Relaxin in Ovarian Tissue During Different Reproductive Stages in the Rat

Anderson, L. L.; Bast, Joseph D.; Melampy, R. M.

doi:10.1677/joe.0.0590371

Cited by 48 publications

(12 citation statements)

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“…Anderson et al (7) reported a similar increase in ovarian relaxin (bioassay) between Days 14 and 20 of pregnancy in rats. The levels rose between the twelfth and fifteenth day of gestation and remained elevated until after parturition.…”

Section: Research Department Pharmaceutical Division Ciba -Geigy Comentioning

confidence: 74%

Radioimmunoassay (RIA) of Relaxin in Sera of Various Species Using an Antiserum to Porcine Relaxin

O’Byrne¹,

Steinetz²

1976

Experimental Biology and Medicine

154

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Recent work from our laboratory led to the development of a radioimmunoassay (RIA) for porcine relaxin utilizing [ 12.51]polytyrosyl relaxin (1 ) . Addition of tyrosine to the relaxin molecule proved to be necessary as the purified native hormone does not contain tyrosine or histidine (2) and fails to iodinate when subjected to the Hunter and Greenwood method (1). An impure relaxin extract was used by other workers who claimed successful iodination of the hormone (3).We have previously demonstrated crossreactions of antisera to porcine relaxin with relaxin-con taining tissues of several species including rats, mice, chickens, whales, and human beings (4). Because of this broad range of immunological cross-reactivity of relaxin from different sources, it was of interest to develop a RIA for determination of blood levels of relaxin in species other than the pig. We have now assayed relaxin levels throughout pregnancy in rats, mice, and guinea pigs, and near term in dogs and primates.Materials and methods. Radioimmunoassay. The relaxin RIA has been modified since the publication of the original report Reagents. Two fractions of porcine relaxin prepared according to the procedure of Sherwood and O'Byrne (2) were used in the development of the method. A moderately pure fraction (G-50:1000 U/mg) was used for immunization and a highly purified fraction (CM-A + B: 2500 to 3000 U/mg) was used for standards and preparation of polytyrosyl-relaxin . The assay was equally sensitive and linear when we used a purified relaxin prepared by a different method and kindly supplied by Dr . Christian Schwabe, Medical College of South Carolina. Thus, the antigenic determinants of porcine relaxin do not appear to be altered by differing routes of biochemical purification. How-(1). ever, reduced-alkylated relaxin as well as isolated a and p subunits of relaxin did not compete with [1251]polytyrosyl-relaxin for binding sites on the antibody. Although relaxin precursors and/or metabolic products may be immunoreactive, these observations suggest a requirement for the overall structure of two peptide chains connected by disulfide bonds for antibody recognition. Polytyrosyl-relaxin ,[ 1251]polytyrosyl-relaxin, and relaxin antisera were prepared as previously described (1). Goat anti-rabbit gamma globulin was purchased from Antibodies Incorporated, Davis, California.RIA procedure. 1. One-hundred microliters of relaxin standard solutions [ l o to 2500 pg relaxin in phosphate buffered saline (PBS = 0.01 M sodium phosphate, pH 7.0, 0.14 M sodium chloride)-1% egg albumin] plus a volume of control serum or plasma (male or castrated) equivalent to that of the unknown samples were added to standard curve tubes.2. Unknown serum or plasma samples were added to other tubes.3 . Sufficient PBS-1% egg albumin was added to each tube to bring the volume to 500 pl. 4. One-hundred microliters of [12"I]polytyrosyl-relaxin in PBS-1% egg albumin (30,000 to 60,000 cpm) were added to each tube.5 . One-hundred microliters of relaxin antiserum diluted in 0.05 M EDTA-...

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Section: Research Department Pharmaceutical Division Ciba -Geigy Comentioning

confidence: 74%

Radioimmunoassay (RIA) of Relaxin in Sera of Various Species Using an Antiserum to Porcine Relaxin

O’Byrne¹,

Steinetz²

1976

Experimental Biology and Medicine

154

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“…The increase in cervical dilatability which oc curs during pregnancy in rats is correlated with the first appearance and subsequent rise in blood levels of immunoreactive relaxin 5,10 and with in creasing ovarian concentrations of relaxin mea sured both by RIA 11,12 and by bioassay. 13 Removal of the ovaries results in a rapid loss of measurable serum immunoreactive relaxin; 10,14 and, when ovariectomy is performed on day 15 of pregnancy, the cervix fails to soften. 5,15,16 This evidence, together with relaxin bioassays of var ious tissues (see Sherwood et al 10 for review), suggests that the ovaries are the major, if not the only source of circulating relaxin in the rat.…”

Section: Cervical Changes During Pregnancy In Ratsmentioning

confidence: 99%

“…20 Relaxin also stim ulates release of prostaglandins 22 in a pushpull type situation at this time and also facilitates the action of PGs and oxytocin on the uterus. 13,36 Uterine contractions become coordinated due to the withdrawal of progesterone and the relaxin enhanced response of the myometrium to oxy tocic substances. Parturition occurs, and the abrupt withdrawal of progesterone, prostaglan dins and relaxin, together with the increasing, unopposed activity of estrogens results in the rapid loss of cervical water and regression of the collagen framework back to the rigid pre-relaxin state, before involution of the cervical mass is completed.…”

Section: Cervical Changes During Pregnancy In Ratsmentioning

confidence: 99%

Speculations on the Probable Role of Relaxin in Cervical Dilation and Parturition in Rats

Steinetz¹,

O’Byrne²

1983

Semin Reprod Med

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“…The corpora lutea (CL) are the source of the polypeptide hormone relaxin in the pregnant rat (Anderson & Long, 1978;Hudson, Haley, Cronk, Shine & Niall, 1981;Golos, Weyhenmeyer & Sherwood, 1984). Relaxin concentrations increase in the ovaries and serum from around Day 10 of pregnancy until Day 20 and decline rapidly 1 or 2 days before birth (Anderson, Bast & Melampy, 1973;Sherwood, Crnekovic, Gordon & Rutherford, 1980). The placenta is required for the promotion and maintenance of high concentrations of relaxin in the serum and ovaries (Goldsmith et ai, 1981 ; Golos & Sherwood, 1982), as it is for CL growth and progesterone secretion (Takayama & Greenwald, 1973;Gibori, Antczak & Rothchild, 1977; Kato, Morishige & Rothchild, 1979; Golos & Sherwood, 1982).…”

Section: Introductionmentioning

confidence: 99%

Effect of oestradiol-17 on ovarian and serum concentrations of relaxin during the second half of pregnancy in the rat

Guico¹,

Sherwood²

1985

Reproduction

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Immunoactivity concentrations of ovarian relaxin, serum relaxin and serum progesterone were determined from Day 12 through Day 18 of pregnancy in rats treated with oil or oestradiol-17\g=b\ after hysterectomy or hypophysectomy and hysterectomy on Day 12. Relaxin and progesterone concentrations increased between Days 12 and 18 in sham-operated rats but failed to increase or declined in oil-treated hysterectomized or hypophysectomized\p=n-\hysterectomized animals. Oestradiol treatment increased serum concentrations of relaxin and progesterone in hypophysecto-mized\p=n-\hysterectomized rats on Day 15 and increased the concentrations of ovarian and serum relaxin and serum progesterone in hysterectomized rats on Day 18. These data are consistent with the concept that placental support for the promotion and maintenance of relaxin and progesterone concentrations from Day 12 through Day 18 may be mediated, at least in part, through a common mechanism(s) which involves oestradiol.

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Relaxin in Ovarian Tissue During Different Reproductive Stages in the Rat

Cited by 48 publications

References 0 publications

Radioimmunoassay (RIA) of Relaxin in Sera of Various Species Using an Antiserum to Porcine Relaxin

Radioimmunoassay (RIA) of Relaxin in Sera of Various Species Using an Antiserum to Porcine Relaxin

Speculations on the Probable Role of Relaxin in Cervical Dilation and Parturition in Rats

Effect of oestradiol-17 on ovarian and serum concentrations of relaxin during the second half of pregnancy in the rat

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