An enzyme-linked immunosorbent assay to study human relaxin in human pregnancy and in pregnant rhesus monkeys

Lucas, Cathy H.; Bald, Laura; Martín, María Cecilia; Jaffe, Richard; Drolet, Daniel; Mora-Worms, Marina; Bennett, Greer A.; Chen, A. B.; Johnston, Paul D.

doi:10.1677/joe.0.1200449

Cited by 71 publications

(30 citation statements)

References 27 publications

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“…The sensitivity of the Rlx RIA was 31-125 pg porcine Rlx equivalents per tube, as determined in individual assays. (As has been our experience with serum Rlxs from other monkeys, the marmoset serum Rlx did not give a detectable reading in the homologous human Rlx ELISA developed by Lucas et al [36] and generously provided by Richard Vandlen of Genentech Inc., So. San Francisco, CA).…”

Section: Methodsmentioning

confidence: 94%

Patterns of Relaxin and Steroids in the Reproductive Cycle of the Common Marmoset (Callithrix Jacchus): Effects of Prostaglandin F2α on Relaxin and Progesterone Secretion during Pregnancy

Steinetz¹,

Randolph²,

Mahoney³

1995

Biology of Reproduction

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We measured the concentrations of relaxin (Rlx), progesterone, and estradiol-17 beta in serum samples obtained twice or three times weekly from marmosets during the estrous cycle and pregnancy. The cyclic patterns and concentrations of progesterone and estradiol-17 beta were similar to those reported by previous investigators. Rlx was not detected in individual serum samples ( < 0.62-1.25 ng/ml) obtained from nonpregnant marmosets. However, pooling of luteal serum from all animals permitted assay of much larger volumes of serum (0.4 ml vs. 0.1 ml), and a concentration of about 1 ng/ml was detected. Rlx was first detected in serum in the second or third week of the 21-wk marmoset pregnancy, rose to a peak during Weeks 10-14, and then declined slowly as the time of parturition approached. The pattern of Rlx was unlike that observed during pregnancy in Old World monkeys, chimpanzees, or women, and resembled, instead, that seen in rodents, carnivores, and equids. Progesterone and estradiol-17 beta likewise increased throughout pregnancy, and their patterns were similar to those previously described for marmosets by other investigators. The concentrations of the steroids and Rlx in serum of pregnant marmosets was 10-fold or more higher than those found in Old World monkeys, baboons, chimpanzees, or women. Spontaneous abortions in two of the marmosets were accompanied by precipitous falls in serum levels of progesterone, estradiol-17 beta, and Rlx. Following s.c. injection of the luteolytic agent prostaglandin F2 alpha (PGF2 alpha) into two marmosets at midpregnancy, serum progesterone and Rlx fell to low levels. These animals received a progestin, 17 alpha-ethyl-19-nortesterone, to preclude abortion. Serum progesterone rose again, but serum Rlx remained low for the duration of pregnancy.(ABSTRACT TRUNCATED AT 250 WORDS)

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Section: Methodsmentioning

confidence: 94%

Patterns of Relaxin and Steroids in the Reproductive Cycle of the Common Marmoset (Callithrix Jacchus): Effects of Prostaglandin F2α on Relaxin and Progesterone Secretion during Pregnancy

Steinetz¹,

Randolph²,

Mahoney³

1995

Biology of Reproduction

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“…Many studies have reported circulating levels of relaxin in human subjects using an ELISA developed by Genentech (26). We have tested the ability of the anti-H2 relaxin antibody used in this ELISA to recognize H1 and H3 relaxins.…”

Section: Discussionmentioning

confidence: 99%

Human Relaxin Gene 3 (H3) and the Equivalent Mouse Relaxin (M3) Gene

Bathgate

Samuel

Burazin

et al. 2002

Journal of Biological Chemistry

360

396

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We have identified a novel human relaxin gene, designated H3 relaxin, and an equivalent relaxin gene in the mouse from the Celera Genomics data base. Both genes encode a putative prohormone sequence incorporating the classic two-chain, three cysteine-bonded structure of the relaxin/insulin family and, importantly, contain the RXXXRXX(I/V) motif in the B-chain that is essential for relaxin receptor binding. A peptide derived from the likely proteolytic processing of the H3 relaxin prohormone sequence was synthesized and found to possess relaxin activity in bioassays utilizing the human monocytic cell line, THP-1, that expresses the relaxin receptor. The expression of this novel relaxin gene was studied in mouse tissues using RT-PCR, where transcripts were identified with a pattern of expression distinct from that of the previously characterized mouse relaxin. The highest levels of expression were found in the brain, whereas significant expression was also observed in the spleen, thymus, lung, and ovary. Northern blotting demonstrated an ϳ1.2-kb transcript present in mouse brain poly(A) RNA but not in other tissues. These data, together with the localization of transcripts in the pars ventromedialis of the dorsal tegmental nucleus of C57BLK6J mouse brain by in situ hybridization histochemistry, suggest a new role for relaxin in neuropeptide signaling processes. Together, these studies describe a third member of the human relaxin family and its equivalent in the mouse.Relaxin is a 6-kDa polypeptide hormone that is secreted by the ovary into the peripheral circulation in highest amounts during pregnancy and has a number of functions in mammals that are generally associated with female reproductive tract physiology (1). To date, only one relaxin gene has been characterized in most mammalian species, with the exception of the human where two separate genes have been described, designated H1 (2) and H2 (3) relaxin. The peptide encoded by the H2 gene is the major stored and circulating form in the human (4). H1 relaxin expression is restricted to the decidua, placenta, and prostate (5); however, the H1 peptide has similar biological activity to that of H2 relaxin in a rat atrial bioassay (6). The actions of relaxin include an ability to inhibit myometrial contractions, to stimulate remodeling of the connective tissue, and to induce softening of the tissues of the birth canal. Additionally, relaxin increases growth and differentiation of the mammary gland and nipple and induces the breakdown of collagen, one of the main components of connective tissue. Relaxin decreases collagen synthesis and increases the release of collagenases (7). These findings were recently confirmed by the establishment of the relaxin gene-knockout mouse (8), which exhibited a number of phenotypic properties associated with pregnancy. Female mice lacking a functionally active relaxin gene failed to relax and elongate the interpubic ligament of the pubic symphysis and could not suckle their pups, who in turn died within 24 h unless cross-fostered t...

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“…Relaxin concentrations were measured by an ELISA (Genentech Inc, Calif.) based on purified antibody raised against a recombinant human relaxin (hRLX-2) as described previously [6], and the results were calculated from the standard curves which were correlated with a known standard concentration measured in all assays. All samples were tested in one assay.…”

Section: Methodsmentioning

confidence: 99%

Normal Serum Relaxin in Women with Disabling Pelvic Pain during Pregnancy

Petersen

Hvidman

Uldbjerg

1994

Gynecol Obstet Invest

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As part of a case control within cohort study 472 pregnant women answered a questionnaire post partum. Serum relaxin concentrations were measured by a homologous ELISA in samples collected from the women in the 30th week of pregnancy. Serum relaxin concentrations were not associated with pregnancy-associated pelvic pain. Thus, normal pregnant women without pelvic pain (n = 118) had mean concentrations of 343 pg/ml compared to 332 pg/ml in women (n = 59) with pelvic pain and some restriction in daily activities and to 349 pg/ ml in women (n = 9) with severe pregnancy-induced pelvic pain. The present results do not suggest a role for relaxin in symptom-giving pelvic relaxation during human pregnancy.

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An enzyme-linked immunosorbent assay to study human relaxin in human pregnancy and in pregnant rhesus monkeys

Cited by 71 publications

References 27 publications

Patterns of Relaxin and Steroids in the Reproductive Cycle of the Common Marmoset (Callithrix Jacchus): Effects of Prostaglandin F2α on Relaxin and Progesterone Secretion during Pregnancy

Patterns of Relaxin and Steroids in the Reproductive Cycle of the Common Marmoset (Callithrix Jacchus): Effects of Prostaglandin F2α on Relaxin and Progesterone Secretion during Pregnancy

Human Relaxin Gene 3 (H3) and the Equivalent Mouse Relaxin (M3) Gene

Normal Serum Relaxin in Women with Disabling Pelvic Pain during Pregnancy

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