 0.2 pg/ml) or in the peripheral blood (2.9 +/- 0.3 pg/ml) (p = less than 0.001). Oxytocin in corpus luteum correlated significantly with its ipsilateral ovarian vein level of oxytocin, estrone, progesterone, and 17 alpha-hydroxyprogesterone. Our findings demonstrate that oxytocin is present and probably produced in corpus luteum and secreted into its ovarian vein; it may regulate corpus luteum release of progesterone, 17 alpha-hydroxyprogesterone, and estrone.

To assess fetal oxytocin release in relation to maternal oxytocin, serial paired maternal (femoral artery) and fetal (aorta) plasma oxytocin were determined by a specific and sensitive radioimmunoassay during pregnancy and parturition in the sheep. The maternal oxytocin level was 29.1 +/- 2.6 pg/ml (mean +/- SE) but was significantly lower than fetal oxytocin levels of 45.8 +/- 4.3 pg/ml (p less than 0.001). There was a significant correlation between paired maternal oxytocin and fetal oxytocin levels throughout late pregnancy (r = 0.2523, p less than 0.05). Maternal oxytocin levels calculated for every 5 days' gestation ranged from 15.8 +/- 2.7 to 32.7 +/- 5.6 pg/ml while the corresponding fetal oxytocin levels (21.6 +/- 3.5 to 62.5 +/- 21.6 pg/ml) were always higher (92 to 135 days) but not significantly different. However, at 136 to 142 days' gestation, fetal oxytocin levels (63.9 +/- 14.1 pg/ml) were significantly higher than maternal oxytocin levels (20.1 +/- 3.3 pg/ml) (p less than 0.01). Based on observations in two animals, fetal oxytocin levels appeared higher than maternal oxytocin levels at parturition with an increase a few days before labor. Our findings indicate that, in the sheep, fetal oxytocin levels were usually higher than but not significantly different from maternal oxytocin levels and that fetal oxytocin levels changed in relation to labor, suggestive of fetal release of oxytocin.

Recently, using a highly specific radioimmunoassay, we have demonstrated that the concentration of oxytocin in the corpus luteum of the human and cynomolgus monkey are several fold higher than in the peripheral circulation. In this study, we have examined the corpora lutea and ovarian stroma from the ovaries of normal adult cycling baboons (Papio anubis) for the presence of oxytocin through the use of immunocytochemical procedures. Tissues obtained at laparotomy were fixed in Bouin's solution and embedded in paraffin; immunoreactive oxytocin was localized with peroxidase-antiperoxidase and 3.3' diaminobenzidine. Six corpora lutea with stroma were obtained--two each from the early (Day 14-20), mid-(Day 21-24), and late (Day 25-30) stages of the luteal phase. Immunoreactive oxytocin was localized in all corpora lutea examined but was absent from all stroma samples. Larger areas of the corpus luteum from the mid-luteal phase showed staining for oxytocin, and the intensity of staining for this peptide was maximal in this phase of the cycle.

Serial plasma oxytocin (OT), PRL, TSH, FSH, LH, estrone, estradiol, and progesterone were measured by RIA in 12 women before and during a 30-min breast-feeding period on the third or fifth postpartum day. Plasma OT increased significantly from 10.8 +/- 3.4 to 22.4 +/- 3.5 pg/ml (mean +/- SE) within 2 min of suckling (P = less than 0.05) to reach the mean peak level of 53.2 pg/ml at 10 min. The increase in plasma OT was bimodal. Plasma PRL and TSH also increased significantly from baseline levels of 192 +/- 39 ng/ml and 16.9 +/- 5.6 microU/ml, respectively, to reach maximum levels of 427 +/- 91 ng PRL/ml at 10 min (P = less than 0.025) and 281.5 +/- 56.6 microU TSH/ml at 25 min (P = less than 0.005). Plasma FSH-beta (range of means, 3.5-4.6 ng/ml), LH (range of means, 1.7-2.6 mIU/ml), and estradiol (range of means, 29.8-38.2 pg/ml) were low and remained unchanged throughout breast feeding. Plasma progesterone was 6.0 +/- 0.4 ng/ml before breast feeding and did not alter significantly during breast feeding. The significance of these findings is discussed in relation to the milk let-down reflex and the relationship of TSH to PRL.

To determine if oxytocin (OT) is present in cynomolgus monkey corpus luteum, OT was measured by a specific and sensitive RIA in 13 corpora lutea, ovarian venous plasma on the ipsilateral side and peripheral venous plasma at different stages of the luteal phase. Serial dilution of acetic acid extract of the corpus luteum showed parallelism with standard OT in the RIA. Total content of OT in corpus luteum was 1.9 +/- 0.5 ng (mean +/- SEM) with a content of 0.4-0.8 ng in early luteal phase, 1.0-6.2 ng in midluteal phase, and 0.4-0.7 ng in late luteal phase. OT concentrations in corpus luteum were 21.0-75.2 ng/g wet wt in early luteal phase, increasing to 34.4-602.5 ng/g in midluteal phase; and declining to 3.4-117.4 ng/g in late luteal phase. OT concentrations per mg protein in the corpus luteum were 0.05-19.6 ng with peak concentrations of 14.7-19.6 ng/mg protein on day 22. Sephadex G-25 column chromatography of the corpus luteum extract revealed a single peak for binding activity similar to that of synthetic OT on the RIA. Ovarian vein blood from the same side as the corpus luteum had a significantly higher OT concentrations of 161.2 +/- 29.7 pg/ml on days 15-24 than 16.8 +/- 3.6 pg/ml on days 25-28 (P less than 0.01) and peripheral plasma OT levels of 23.2 +/- 3.4 pg/ml (P less than 0.025). Our findings indicate that OT is present and probably produced by monkey corpus luteum with peak OT concentrations found in midluteal phase. Thus OT may play a role in primate corpus luteum function.

To determine whether the human corpus luteum is a source of relaxin and oxytocin, we measured the concentrations of these peptides in plasma obtained from the ovarian veins of ovaries with and without a corpus luteum and compared these to peripheral plasma levels. Peripheral and ovarian venous blood samples were obtained from 34 nonpregnant women, 13 during the luteal phase and 21 during the follicular phase of their cycles, and from a 6-week pregnant woman. Plasma relaxin, oxytocin, and progesterone concentrations were determined by sensitive and specific RIAs. Plasma relaxin levels were not detectable (less than 0.16 microgram/L) in peripheral or ovarian venous plasma not draining a corpus luteum. The mean relaxin concentration in plasma draining an ovary with a corpus luteum was 0.41 +/- 0.09 (+/- SE) microgram/L. Oxytocin levels also were significantly higher in plasma draining an ovary with a corpus luteum (6.70 +/- 1.86 pmol/L) than in that draining the ovary with no corpus luteum (1.58 +/- 0.09 pmol/L; P less than 0.01) or in peripheral plasma (1.58 +/- 0.09 pmol/L; P less than 0.025). The mean progesterone concentration also was highest in plasma draining an ovary with a corpus luteum (210.2 +/- 50.5 nmol/L) compared with those in plasma from the contralateral ovarian vein (40.3 +/- 16.5 nmol/L P less than 0.005) and peripheral plasma (30.2 +/- 5.7 nmol/L; P less than 0.005) during the luteal phase. In a woman who was 6 weeks pregnant, plasma draining the ovary with a corpus luteum had 1.9 micrograms relaxin/L, but only 0.49 pmol/L oxytocin; the latter was similar to concentrations in noncorpus luteum-bearing ovarian venous plasma. These findings indicate that the human corpus luteum secretes relaxin, oxytocin, and progesterone. Both ovarian oxytocin and relaxin may function as paracrine or autocrine modulators of luteal function.

Objectives: This study was undertaken to determine expression levels of oxytocin receptor and its gene in peri-implantation phase human endometrium during clomiphene-treated cycles compared with control cycles.Study Design: Oxytocin receptor and its messenger ribonucleic acid in peri-implantation phase endometrium during control and clomiphene-treated (50 mg days 5 to 9) cycles of 5 healthy fertile women were determined by immunohistochemical methods, Western blot analysis with monoclonal antibody against amino acids 20 through 40 of the extracellular N-terminal human oxytocin receptor, and reverse transcription-polymerase chain reaction with oligonucleotide primers to amplify the 391-base pair fragment of the oxytocin receptor gene.Results: Oxytocin receptor and its messenger ribonucleic acid were expressed in human peri-implantation phase endometrial samples from both control and clomiphene-treated cycles. The receptor was localized predominantly in the epithelial cells and glands, with little or none detected in the stroma. Oxytocin receptor protein was separated out as a single 70-kd band by Western blot analysis; its relative abundance was significantly reduced during clomiphene-treated cycles. The messenger ribonucleic acid was detected in all endometrium during control and clomiphene-treated cycles, with greater expression during control cycles.Conclusions: The expressions of oxytocin receptor and its gene in luteal phase human endometrium suggest a functional relevance in modulation of biochemical changes for implantation. [ABSTRACT FROM AUTHOR]