Early Pregnancy In Rat Research Articles

A role for voltage‐gated, but not Ca2+‐activated, K+ channels in regulating spontaneous contractile activity in myometrium from virgin and pregnant rats

The roles of voltage-gated (K(V)) and large conductance Ca2+-activated K+ (BK(Ca)) channels in regulating basal contractility in myometrial smooth muscle are unresolved. The aim of this study was to determine the effects of inhibition of these channels on spontaneous rhythmic contraction in myometrial strips from four groups of rats: nonpregnant and during early (day 7), mid- (day 14), and late (day 21) pregnancy. BK(Ca) channels were inhibited using iberiotoxin (1-100 nM), paxilline (1-10 microM) or penitrem A (1-500, or 3000 nM); K(V) channels were inhibited using tetraethylammonium (TEA; 1-10 mM) and/or 4-aminopyridine (4-AP; 1-5 mM). Contractility was assessed as mean integral tension (MIT). Time/vehicle controls were also performed. None of the selective BK(Ca) channel inhibitors significantly affected contractility in myometrial strips from either nonpregnant or pregnant animals. 4-AP caused concentration-dependent increases in MIT in myometrium in all four groups. TEA (5 and 10 mM) significantly increased MIT in myometrium from nonpregnant, and mid- and late pregnant rats, but not in myometrium from early pregnant rats. TEA and 4-AP still caused an increase in MIT following treatment with 3000 nM penitrem A or a combination of propranolol, phentolamine, atropine (all 1 microM) and capsaicin (10 microM) in myometrial strips from nonpregnant rats. These results indicate that whereas BK(Ca) channels play little or no part in controlling basal rhythmicity in rat myometrium, K(V) channels appear to play a crucial role in this regard, especially during mid- and late pregnancy.

Expression of nidogens in rat uterus and embryo during decidualization and implantation

The purpose of this study was to demonstrate the expression of nidogen-1 and nidogen-2 and their possible role in decidualization and implantation events during early pregnancy in rats. The tissue samples were examined from pregnant animals between gestational days 1-8 using immunocytochemistry. The uterine luminal epithelium, the glandular epithelium, and the myometrial smooth muscle cells stained strongly from gestational days 1-8 with both nidogen antibodies. At day 4 the decidual reaction areas began to appear in the stromal matrix and immunostaining of both nidogens revealed that the basement membrane of the surface epithelium was discontinuous. The differentiation of stromal cells into decidual cells was seen at gestational day 5 and both nidogens were weakly expressed in the decidualizing cells. At day 6, nidogen-2 immunoreactivity was higher in the primary decidual cells close to the embryo than nidogen-1, and during development of the decidual tissue both nidogens appeared in the endometrial stromal cells. At day 7, while expression of both nidogens declined in the primary decidual cells, their expression was markedly observed in the secondary decidual cells close to the myometrium. At day 8, expression of both nidogens was also observed to increase in the primary decidual cells. While nidogen-2 expression was seen in the parietal endoderm and primary ectoderm of the rat embryos at this developmental stage, nidogen-1 expression was only detected in the parietal endoderm. These results indicate that nidogen-1 and nidogen-2 could play important roles during embryogenesis, decidualization, and implantation in the endometrium of rat uterus.

RNA fingerprinting using RAP-PCR identifies an EBAF homologue mRNA differentially expressed in rat oviduct

As a step towards the identification of genes preferentially expressed in the oviduct during early rat embryo development, we isolated a cDNA fragment (Pr14) by using RNA arbitrarily primed PCR (RAP-PCR), being its expression restricted to oviduct and uterus; its mRNA is mainly expressed in oviduct during late luteal phase and early pregnancy. This fragment is 100% identical to a rat DNA sequence (Accession No. NW_047400) downstream the terminal exon of a Ratturs norvegicus gene (Locus Link Accession No. LOC289316) similar to ebaf (endometrial bleeding-associated factor), a novel member of the Transforming Growth Factor superfamily. Northern analyses showed that this sequence hybridizes with 2.9 kb and 4.1 kb mRNAs in early pregnant rat oviducts. However, only the 4.1 kb mRNA was detected in the oviduct of non-pregnant rats, showing an increase from proestrus to diestrus. The expression of this oviduct-uterus specific mRNA suggests that the products of this gene may play a role in the oviductal reproductive process.

Nitric oxide induces gelatinase A (matrix metalloproteinase 2) during rat embryo implantation

Objective To evaluate a reciprocal signaling interaction initiated by embryo-derived nitric oxide (NO) to facilitate implantation by increased production of gelatinase A (matrix metalloproteinase 2, MMP2) in uterine stroma. Design Experimental animal studies. Setting Reproductive-physiology research laboratory. Animal(s) Female syngeneic Wistar rats aged 14 weeks. Intervention(s) Vaginal smears to confirm pregnancy. Oviductal ligature to avoid the descent of blastocysts to the uterine lumen. Plasma exudation assays to locate uterine blastocyst implantation sites. Organ cultures treated with NO donors and nitric oxide synthase (NOS) inhibitors. Main outcome measure(s) Expression of MMP2 and NO was assessed by Western blot and zymography of tissue extracts and by immunofluorescence of tissue sections. Result(s) An increase in MMP2 activity was found in uterine extracts in early pregnant rats and was concentrated at implantation sites. Immunolocalization experiments showed that inducible NOS was expressed on the surface of the implanting blastocyst adjacent to the uterine epithelium at the sites of increased MMP2 expression. In organ culture experiments, NO donors were found to increase, whereas NOS inhibitors were found to decrease MMP2 activity in uterine tissue sections. Conclusion(s) Blastocyst-derived NO contributes to the production of uterine-derived MMP2, an essential component of implantation and initiation of placentation.

Expression of MAPkinases (Erk1/2) during decidualization in the rat: regulation by progesterone and nitric oxide.

The interaction between nitric oxide (NO), progesterone and the MAPkinase signalling pathway involved in decidualization was studied using immunohistochemistry during implantation in the rat. Early pregnant rats were treated with the inhibitor of nitric oxide synthesizing enzyme iNOS, aminoguanidine, either alone or in combination with the low dose antiprogestin, onapristone. The combined treatment was most effective on days 7 and 9 post coitum leading to a complete loss of embryos. The expression pattern of activated MAPkinases, Erk1/2 and iNOS appeared to be associated with the differentiation process of decidualization. A maximum staining of both enzymes was observed on day 9 post coitum in the mesometrial decidua. In addition, Erk1/2 and iNOS were highly coexpressed around the mesometrial sinusoids. Combined treatment with aminoguanidine and onapristone for 3 days led to a transient suppression of Erk1/2 and abolished Cox2 expression. Concomitantly, angiogenesis was reduced and dilated sinusoids were missing in the mesometrial decidua. In conclusion, our study suggests that (i) the member of the mitogen-activated protein kinase (MAPK) family, Erk1/2, is activated during implantation and may play an important role during the decidualization process, and (ii) this enzyme may be regulated by both progesterone and NO.

Effects of Annona squamosa extract on early pregnancy in rats.

Annona squamosa Linn., family Annonaceae, is said to show varied medicinal effects, including insecticide, antiovulatory and abortifacient. The purpose of present study was to investigate if A. squamosa seed aqueous extract, in doses higher than that popularly used to provoke abortion, interferes with reproductive performance, and to correlate the ingestion of this extract with possible alterations in rat embryonic implantation. Doses of 300 mg/kg (Treated Group I, n = 17) and 600 mg/kg (Treated Group II, n = 12) body wt. were administered by gavage, during days 1 to 5 of pregnancy (preimplantation period). The control group (n = 13) received water in the same manner, during the same period for comparison with experimental groups. The animals were euthanized on day 10 of pregnancy. Treatment of dams during the preimplantation period showed no signs of toxicity, and no alteration in the corpora lutea, implantations and embryo in terms of development numbers. The percentage of preimplantation and postimplantation losses in treated groups I and II did not differ from those of control. Treatment with aqueous extract of A. squamosa seeds caused no morphological change in the endometrium. The absence of morphological alterations in uterine epithelial cells in treated groups I and II permitted a viable embryonic implantation, as verified by the number of embryos in development at day 10 of pregnancy. Thus, A. squamosa seed aqueous extract did not interfere with the reproductive performance of pregnant rats.

Toxic effects of butyltin trichloride during early pregnancy in rats

The objective of this study was to determine the toxic effects of butyltin trichloride (BTCl) during early pregnancy. Following successful mating, female rats were given BTCl by gastric intubation at 0, 56, 226, or 903 mg/kg on days 0–3 or 4–7 of pregnancy. Female rats were sacrificed on day 20 of pregnancy and fetuses were examined for number, abnormality, mortality, and weight. The maternal body weight gain and food consumption during the administration period was significantly decreased after administration of BTCl at 903 mg/kg on days 0–3 or 4–7 of pregnancy. The pregnancy rate in the BTCl-treated groups was comparable to the control value, regardless of the days on which BTCl was given. The incidence of pre-implantation embryonic loss was not significantly affected after administration of BTCl on days 0–3 or 4–7. In females having implantations, the numbers of corpora lutea, implantations, and live fetuses and the incidences of pre- and postimplantation loss in the groups given BTCl on days 0–3 were comparable to the controls. Although a significant increase in the incidence of postimplantation loss was observed after administration of BTCl on days 4–7 at 56 mg/kg, this change was small and inconsistent across doses and seems unlikely to be toxicologically significant. A significant decrease in weight of female fetuses was found after administration of BTCl at 903 mg/kg on days 0–3 or 4–7. It could be concluded that BTCl treatment during early pregnancy is maternal and developmental toxic at 903 mg/kg.

Characterization of fibroblastic cell plasticity in the lamina propria of the rat uterine cervix at term.

Different organs contain fibroblasts with specific features and functions, indicating the complexity of fibroblast biology. In the rat cervical stroma, fibroblasts are preferentially located in the fibrous ring that surrounds the mucous layer. The purpose of this study was to investigate the morphological features and immunophenotype of fibroblastic cells of the uterine cervix in cycling, pregnant, and postpartum rats. Expression of the cytoskeletal proteins desmin, vimentin, and alpha-smooth muscle actin (alpha-SMA) were studied by immunohistochemistry. The optical density of immunohistochemical staining was quantified by image analysis. The ultrastructural features of fibroblastic cells were observed under transmission electron microscopy. Cervical fibroblastic cells always expressed vimentin and desmin but never alpha-SMA. During the first half of pregnancy (Day 5 [D5] to D14), desmin intensity values were similar to those of cycling and postpartum fibroblasts. In contrast, a strong expression of desmin was found from D15 to D22, with maximal expression at term (D23). Immunohistochemical expression for vimentin was constant throughout pregnancy and showed no differences with cycling and postpartum uterine cervices. Stromal cells from cycling and early pregnant rats displayed ultrastructural features characteristic of typical fibroblasts. In contrast, at the end of pregnancy, fibroblasts differentiated and showed increased secretory characteristics, reaching the ultrastructural features of a myofibroblast. Based on the differential expression of desmin and the electron microscopic observations, the foregoing results showed a modulation of the fibroblastic phenotype in the uterine cervix during pregnancy. To our knowledge, this is the first report that addresses the presence of myofibroblasts derived from resident fibroblasts in the fibrous ring of the rat uterine cervix. Fibroblastic-myofibroblastic cell plasticity may have implications in the physiological changes displayed in the uterine cervix during pregnancy, parturition, and postpartum involution.

Conditioned medium from hypoxic cytotrophoblasts alters arterial function

Objective: Our goal was to test the hypothesis that cytotrophoblasts, under low oxygen tension, release substances that affect vascular behavior. Study Design: We studied the vascular response to the vasoconstrictors phenylephrine (receptor dependent) and potassium (receptor independent), the relaxation response to methacholine, and the vasomotor behavior of isolated resistance (mesenteric) arteries from early pregnant rats after incubation in conditioned medium from first-trimester cytotrophoblasts, maintained in standard or hypoxic (2%; 14 mm Hg) culture conditions. Results: After incubation in medium from hypoxic cytotrophoblasts, arterial segments were more responsive to phenylephrine and to potassium-induced constriction but were less responsive to methacholine, and the vasomotor activity was increased compared with that found in vessels incubated in control medium. Conclusions: These changes in vascular behavior are similar to those reported in isolated arteries from women with preeclampsia. These studies provide evidence which suggests that the link between abnormal placentation and maternal vascular abnormality in preeclampsia is the elaboration of vasoactive factors by cytotrophoblasts in response to hypoxia. (Am J Obstet Gynecol 2001;184:984-90.)

Effects of 4- tert-octylphenol on initiation and maintenance of pregnancy following oral administration during early pregnancy in rats

4- tert-Octylphenol (OP) is an alkylphenol that is an intermediate in the production of alkylphenol ethoxylates. OP has been reported to be the most potent estrogenic alkylphenol in vitro. In the present study, the effects of OP on initiation and maintenance of pregnancy were investigated in rats. Inseminated female rats were orally given OP at 0, 15.6, 31.3, 62.5 and 125 mg/kg on day 0 through day 8 of pregnancy. Female rats were sacrificed on day 20 of pregnancy, and pregnancy outcome was determined. Decreases in body weight gain and food consumption on days 0–9 were found at 31.3 mg/kg and above, and at 15.6 mg/kg and above, respectively. The pregnancy rate was not adversely affected by OP administration during early pregnancy even at 125 mg/kg. The incidence of post-implantation loss per litter at 31.3 mg/kg and above was significantly higher than that in the control group. The body weights of live fetuses in the OP-treated groups were not significantly different from those in the control group. No increase in the incidence of fetuses with external malformations was found in any OP-treated group. We concluded that OP during early pregnancy caused post-implantation embryonic loss at doses that showed maternal toxicity.

Adverse effects of dibutyltin dichloride on initiation and maintenance of rat pregnancy

The present study was conducted to evaluate the adverse effects of dibutyltin dichloride (DBTCl) on initiation and maintenance of pregnancy after maternal exposure during early pregnancy in rats. After successful mating, female rats were given DBTCl by gastric intubation on Days 0 to 3 or on Days 4 to 7 of pregnancy at 0, 3.8, 7.6, or 15.2 mg/kg. Food-restricted pregnant rats were given an amount of feed equal to the feed intake of female rats treated with DBTCl at 15.2 mg/kg on Days 0 to 3 or on Days 4 to 7 of pregnancy. Female rats were sacrificed on Day 20 of pregnancy and pregnancy outcome was determined. After administration of DBTCl on Days 0 to 3, the rate of nonpregnant females and the incidence of preimplantation embryonic loss in the 7.6 mg/kg group were significantly higher than those in the control group, and those in the 15.2 mg/kg group were significantly higher than those in the control and pair-fed groups. In females with implantations, the numbers of implantations and live fetuses and the incidence of postimplantation embryonic loss in the groups given DBTCl on Days 0 to 3 were not significantly different from those in the control group. The incidence of postimplantation embryonic loss in the groups given DBTCl on Days 4 to 7 at 7.6 and 15.2 mg/kg was significantly higher than that in the control and pair-fed groups. It can be concluded that DBTCl adversely affects initiation and maintenance of pregnancy when administered during early pregnancy and that the manifestations of the adverse effects of DBTCl vary with the gestational stage at the time of maternal exposure.

Potentiation of the hypotensive effect of adrenomedullin in pregnant rats

The hypotensive effect of adrenomedullin, a potent vasodilator peptide, was examined in conscious pregnant (6, 13 and 20 days of pregnancy) and non-pregnant rats. The intravenous administration of adrenomedullin (0.01–3.0 nmol/kg) produced a dose-dependent depressor response in pregnant and non-pregnant rats. At low doses (0.01–0.1 nmol/kg), the maximum decrease in blood pressure was significantly higher in pregnant rats (20 days pregnant) than in non-pregnant rats. At high doses, no significant difference was observed between the two groups. Furthermore, the administration of adrenomedullin did not significantly affect the basal mean blood pressure (MBP) at any dose when compared to the non-pregnant group at 6 and 13 days of pregnancy. In the ovariectomized rats, the depressor responses in 17β-estradiol-treated, progesterone-treated and 17β-estradiol+progesterone-treated rats were not significantly different from that in the control rats, suggesting that the augmented effect on the depressor response to adrenomedullin in pregnant rats may not be due to hormonal changes during pregnancy. The adrenomedullin receptor mRNA level of the descending thoracic aorta was significantly higher in the late-pregnancy rats (20 days of pregnancy). However, the levels did not show any difference between the early-pregnant rats (6 and 13 days of pregnancy) and the non-pregnant rats. These findings suggested that the changes in the depressor response to adrenomedullin which occur at term in pregnant rats may be mediated by changes of adrenomedullin receptor gene expression rather than by sex hormones.

Dibromoacetic acid does not adversely affect early pregnancy in rats

Dibromoacetic acid (DBA), a byproduct of water disinfection, has been shown to have adverse reproductive effects in male rodents. In light of the lack of data on potential effects on female reproduction, studies were initiated to evaluate the potential effect of the chemical on early pregnancy. Groups of mature Holtzman rats were used, and cycles were monitored in all animals by vaginal smears. The administration of DBA at 0, 62.5, 125, or 250 mg/kg/d to rats during the first 8 d of pregnancy had no effect on the number of implantation sites found on Day 9 nor any other progestational parameter, except for serum estradiol, which was elevated. When groups of pregnant rats treated with the same range of dosages of DBA during Days 1 through 8 of pregnancy were killed on Day 20, there was no effect of treatment on the number of pups/litter, number of resorptions, or mean pup weight. These data demonstrate that while DBA has serious adverse effects on reproduction in male rodents, the chemical does not affect early pregnancy in rats. Effects on ovarian function and latent fertility are not ruled out.

Mechanism of volume adaptation in the awake early pregnant rat.

The objective of the present study was to determine whether the increase in plasma volume (PV) during pregnancy is established by fluid retention or by a shift within the extracellular fluid volume (ECFV) from the interstitium toward the intravascular compartment. To this end, we simultaneously, measured total body water, (TBW), ECFV, and PV together with the hematocrit (Hct) and plasma osmolality 4, 8, and 12 days postsurgery in chronically instrumented pregnant (P) and nonpregnant (NP) rats. The P rats were instrumented with a catheter in the femoral artery on day 1 postconception. In the NP group, neither TBW nor ECFV and PV had changed consistently on days 8 and 12 postsurgery relative to day 4. In contrast, in the P animals, TBW, ECFV, and PV had increased by 16, 24, and 20%, respectively, by day 12 relative to day 4. To evaluate whether PV had increased in concert with an overall rise in TBW or as a result of a fluid shift at the cost of the interstitial fluid volume, we calculated the relative size of each fluid compartment on three consecutive measurement sessions. In the NP group, TBW, presented as percentage of maternal weight (%MW) as well as ECFV (%TBW) and PV (%ECFV) had not changed consistently throughout the measurement period. In the P animals, TBW (%MW) was slightly higher on day 12 compared with day 4, but ECFV (%TBW) and PV (%ECFV) had not changed significantly. Finally, in the NP group, Hct had not changed, whereas, in the P animals, Hct was 10% lower on days 8 and 12 compared with day 4. Plasma osmolality did not change consistently in either group during the course of the experimental period. The gradual synchronous increase in all fluid compartments, without consistent change in their relative distribution, suggests that, in normal rat pregnancy, PV expansion is primarily achieved by fluid retention rather than by a redistribution of the ECFV.

Attenuated neuroendocrine responses to emotional and physical stressors in pregnant rats involve adenohypophysial changes.

1. The responsiveness of the rat hypothalamo-pituitary-adrenal (HPA) axis and hypothalamo-neurohypophysial system (HNS) to emotional (elevated plus-maze) and physical (forced swimming) stressors and to administration of synthetic corticotrophin-releasing hormone (CRH) was investigated during pregnancy and lactation. In addition to pregnancy-related adaptations at the adenohypophysial level, behavioural responses accompanying the neuroendocrine changes were studied. 2. Whereas basal (a.m.) plasma corticosterone, but not corticotrophin (adrenocorticotrophic hormone; ACTH), levels were increased on the last day (i.e. on day 22) of pregnancy, the stress-induced rise in both plasma hormone concentrations was increasingly attenuated with the progression of pregnancy beginning on day 15 and reaching a minimum on day 21 compared with virgin control rats. A similar attenuation of responses to both emotional and physical stressors was found in lactating rats. 3. Although the basal plasma oxytocin concentration was elevated in late pregnancy, the stress-induced rise in oxytocin secretion was slightly lower in day 21 pregnant rats. In contrast to vasopressin, oxytocin secretion was increased by forced swimming in virgin and early pregnant rats indicating a differential stress response of these neurohypophysial hormones. 4. The blunted HPA response to stressful stimuli is partly due to alterations at the level of corticotrophs in the adenohypophysis, as ACTH secretion in response to CRH in vivo (40 ng kg-1, i.v.) was reduced with the progression of pregnancy and during lactation. In vitro measurement of cAMP levels in pituitary segments demonstrated reduced basal levels of cAMP and a lower increase after CRH stimulation (10 nM, 10 min) in day 21 pregnant compared with virgin rats, further indicating reduced corticotroph responsiveness to CRH in pregnancy. 5. The reduced pituitary response to CRH in late pregnancy is likely to be a consequence of a reduction in CRH receptor binding as revealed by receptor autoradiography. [125I] CRH binding in the anterior pituitary was significantly reduced in day 11, 17 and 22 pregnant rats compared with virgin controls. 6. Anxiety-related behaviour of the animals as revealed by the time on and entries into the open arms of the elevated plus-maze was different between virgin and pregnant rats with decreased number of entries indicating increased anxiety with the progression of pregnancy (except on pregnancy day 18). The emotional behaviour, however, was not correlated with the neuroendocrine responses. 7. The results indicate that the reduced response of the HPA axis to stressors described previously during lactation is already manifested around day 15 of pregnancy in the rat and involves physiological adaptations at the adenohypophysial level. However, alterations in stressor perception at higher brain levels with the progression of pregnancy may also be involved.

Blood pressure and heart rate variability in early pregnancy in rats.

Changes in the autonomic control of the circulation may contribute to the maternal hemodynamic adaptation to early pregnancy. To evaluate this, we studied mean arterial pressure (MAP) and heart rate (HR) in chronically instrumented, conscious rats in early (days 4, 6, 8, and 10) and late (day 18) pregnancy (n = 8) and in nonpregnant rats (n = 9). MAP and HR were recorded on a beat-to-beat basis and analyzed by spectral analysis. Spectral density power was calculated in low- (0.047-0.305 Hz), mid- (0.305-0.598 Hz), and high-frequency (0.598-1.494 Hz) bands, which contain oscillations that are among others related to myogenic-, sympathetic/vagal-, and vagal/respiration-related influences, respectively. In addition, baroreceptor reflex sensitivity was determined from spontaneous variations in MAP and HR by a sequential time series method and by calculating the transfer gain between MAP and HR in the midfrequency band. Mean values of HR and MAP did not differ between the two groups on day 4. In the pregnant group, MAP fell gradually over days, whereas HR had significantly increased only on day 18. Overall variability in MAP and HR (expressed as coefficients of variation) did not change during pregnancy. Baroreceptor reflex sensitivity did not differ between the groups and did not change with advancing pregnancy. Spontaneous oscillations of MAP and HR at low, mid, and high frequencies were not different between pregnant and nonpregnant rats on days 4 to 10. On day 18, spectral density power of MAP, but not of HR, in the high-frequency band had significantly increased in pregnant rats only, most likely reflecting the increased impact of breathing on MAP fluctuations. We conclude that, with the methods employed, we could not discern any changes in baroreflex sensitivity and MAP and HR variability in pregnancy. This would imply that changes in autonomic activity do not contribute appreciably to the hemodynamic adaptations in early rat pregnancy.

Embryolethality following maternal exposure to dibutyl phthalate during early pregnancy in rats.

Embryolethality following maternal exposure to dibutyl phthalate during early pregnancy in rats.

The relaxation responses to corticotropin-releasing factor in rat aorta are endothelium dependent and gestationally regulated

OBJECTIVE: Our purpose was to examine the hypothesis that the relaxant response to corticotropin-releasing factor is endothelium dependent and changes during pregnancy. STUDY DESIGN: Relaxant responses to cumulative concentrations of corticotropin-releasing hormone were measured in rings of thoracic aorta, precontracted with phenylephrine, from pregnant and nonpregnant female rats. Each group consisted of 5 to 10 rats. RESULTS: The relaxation induced by corticotropin-releasing factor in aortic rings from nonpregnant rats decreased after deendothelization and treatment with Nω-nitro-l-arginine methyl ester (a nitric oxide synthase inhibitor), LY-83,583 (a guanylate cyclase inhibitor), or α-helical corticotropin-releasing factor 9-41 (a corticotropin-releasing factor antagonist). The percent decrease in relaxation at 1 nmol/L of corticotropin-releasing factor was about 88% with deendothelization, 100% with Nω-nitro-l-arginine methyl ester, 76% with LY-83,583, and 100% with α-helical corticotropin-releasing factor 9-41. The responses to corticotropin-releasing factor in intact rings from rats in early pregnancy were not significantly different from those in the nonpregnant female rats, but the responses were decreased during the later stages of gestation (percent decrease in relaxation at 1 nmol/L of corticotropin-releasing factor about 71% at day 20 and 98% at term). The relaxation induced by corticotropin-releasing factor during pregnancy was also inhibited by deendothelization, Nω-nitro-l-arginine methyl ester, or LY-83,583. CONCLUSIONS: The relaxant response to corticotropin-releasing factor in the rat aorta is endothelium dependent and is mediated by the nitric oxide–cyclic guanosine monophosphate pathway. The receptor involved in this effect is α-helical corticotropin-releasing factor 9-41 sensitive. This inhibitory response is unchanged during early pregnancy but reduced toward the end of gestation. (Am J Obstet Gynecol 1997;176:234-40.)

Sialomucin Complex, a Heterodimeric Glycoprotein Complex: EXPRESSION AS A SOLUBLE, SECRETABLE FORM IN LACTATING MAMMARY GLAND AND COLON

Ascites 13762 rat mammary adenocarcinoma cells express abundantly on their cell surfaces a heterodimeric glycoprotein complex composed of a sialomucin ascites sialoglycoprotein (ASGP)-1 and a transmembrane subunit ASGP-2. The latter, which contains two epidermal growth factor-like domains, binds the receptor tyrosine kinase p185(neu), suggesting that the complex is bifunctional as well as heterodimeric. Immunoblot analyses using monoclonal antibodies prepared against the complex demonstrate high levels of expression in rat lactating mammary gland and colon. Immunolocalization studies with anti-ASGP-2 indicate that ASGP-2 is present in these two tissues in the apical regions of secretory epithelial cells. Both mammary gland and colon contain a soluble, secretable form of ASGP-2, which is not found in the ascites cells; milk and mammary gland also have the membrane form. Immunoblot analyses using a COOH-terminal-specific polyclonal antibody indicate that the soluble form of ASGP-2 is missing its COOH-terminal domains. Both the soluble and membrane forms of ASGP-2 are similar to the membrane-associated form from the 13762 adenocarcinoma with respect to Mr, antigenicity, and association with ASGP-1. The presence of ASGP-1 in milk suggests that it is a candidate for the uncharacterized high Mr milk mucin, MUCX. ASGP-2 expression is up-regulated in mammary gland during pregnancy, because it is undetectable in virgin and early pregnant rats but abundant in the gland from late pregnant and lactating animals. However, compared with the lactating mammary gland, the 13762 ascites cells overexpress ASGP-2 by more than 100-fold, which may contribute to their malignancy. These combined results indicate that sialomucin complex is a unique secreted product in the mammary gland and colon, whose behavior is different from that in the mammary ascites tumors, and which may play important roles in mammary and intestinal physiology.

Gene expression of gonadotropin-releasing hormone in early pregnant rat and steroid hormone exposed mouse uteri.

While gonadotropin-releasing hormone (GnRH) or GnRH-like substance have been reported to exist in nonhypothalamic tissues such as placenta, gonads, and mammary gland, there have been no reports concerning the detection of GnRH mRNA in uterine tissue. In order to investigate the presence of GnRH in decidual tissues and its possible involvement in the regulation of placental function, we examined the gene for GnRH in the rodent uterus in early pregnancy and in nonpregnant animals treated with female sex steroids. Using RT-PCR and in situ hybridization we found GnRH mRNA transcripts in the rat uterus of 3rd and 6th day gestation and in the mouse uterus treated with estrogen and progesterone. In situ hybridization revealed that GnRH mRNA was localized in the endometrial stromal cells of the 3rd and 6th day of gestation. These results suggest the existence of GnRH gene expression in uterine stromal cells and its possible paracrine effect derived from the decidual cells.