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a Department of Molecular and Integrative Physiology
b and College of Medicine, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801
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ABSTRACT |
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 TOP ABSTRACT INTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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INTRODUCTION |
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TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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Small amounts of relaxin H2, and perhaps small amounts of relaxin H1 as well, are produced in human decidua, placenta, and mammary gland [3, 21, 22]. Bryant-Greenwood and coworkers [23, 24] provided evidence that decidual and placental relaxin has local autocrine and/or paracrine roles that contribute to connective tissue remodeling at the maternal-fetal interface during late pregnancy and at parturition. The addition of synthetic human relaxin H2 to fetal membrane explants caused a dose-related increase in the expression of the genes, proteins, and enzyme activities of the matrix metalloproteinases interstitial collagenase (MMP-1), stromelysin (MMP-3), and gelatinase B (MMP-9) [23, 24].
A major step toward encouraging the view that relaxin has physiological and/or therapeutic roles in the human would be the demonstration that human tissues contain cells with specific relaxin-binding sites, i.e., putative relaxin receptors. There is limited evidence that human tissues contain relaxin-binding sites. 32P-Labeled human relaxin was reported to bind with specificity to both the decidua and cytotrophoblast layers in fetal membranes [23] and to a human uterine cell line [25]. Also, biotinylated human relaxin was reported to bind to the human cervix [26]. None of these studies [23, 25, 26] identified the cells to which relaxin bound.
The objective of this study was to identify cells that contain specific and saturable binding sites for relaxin within several tissues that are potential targets for human relaxin, namely, cervix, vagina, uterus, mammary glands, mammary nipples, and placenta. To accomplish that objective, an in situ histochemical binding procedure recently developed by Min and Sherwood [27] for identification of relaxin-binding cells in the pig was employed.
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MATERIALS AND METHODS |
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TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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Human tissue samples were obtained immediately after clinical procedures. Uterine cervix, vagina, and uterus were obtained from hysterectomy specimens from six patients ranging from 29 to 51 yr of age. Five of the patients were premenopausal. Indications for hysterectomy were uterine fibroids (n = 3), adenomyosis with uterine prolapse (n = 2), and uterine leiomyomata (n = 1). Uterine cervix was also obtained from one woman at cesarean section. Mammary glands and mammary nipples were obtained after modified radical mastectomy, from three patients ranging from 56 to 69 yr of age, mean 64 yr. The indication for radical mastectomy was the presence of neoplasm. Grossly normal tissues were provided by the Pathology Department at Covenant Medical Center, Urbana, IL. Placenta was obtained after normal delivery by two women with uncomplicated pregnancy. Before participation, informed consent was obtained from all subjects. This study was approved by the University of Illinois Institutional Review Board and the Covenant Medical Center Investigational Review Committee.
Each tissue sample was cut into slices (0.5–3 cm3) and individually placed in peel-A-way plastic embedding molds (Polysciences, Inc., Warrington, PA). The tissues were frozen with Tissue-Tek O.C.T. embedding compound (Miles Scientific, Elkhart, IN) in liquid nitrogen and stored at -70°C until sectioning. Frozen sections (8 µm) were cut on an HR Mark II cryostat (Slee Medical Equipment Ltd., London, England) at -20°C and thaw-mounted on microscope slides coated with 0.2% poly-L-lysine (Mr 300 000). Tissue slides were incubated in anhydrous acetone at -30°C for 3 h.
Preparation and Characterization of Biotinylated Relaxin
Porcine relaxin was isolated as described by Sherwood and O'Byrne [28] and biotinylated by a modification [27] of the procedure described by Büllesbach and Schwabe [29]. In brief, porcine relaxin was dissolved in 0.2 M N-methylmorpholine-HCl buffer (pH 7.5) at a final concentration of 2 µmol/ml. To supply the biotinylating reagent in excess, 10 molar equivalents of biotinyl-
-aminocaproic acid-N-hydroxy succinimide ester (Sigma, St. Louis, MO) in dimethylformamide at a concentration of 100 µmol/ml were added to the relaxin. The reaction mixture was stirred at room temperature for 4 h, and the reaction was stopped by the addition of acetic acid until a 1 M acetic acid solution was obtained. The contents of the reaction mixture were separated from the biotinyl-
-amino hexanoyl-relaxin (biotinylated relaxin) by ultrafiltration using an Amicon model 402 stirred ultrafiltration apparatus with a Diaflo Ultrafilter type YM1 membrane (molecular weight cut-off 1000; Amicon, Beverly, MA). The N-methylmorpholine-HCl buffer and acetic acid were replaced with PBS (0.01 M NaH2 PO4 and 0.15 M NaCl, pH 7.4) in the ultrafiltration unit. The biotinylated relaxin was stored at a final concentration of 9 nmol/ml at -70°C.
The mean number of biotin molecules per biotinylated relaxin molecule was found to be 3.5 when determined by a spectrophotometric 4'-hydroxyazobenzene-2-carboxylic acid (HABA) assay according to the manufacturer's protocol (Pierce Chemical Co., Rockford, IL) [27]. The bioactivity of biotinylated relaxin was determined with a mouse interpubic ligament bioassay [30], and it did not differ statistically from that of unmodified porcine relaxin [27].
Immunohistochemical Localization of Biotinylated Relaxin
Immunohistochemical localization of biotinylated relaxin followed the procedure of Min and Sherwood [27]. In brief, the tissue slides were brought to room temperature, and subsequent immunohistochemical procedures were performed at room temperature. Tissue slides were incubated for 30 min in 50 mM glycine in PBS (pH 7.3) and then incubated for 3 h with blocking buffer 1 (1% BSA fraction V, 0.2% fish gelatin [BBInternational, Cardiff, UK], 5% normal pig serum, and 2 mM NaN3 in PBS). Tissue slides were incubated for 4 h in incubation buffer 1 (1% BSA fraction V, 0.2% fish gelatin, 1% normal pig serum, and 2 mM NaN3 in PBS) in four different ways. The first treatment incubated each tissue with biotinylated relaxin probe (4 µg/ml) in order to localize relaxin-binding sites. The second treatment incubated each tissue with unmodified porcine relaxin (4 µg/ml). Since the unmodified relaxin does not contain biotin, this treatment was used as a negative control. The third treatment incubated each tissue with biotinylated relaxin plus a 2000-fold excess of recombinant human insulin (Humulin R; Eli Lilly, Indianapolis, IN) in order to determine hormonal specificity of binding of the biotinylated relaxin probe. The fourth treatment incubated each tissue with biotinylated relaxin plus a 2000-fold excess of porcine relaxin [28] in order to determine whether there are finite numbers of relaxin binding sites in the tissue.
After incubation, tissue slides were rinsed for 2 h with five changes of wash buffer (1% BSA fraction V, 0.2% fish gelatin, and 2 mM NaN3 in PBS). The tissues were then fixed for 30 min with 3% glutaraldehyde in PBS, rinsed for 30 min in PBS, and incubated for 30 min with 50 mM glycine in PBS. The tissues were then incubated for 4 h in blocking buffer 2 (1% BSA fraction V, 0.2% fish gelatin, 5% normal goat serum, and 2 mM NaN3 in PBS), and for 4 more hours in 800 µl antibiotin IgG conjugated to 1 nm colloidal gold (BBInternational) diluted 1:25 with incubation buffer 2 (1% BSA fraction V, 0.2% fish gelatin, 1% normal goat serum, and 2 mM NaN3 in PBS). The tissues were rinsed for 2 h with 5 changes of wash buffer and postfixed with 3% glutaraldehyde in PBS for 30 min. All slides were rinsed in copious amounts of double-distilled water for 30 min before silver intensification of the gold particles. Silver intensification was performed by incubating slides in silver solution (BBInternational) for 10 min at room temperature. After being rinsed in copious amounts of double-distilled water for 10 min, the tissue sections were dehydrated in an ascending series of ethanol, cleared in Clear-Rite 3 (Richard Allen, Richland, MI), and coverslipped using mounting medium (Richard Allen).
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RESULTS |
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TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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Immunohistochemistry results from sections of the vagina are shown in Figure 2. Prominent labeling of luminal epithelial cells and both circular and longitudinal smooth muscle cells of the vagina was observed in sections incubated with biotinylated relaxin (Fig. 2, A–C). At the higher magnification, labeling was associated with smooth muscle surrounding blood vessels (Fig. 2, B and C). Relaxin binding also appeared to be associated with individual cells located within the stroma (Fig. 2, B and C). These are probably, at least in part, fibroblasts. No signal was detected in vaginal sections incubated with unmodified porcine relaxin (Fig. 2D). Binding of the biotinylated relaxin to the vagina was hormone-specific and saturable (Fig. 2, E and F).
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Immunohistochemistry results from sections of the uterus are shown in Figure 3. Prominent labeling of both luminal and glandular epithelial cells and myometrial smooth muscle cells was observed in sections incubated with biotinylated relaxin (Fig. 3, A and B). Labeling of less intensity was also observed in blood vessels (Fig. 3A). With the higher magnification of the border between the endometrium and myometrium, binding appeared to be associated with individual cells in the stromal extracellular matrix (Fig. 3B). No signal was detected in sections incubated with unmodified porcine relaxin (Fig. 3C). Binding of the biotinylated relaxin in the uterus was hormone-specific and saturable (Fig. 3, D and E).
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Immunohistochemistry results from sections of the mammary glands are shown in Figure 4. Prominent labeling of epithelial cells in both the lobular alveolar structures and lactiferous ducts as well as in blood vessels was observed in sections incubated with biotinylated relaxin (Fig. 4A). No signal was detected in sections incubated with unmodified porcine relaxin (Fig. 4B). Binding of biotinylated relaxin in the mammary glands was hormone-specific and saturable (Fig. 4, C and D).
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Immunohistochemistry results from sections of the mammary nipples are shown in Figure 5. Prominent labeling with the biotinylated relaxin was observed in epithelial cells of lactiferous duct, smooth muscle cells, and skin (Fig. 5A). Prominent labeling was also observed in sebaceous glands (Fig. 5A). Labeling of less intensity was observed in blood vessels (Fig. 5A). No signal was detected in sections incubated with unmodified porcine relaxin (Fig. 5B). Binding of the biotinylated relaxin in the mammary nipples was hormone-specific and saturable (Fig. 5, C and D).
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Immunohistochemistry results from sections of term placenta are shown in Figure 6. Prominent labeling was observed in both amnion in chorionic plate and the large numbers of placental villi projecting into the lacuna system (Fig. 6, A–C). With the higher magnification of the chorionic plate, labeling was observed in amnion epithelial cells and smooth muscle cells surrounding blood vessels (Fig. 6B). Binding was also associated with stromal cells located within the chorionic plate (Fig. 6B). With the higher magnification of the placental villi, prominent labeling was observed in syncytiotrophoblast cells (Fig. 6C). Labeling of less intensity was observed in blood vessels within the villi (Fig. 6C). No signal was detected in sections incubated with unmodified porcine relaxin (Fig. 6D). Binding of the biotinylated relaxin in the term placenta was hormone-specific and saturable (Fig. 6, E and F).
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DISCUSSION |
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TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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Whereas it is possible that biotinylated relaxin binds to other proteins, there are at least three reasons to think that the relaxin-binding sites associated with human cells are functional relaxin receptors. First, relaxin-binding was associated with identical cell types in the cervix, vagina, uterus, mammary glands, and mammary nipples of the human as in rats and pigs [14, 15, 27, 31]. Second, in pigs and rats, relaxin elicits a biological response in all tissues in which cells bind relaxin with specificity. Relaxin induces growth and softening of the cervix [9–12], growth of the vagina and/or uterus [13–15], growth and/or development of the mammary glands [16–18], growth of the mammary nipples [16, 19, 20], expansion of the skin [32], and reduction in the amplitude of spontaneous contractions of the intestine [20]. Third, in this study, specific relaxin binding was found in human skin and placenta, and synthetic human relaxin was recently reported to demonstrate a biological response in human skin and placental tissue [23, 24, 33]. Establishment of receptor binding requires structural characterization of the relaxin receptor and its subsequent localization using specific relaxin-receptor probes. The remainder of this discussion is grounded on the premise that the relaxin-binding sites identified in this study are relaxin receptors.
The physiological significance of relaxin's binding to multiple cell types in human tissues is not yet known. An obvious implication is that relaxin has multiple and complex actions on these tissues. The present findings, as well as the similarity of findings in rats and pigs [14, 15, 27, 31], permit speculation.
Consider first the possible physiological roles of relaxin on epithelial cells, blood vessels, stromal cells (putative fibroblasts), and smooth muscle in the cervix and vagina. Relaxin may have at least two actions on the epithelial cells. Relaxin may enlarge the circumference of the lumen by promoting an increase in epithelial cells as it does in the rat cervix and vagina ([14, 34], unpublished results). Additionally, relaxin may expedite safe delivery by promoting the secretion of mucins [35] into the birth canal. There is limited evidence that relaxin may stimulate increased mucin secretion in the rat cervical epithelium [34]. Relaxin may act directly on the blood vessels to induce their dilation as it does in the rat cervix [34] and vagina (unpublished results). Enlargement of blood vessels may provide an adequate blood supply for increased metabolic activity and/or enhanced migration of polymorphonuclear leukocytes into the cervix during late pregnancy in the human [36]. Polymorphonuclear leukocytes have been postulated to release matrix metalloproteinase enzymes that bring about degradation of the extracellular matrix in the human cervix [36]. Relaxin may act directly on stromal cells to promote remodeling of the extracellular matrix. Relaxin was reported to promote the secretion of several matrix metalloproteinases and to increase the synthesis of glycosaminoglycans in primary cultures of human stromal cells with fibroblast morphology [37]. Endogenous relaxin may act directly on cervical smooth muscle to inhibit its contractility. Norström and coworkers [38] reported that porcine relaxin inhibited spontaneous contractions of strips of human cervix obtained at term pregnancy. Relaxin's quiescent effect on contractility of cervical smooth muscle is of unknown physiological significance because it remains to be demonstrated that reduced contractility of cervical smooth muscle contributes to cervical softening.
Consider second the possible physiological roles of relaxin on epithelial cells, blood vessels, stromal cells, and smooth muscle cells in the uterus. As with the cervix and vagina, relaxin may act directly on uterine epithelial cells to contribute to enlargement of the lumen. Relaxin promotes growth of the pig uterus during pregnancy [15], and Tseng and coworkers [39] reported that porcine relaxin stimulated an increase in cAMP in primary cultures of human endometrial epithelial cells. As with the rat cervix [34] and vagina (unpublished results), relaxin may act directly on blood vessels in the human endometrial stroma to induce their dilation. Relaxin promotes a marked dilation in blood vessels within the mouse endometrial stroma [40]. Relaxin may act on stromal cells within the human endometrium to contribute to their decidualization. When relaxin in combination with progestin was administered to cultured human stromal cells that resembled fibroblasts, there was a dramatic change in the cells to a morphology that resembled decidualized cells [41]. Moreover, the morphological changes were accompanied by an increase in the secretion of prolactin and insulin-like growth factor binding protein-1—two major decidual secretory proteins [41]. Whereas it seems reasonable to speculate that relaxin acts directly on human myometrial smooth muscle to render the uterus quiescent as it does in rats, pigs, mice, guinea pigs, and other species [8], that might not be the case. Both highly purified porcine relaxin [42] and synthetic human relaxin H2 [43, 44] demonstrated little or no effect on spontaneous contractility of human myometrial tissues obtained from either pregnant women at cesarean section or nonpregnant women at hysterectomy.
As postulated for the cervix, vagina, and uterus, relaxin may act directly on both epithelial cells and blood vessels to promote growth and differentiation of the mammary glands and nipples during human pregnancy. Whereas relaxin's effects on growth and/or development of the mammary apparatus are dependent upon circulating luteal relaxin in rats and pigs [16–18, 20], growth and/or development of the mammary glands in the human may be regulated by autocrine and/or paracrine mechanisms. There is limited evidence that human mammary glands produce small amounts of relaxin [22, 45].
The physiological significance of relaxin's binding to the skin of the nipples in the human, rats [31], and pigs [27] is not known. It may cause a remodeling of the extracellular matrix. Unemori and coworkers found that synthetic human relaxin H2 decreased collagen synthesis by human dermal fibroblasts in vitro [46] and decreased collagen accumulation in vivo in two rodent models of fibrosis [47]. Also, s.c. administration of synthetic human relaxin H2 via an osmotic pump over a three-day period was reported to increase the rate of tissue expansion in male piglets [32]. Relaxin may also contribute to the function of the sebaceous glands. Whereas their function is not clearly understood, the sebaceous glands secrete sebum and, in some species, produce pheromones [48].
The identification of relaxin-binding on the amnion epithelial cells, syncytiotrophoblast, blood vessels and stromal cells within the term placenta is consistent with Bryant-Greenwood and coworkers' hypothesis that decidual and/or placental relaxin [21] acts locally via autocrine and/or paracrine mechanisms. The binding of decidual and/or placental relaxin to the amnion epithelial cells and stromal cells may contribute to connective tissue remodeling at the maternal-fetal interface during late pregnancy and at birth [23, 24, 49]. The physiological significance of relaxin binding to syncytiotrophoblasts is more speculative. Relaxin might control the production of proteins such as follistatin [50], transforming growth factor ß1 [51], and interleukin-1 [52], which in turn regulate hCG production. Relaxin may also modulate the transfer of substances between the mother and fetus via autocrine regulation of transport mechanisms such as the glucose transport protein-1, which is localized on the syncytiotrophoblasts of the human placenta and increases as pregnancy progresses [53]. As in the reproductive tract ([34, 40], unpublished results), the binding of relaxin to the placental blood vessels may cause their dilation and thereby contribute to maintenance of adequate blood supply to the fetus.
Our previous studies with pigs [15, 27] and rats [14, 31] demonstrated that endogenous circulating relaxin does not occupy relaxin-binding sites to the extent that it interferes noticeably with binding of the biotinylated relaxin in reproductive and other tissues. In this study, we also found that there were no apparent differences in the distribution and intensity of labeling in cervices obtained from nonpregnant women after hysterectomy and women after cesarean section at term (data not shown). Thus available data indicate that, with the exception of the placenta, the use of tissues from nonpregnant humans are satisfactory for identifying specific relaxin-binding cells.
Findings in this study support the view that relaxin-induced remodeling of connective tissue has potential for clinical applications. The efficacy of human recombinant relaxin for the treatment of systemic sclerosis is presently under investigation [54]. Relaxin also has potential as a cervical softening agent at term. Two recent clinical trials administered recombinant human relaxin intravaginally at term with the apparent intent of exerting a local effect on the cervix while avoiding possible systemic side effects [55, 56]. Both groups of investigators concluded that the failure of the relaxin treatment to soften the cervix may have been attributable to ineffective absorption of relaxin [55, 56]. There is a need to investigate the influence of relaxin on cervical softening in women and other primates under experimental conditions in which the hormone is administered in such a way as to have ready access to the systemic circulation.
In summary, this study identifies specific relaxin-binding cells in the cervix, vagina, uterus, mammary glands, nipples, and placenta of the human. They are epithelial cells, smooth muscle cells, and blood vessels in the cervix, vagina, uterus, and mammary nipples; epithelial cells and blood vessels in the mammary glands; skin and sebaceous glands of the nipples; and amnion epithelium, syncytiotrophoblast, and blood vessels within the placenta. We conclude that the specific relaxin-binding cells probably contain relaxin receptors. Identification of putative relaxin receptors may provide insight into physiological and/or therapeutic roles for relaxin in the human.
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ACKNOWLEDGMENTS |
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FOOTNOTES |
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2 Correspondence: O. David Sherwood, Department of Molecular and Integrative Physiology, University of Illinois at Urbana-Champaign, 524 Burrill Hall, 407 South Goodwin Avenue, Urbana, IL 61801. FAX: 217 333 1133; od-sherw{at}uiuc.edu
Accepted: June 10, 1998.
Received: April 3, 1998.
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REFERENCES |
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TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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-aminohexanoyl) porcine relaxin. Int J Pept Protein Res 1990; 35:416–423.[Medline]This article has been cited by other articles:
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  J. Novak, L. J. Parry, J. E. Matthews, L. J. Kerchner, K. Indovina, K. Hanley-Yanez, K. D. Doty, D. O. Debrah, S. G. Shroff, and K. P. Conrad Evidence for local relaxin ligand-receptor expression and function in arteries FASEB J, November 1, 2006; 20(13): 2352 - 2362. [Abstract] [Full Text] [PDF]
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 W. Yan, A. A. Wiley, R. A. D. Bathgate, A.-L. Frankshun, S. Lasano, B. D. Crean, B. G. Steinetz, C. A. Bagnell, and F. F. Bartol Expression of LGR7 and LGR8 by Neonatal Porcine Uterine Tissues and Transmission of Milk-Borne Relaxin into the Neonatal Circulation by Suckling Endocrinology, September 1, 2006; 147(9): 4303 - 4310. [Abstract] [Full Text] [PDF]
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 R. A. Bathgate, R. Ivell, B. M. Sanborn, O. D. Sherwood, and R. J. Summers International Union of Pharmacology LVII: Recommendations for the Nomenclature of Receptors for Relaxin Family Peptides. Pharmacol. Rev., March 1, 2006; 58(1): 7 - 31. [Abstract] [Full Text] [PDF]
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 E.S. Hayes, E.C. Curnow, A.O. Trounson, L.A. Danielson, and E.N. Unemori Implantation and Pregnancy Following In Vitro Fertilization and the Effect of Recombinant Human Relaxin Administration in Macaca fascicularis Biol Reprod, November 1, 2004; 71(5): 1591 - 1597. [Abstract] [Full Text] [PDF]
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 K. P. Conrad and J. Novak Emerging role of relaxin in renal and cardiovascular function Am J Physiol Regulatory Integrative Comp Physiol, August 1, 2004; 287(2): R250 - R261. [Abstract] [Full Text] [PDF]
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 C. P. Bond, L. J. Parry, C. S. Samuel, H. M. Gehring, F. L. Lederman, P. A. W. Rogers, and R. J. Summers Increased Expression of the Relaxin Receptor (LGR7) in Human Endometrium during the Secretory Phase of the Menstrual Cycle J. Clin. Endocrinol. Metab., July 1, 2004; 89(7): 3477 - 3485. [Abstract] [Full Text] [PDF]
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 S. Quattrone, L. Chiappini, G. Scapagnini, B. Bigazzi, and D. Bani Relaxin potentiates the expression of inducible nitric oxide synthase by endothelial cells from human umbilical vein in in vitro culture Mol. Hum. Reprod., May 1, 2004; 10(5): 325 - 330. [Abstract] [Full Text] [PDF]
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 O. D. Sherwood Relaxin's Physiological Roles and Other Diverse Actions Endocr. Rev., April 1, 2004; 25(2): 205 - 234. [Abstract] [Full Text] [PDF]
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 M. A. M. Krajnc-Franken, A. J. M. van Disseldorp, J. E. Koenders, S. Mosselman, M. van Duin, and J. A. Gossen Impaired Nipple Development and Parturition in LGR7 Knockout Mice Mol. Cell. Biol., January 15, 2004; 24(2): 687 - 696. [Abstract] [Full Text] [PDF]
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 J. L. Dragoo, R. S. Lee, P. Benhaim, G. A. M. Finerman, and S. L. Hame Relaxin Receptors in the Human Female Anterior Cruciate Ligament Am. J. Sports Med., July 1, 2003; 31(4): 577 - 584. [Abstract] [Full Text] [PDF]
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A. Einspanier, D. Muller, J. Lubberstedt, O. Bartsch, A. Jurdzinski, K. Fuhrmann, and R. Ivell Characterization of relaxin binding in the uterus of the marmoset monkey Mol. Hum. Reprod., October 1, 2001; 7(10): 963 - 970. [Abstract] [Full Text] [PDF]
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O. Bartsch, B. Bartlick, and R. Ivell Relaxin signalling links tyrosine phosphorylation to phosphodiesterase and adenylyl cyclase activity Mol. Hum. Reprod., September 1, 2001; 7(9): 799 - 809. [Abstract] [Full Text] [PDF]
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L. A. Danielson, L. J. Kercher, and K. P. Conrad Impact of gender and endothelin on renal vasodilation and hyperfiltration induced by relaxin in conscious rats Am J Physiol Regulatory Integrative Comp Physiol, October 1, 2000; 279(4): R1298 - R1304. [Abstract] [Full Text] [PDF]
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 S. Y. Hsu Cloning of Two Novel Mammalian Paralogs of Relaxin/Insulin Family Proteins and Their Expression in Testis and Kidney Mol. Endocrinol., December 1, 1999; 13(12): 2163 - 2174. [Abstract] [Full Text]
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