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The transforming growth factor alpha (TGF-alpha) and the epidermal growth factor (EGF) control the production of mouse placental lactogen I (mPL-I) and mPL-II in different manners. EGF enhances the action of mPL-I, while TGF-alpha inhibits the secretion of the hormone. Their actions on differentiation of the mPL-I and mPL-II producing cells are also different. However, TGF-alpha and EGF exhibit around 40 percent similarity in the amino acid sequence and act by binding to a common receptor.

The purpose of this study was to investigate whether the ballistic push-up (BPU) is responsive to post-activation performance enhancement (PAPE) after a bench press conditioning exercise using velocity-based repetition control. Additionally, we aimed to evaluate the effects of range of motion (ROM) conditions on subsequent BPU performance. In a randomized crossover design, 18 males performed two conditions (full ROM and self-selected partial ROM) of bench press at 80% of their 1RM until mean concentric velocity dropped 10%. Each participant performed two pre- and six post-test BPUs to assess the PAPE effect. Paired sample t-tests assessed bench press performance measures. Multiple two-way repeated measures ANOVAs assessed differences in flight time, impulse, and peak power for the pre- and post-test BPUs. No significant differences existed between ROM conditions for total repetitions, volume load, or peak velocity. Compared to partial ROM, full ROM showed greater displacement (0.42 ± 0.05 vs. 0.34 ± 0.05 m), work (331.99 ± 67.72 vs. 270.92 ± 61.42 J), and mean velocity (0.46 ± 0.09 vs. 0.44 ± 0.08 m/s). Neither bench press ROM condition enhanced the BPU and were detrimental in some cases. Several time points showed partial ROM (flight time: 2 min post, impulse: 12 min post, peak power: 12 min post) significantly greater than full ROM, possibly indicating less fatigue accumulation. The BPU may require a different stimulus or may not be practical for PAPE effects in college-aged males. Partial ROM can be an alternative that achieves similar peak velocities while requiring less overall work.

The placental members of the prolactin-GH-placental lactogen (PL) gene family of the mouse include mPL-I, mPL-II, proliferin (PLF) and proliferin-related protein (PRP). The aim of the present study was to assess the effects of tumour necrosis factor-alpha (TNF-alpha) on the secretion of these proteins in primary cultures of placental cells from days 7, 9 and 12 of pregnancy. The effects of epidermal growth factor (EGF) on the secretion of PLF and PRP were also determined. EGF has previously been shown to stimulate mPL-I and inhibit mPL-II secretion. Incubation of placental cells from day 7 of pregnancy for 5 days with 10 nmol human (h)TNF-alpha/l did not affect the mPL-II concentration of the medium, but similar treatment of cells from days 9 or 12 of pregnancy resulted in a significant reduction in the mPL-II concentration of the medium by the second or third day of culture. The intracellular concentration of mPL-II, the number of cells that released mPL-II as assessed by reverse haemolytic plaque assay, and steady-state levels of mPL-II mRNA as assessed by Northern analysis were also reduced by hTNF-alpha treatment. The lowest concentration of hTNF-alpha that significantly inhibited mPL-II secretion by cells from day 12 of pregnancy was 0.01 nmol/l. hTNF-alpha treatment did not affect the secretion of mPL-I, PLF or PRP, as assessed by the concentrations of these proteins in the medium during a 5-day incubation. Incubation of the cells with 20 ng EGF/ml also did not affect the PLF or PRP concentration of the medium during 5 days of culture. To determine whether the effect of hTNF-alpha on mPL-II secretion was mediated by interleukin-6 (IL-6), the IL-6 concentration of the medium of control and hTNF-alpha-treated cells was determined. Bioactive and immunoreactive IL-6 could not be detected in medium from either treatment group. The presence of binding sites for hTNF-alpha was assessed in cells from day 12 of pregnancy. Scatchard analysis detected a single class of binding sites having a Kd of 1.61 +/- 0.34 nmol/l, with about 1350 sites per cell. The results of this study demonstrate that hTNF-alpha inhibits the secretion of mPL-II by placental cells from days 9 and 12 of pregnancy, suggesting that TNF-alpha may be one of the factors that regulate the production of this hormone in vivo.

Swiss Webster mice were hypophysectomized or sham-operated on Day 11 of pregnancy. The animals were fitted s.c. with osmotic minipumps containing either corticosterone (B) dissolved in Molecusol (Pharmatec, Alachua, FL) or the vehicle alone immediately after they were hypophysectomized. Animals in some of the experimental groups also received thyroxine (T4) in their drinking water. The mice were killed on Day 18 of gestation, and mammary tissue was homogenized and extracted for assessment of DNA, RNA, alpha-lactalbumin, and alpha-casein. Serum was assayed for placental lactogen-I (PL-I), and placental lactogen-II (PL-II), B, and T4. The concentration of PL-II in serum was elevated in the hypophysectomized mice, whereas the PL-I concentration did not differ among experimental groups. Hypophysectomy decreased both T4 and B concentrations in serum, and administration of these hormones restored their serum concentrations to normal or, in some cases, somewhat higher than normal levels. Hypophysectomy reduced the total RNA content and RNA/DNA ratio of the mammary gland, but treatment with B alone or with B and T4 restored RNA levels to those of sham-operated animals. T4 alone was ineffective in restoring RNA levels. Sham-operated animals that received hormonal treatment (B and T4) had the highest levels of RNA in the mammary tissue. Hypophysectomized animals had reduced content and concentration of alpha-lactalbumin in the mammary gland as compared to all other experimental groups.(ABSTRACT TRUNCATED AT 250 WORDS)

The purpose of this investigation was to identify the cellular origin of placental lactogen-I (PL-I) expression in the mouse placenta and to cytologically define the transition from PL-I to PL-II expression during gestation. PL-I mRNA expression was assessed by in situ hybridization, and expression of PL-I and PL-II protein was determined by immunocytochemical analysis. PL-I mRNA and protein were localized to trophoblast giant cells. Trophoblast giant cells ceased producing PL-I at midgestation and began expressing PL-II. PL-I immunoreactivity was present in trophoblast giant cells on Days 9 and 10 of gestation but was not detectable in trophoblast giant cells on Day 11 of gestation. Immunoreactive PL-II-producing giant cells were detected first on Day 10 of gestation, continuing on Day 11 of gestation. Expression of PL-I and PL-II signals a significant functional transition in trophoblast giant cells of the developing mouse placenta.

The occurrence of placental lactogens has been reported for members of 4 orders, 7 families, and 15 genera, while its absence has been documented in representatives from 4 orders, 5 families, and 5 genera. Placental lactogens from several genera have been isolated, purified, and characterized biochemically. The cellular sites of production and patterns of secretion of placental lactogens have been described. The regulation of placental lactogen secretion is not understood, although its synthesis and release can be modified experimentally by altering metabolic, ionic, and hormonal factors in the placental environment. Placental lactogens stimulate lactogenesis and steroidogenesis. The mechanism underlying placental lactogen's role in these processes has not been clarified.

Changes in nesting frequency, serum LH, progesterone, and estradiol were measured in relation to the onset of broodiness in turkeys. In each of two experiments the frequency of nest visits increased about four-fold in the 5 to 7 day interval before oviposition of the last egg associated with the onset of broodiness. In the 9 day period prior to oviposition of this egg LH levels declined from about 12 ng/ml to 6 to 7 ng/ml (Experiment 1) and from about 9 ng/ml to less than 5 ng/ml (Experiment 2). Progesterone levels declined during this same period going from about 2 to 2.5 ng/ml down to less than .5 ng/ml. In the 5 day period following the nadir in serum LH, which occurred on or a day or two after the last egg, levels of this hormone again rose substantially. Estradiol levels, assayed in Experiment 2, were generally low with many samples below detectability (less than 3 pg/ml). There were no consistent changes in serum estradiol until the day before oviposition of the last egg. On this day, and on the subsequent four days, mean estradiol levels were at a nadir (6 to 9 pg/ml) and rose to levels averaging over 11 pg/ml in the following five day period.

Two PRL-like glycoprotein hormone complexes were purified from the medium of cultured mouse conceptuses from day 10 of pregnancy: mouse placental lactogen-I (mPL-I) (29-32K), and mPL-I (36.5-42K). Sodium dodecyl sulfate-gel electrophoresis revealed that mPL-I (36.5-42K) is a complex of five proteins with mol wt of 36.5K, 37.5K, 39K, 40.5K, and 42K. Deglycosylation with peptide: N-glycosidase F or trifluoromethanesulfonic acid produced a single 29K protein. mPL-I (36.5-42K) was also sensitive to neuraminidase, but not to endo-beta-N-acetylglucosaminidase H or bacterial alkaline phosphatase. The production of intermediates from partial digestion of mPL-I (36.5-42K) with endo-beta-N-acetylglucosaminidase F indicated the presence of multiple glycosylation sites. mPL-I (29-32K) is a complex of three proteins with mol wt of 29K, 30.5K, and 32K. Treatment with peptide:N-glycosidase F or trifluoromethanesulfonic acid reduced the mol wt of the 30.5K and 32K bands to 28K. The 30.5K band was sensitive to endo-beta-N-acetylglucosaminidase H and endo-beta-N-acetylglucosaminidase F, but the 32K band was not. Neither band was sensitive to neuraminidase or bacterial alkaline phosphatase. The 29K band was resistant to all chemical and enzymatic treatments and is probably not glycosylated or phosphorylated. In the nonreduced state, neither form of mPL-I showed an increase in mobility over that of its reduced counterpart on sodium dodecyl sulfate-gel electrophoresis, indicating that neither form of mPL-I contains the large disulfide loop common to hormones of the PRL family. After iodination, all component proteins of both forms of mPL-I were found to bind to day 17 pregnant mouse liver membranes and were displaceable by excess mPL-II. In a radioreceptor assay, 125I-labeled mPL-I (36.5-42K) was displaced by mPRL or mPL-II, but not by mGH. An antiserum to both forms of mPL-I was generated, and a RIA employing mPL-I (36.5-42K) as the standard and radioligand was developed. Dilutions of day 10 pregnant maternal mouse serum and placental homogenate and a partially purified fraction of mPL-I (29-32K) produced displacement curves parallel to that of mPL-I (36.5-42K) standard curve. Five micrograms of mPRL, mPL-II, or mGH or 10 microliter day 17 pregnant or male mouse serum did not displace the radioligand from the antibody. mPL-I (36.5-42K) was lactogenic, but it did not possess LH-like bioactivity.

The effects of perinatal exposure to synthetic estrogens and progestins on mammary tumorigenesis were studied in female C3H/HeN/MTV + mice. Mice were treated neonatally with 0.001 microgram/day diethylstilbestrol (DES), with 15 micrograms/day 17 alpha-hydroxyprogesterone caproate (HPC), or with oil on days 1-5 of life (birth = day 1). As adults, neonatally hormone-treated mice received long-term treatment with a synthetic estrogen and progestin combination or vehicle. Animals were palpated weekly for mammary gland tumors. The effect of treatment on the probability of tumor development was examined. Neonatal treatment with a low dose of DES increased the probability of mammary-gland tumor formation, whereas neonatal treatment with HPC had a slightly protective effect on tumorigenesis. Subsequent treatment of adult mice with synthetic steroids did not affect mammary gland tumorigenesis in neonatally DES-treated or oil-treated animals. There was a significant interaction between the effect of neonatal HPC treatment and subsequent steroid treatment on mammary tumorigenesis but examination of the data indicated that this interaction was due to the protective effect of HPC in the absence of subsequent exposure to synthetic steroids and the probability of tumor appearance in mice treated with both HPC and synthetic steroids as adults did not differ from that of neonatally oil-treated controls.

The effects of neonatal diethylstilbestrol (DES) treatment on pituitary PRL response to dopamine (DA) inhibition or estrogen stimulation were studied in C3H/MTV+ mice at 10 weeks, 5 months and 8 months of age using synthesis and release of [3H]PRL by the pituitary as endpoints. In the DA study, anterior pituitaries from ovariectomized or intact, neonatally hormone-treated mice were incubated for 6 h in the presence or absence of DA. In the estrogen study, serum and anterior pituitaries were obtained from ovariectomized neonatally hormone-treated mice injected with oil or 17 beta-estradiol for 7 days before death. Neonatal treatment of mice with 2.5 micrograms/day DES altered basal [3H]PRL synthesis and release as well as circulating PRL concentrations. The results of analysis of variance indicated a significant interaction between the effects of neonatal hormone treatment and DA treatment on [3H]PRL release. Neonatal DES treatment did not alter the response of pituitary PRL to estrogen treatment at the ages examined.

Hypophysectomy of mice during the last half of pregnancy results in an increase in the maternal serum mouse placental lactogen-II (mPL-II) concentration. The present study was undertaken to isolate and identify the pituitary factors that regulate mPL-II. Administration of pituitary extracts from virgin and day 12 pregnant mice (five pituitaries per mouse, every 8 h, sc) and bilateral pituitary implants under the kidney capsule significantly inhibited the increase in the mPL-II concentration that occurs within 24 h after hypophysectomy on day 12 of pregnancy, which indicated that the pituitary secretes factors that regulate mPL-II. To characterize these factors biochemically, pituitaries from virgin female mice were incubated for 14 days, and the culture medium was pooled, concentrated, and then fractionated by gel exclusion chromatography on Sephadex G-100, followed by ion exchange chromatography on DEAE. The fraction from DEAE chromatography that contained predominantly mouse (m) GH as well as a very small amount of a substance with an approximate mol wt of 23K suppressed the posthypophysectomy rise in serum mPL-II. To determine whether purified PRL, GH, and the unknown 23K substance present in the active fraction from the DEAE column could suppress the posthypophysectomy rise in serum mPL-II, repeated injections and constant infusion of these substances were carried out in hypophysectomized mice. Repeated injections of purified mPRL and bovine (b) PRL and constant infusion of mPRL were without effect. Repeated injection of mGH at doses of 1 and 10 micrograms/mouse or of bGH at doses of 50, 150, and 450 micrograms/mouse were also not effective, but bGH injections at a dose of 5 mg/mouse suppressed the posthypophysectomy rise in serum mPL-II. Subsequently, two pools of mGH were prepared by ion exchange HPLC; mGH-A contained predominantly mGH as well as a very small amount of an unidentified 23K mol wt substance, and mGH-B contained only mGH. When mGH-A and mGH-B were infused at a constant rate into hypophysectomized pregnant mice, both suppressed serum mPL-II concentrations; there was no difference in activity between mGH-A and mGH-B. These results indicate that mGH, but not mPRL, regulates the serum PL-II concentration in the mouse. mPL-II secretion is under the inhibitory control of mGH, the concentration of which increases rapidly at the beginning of the last half of pregnancy.

Previously, we demonstrated that measurement of serum lactogenic hormones during pregnancy in the mouse by the rabbit mammary gland radioreceptor assay (RRA) and specific RIAs for mouse PRL (mPRL) and mouse placental lactogen (mPL) showed a large RRA to RIA activity ratio during midpregnancy. The purpose of this study was to begin to characterize this discrepancy in midpregnancy PRL-like activity. The elution profile of serum from midpregnant mice on Sephadex G-100 showed a characteristic peak in RRA activity (elution volume/void volume = 1.5), distinct from either mPRL or mPL (both approximately elution volume/void volume = 2.0). This PRL-like activity was not recognized by antisera to either mPRL or mPL. The source of the midpregnancy lactogen was localized to the conceptus. The hormone was found to have an apparent molecular weight of 50,000 (based on Sephadex G-100 chromatography) and an electrophoretic mobility similar to mPRL (Rf = 0.5-0.6) and to be lactogenic in the in vitro mouse mammary gland explant bioassay. We have demonstrated the existence of a midpregnancy lactogen that is different from mPL and mPRL and is of conceptus origin.

Swiss Webster mice were hypophysectomized on Day 10 of pregnancy and the effect of the ablation on mammary gland development was estimated by measuring the total weight and the DNA, RNA, and alpha-lactalbumin contents and concentrations of the mammary gland on Days 14 and 18 of gestation. Although a significant increase in mammary gland weight occurred in the hypophysectomized animals between Days 10 and 18 of pregnancy, the mammary gland weight of the hypophysectomized mice was significantly reduced when compared with intact and sham-operated mice on both Days 14 and 18 of pregnancy. The total RNA content of the mammary gland was also reduced in the hypophysectomized mice, although it increased significantly from Day 10 to Day 18. The alpha-lactalbumin content of the mammary gland increased only slightly between Days 10 and 14 of gestation in the intact and sham-operated mice, but a large increase was found on Day 18 in both groups. There was, on the other hand, no increment in the alpha-lactalbumin content of the mammary gland in the hypophysectomized mice either on Day 14 or 18 of gestation. The DNA content of the mammary gland was not affected by hypophysectomy when estimated on Days 14 and 18 of pregnancy. The effects of hypophysectomy on the concentrations of mouse placental lactogen II (mPL-II), progesterone, corticosterone, and thyroxine in the maternal serum were also assessed. The concentration of mPL-II was significantly elevated in the hypophysectomized mice, whereas the circulating concentrations of both corticosterone and thyroxine were greatly reduced. The serum progesterone concentration was not significantly altered by hypophysectomy. (ABSTRACT TRUNCATED AT 250 WORDS)

Pituitary responsiveness to dopamine was investigated on several days of pregnancy in the mouse. Sera and anterior pituitary glands were obtained at 09.00 and 18.00 h on day 5 of pregnancy and at 09.00 h on days 12 and 18, and the pituitaries were incubated for 5 h in several concentrations of dopamine (0, 5 X 10(-10)--5 X 10(-7)M). Serum prolactin (Prl) concentration was the highest on day 5 (18.00 h sample), followed by day 18 (09.00 h), day 5 (09.00 h) and day 12 (09.00 h). Pituitary responsiveness to dopamine was assessed on each day of pregnancy by determining the slopes of dose-response curves in which the Prl concentration of the medium was plotted as a function of dopamine concentration. The slope of the dose-response curve for pituitaries from day 12 or pregnancy was significantly steeper than the slopes of the curves for pituitaries from days 5 and 18, which did not differ from each other. These data suggest that the Prl secretion mechanism is more sensitive to inhibition by dopamine on day 12 of pregnancy, when serum Prl concentration is very low, than on days 5 or 18, when serum Prl concentrations are higher. One of the mechanisms by which circulating Prl concentrations are reduced to very low levels during midpregnancy in the mouse may be increased pituitary sensitivity to dopamine.

Hypophysectomy of pregnant mice on days 10, 12, and 16 of gestation resulted in a large and rapid (12-24 h postoperatively) increase in serum placental lactogen (mPL) concentrations, as measured by homologous RIA. Serum progesterone (P4) concentrations decreased compared to those in intact and sham-operated controls. Significantly, the increase in serum mPL in mice hypophysectomized on days 10 and 12 preceded any decrease in serum P4 concentrations by at least 24 h, suggesting that two indirectly related mechanisms may be involved in the control of the serum concentrations of these hormones. The weight of the uterus, including the feto-placental units, placental weight, and placental content of mPL did not differ between intact, sham-operated, and hypophysectomized groups. The gestational profiles of serum GH and PRL concentrations were similar in the sham-operated and intact controls. Although serum P4 concentrations decreased after hypophysectomy on days 10 and 12 of gestation, the maintenance of the pregnancy was not affected; thus, the reduced concentrations of serum P4 (approximately 50% of control by day 16) were adequate to allow the pregnancy to continue. Hypophysectomy on day 16 of gestation resulted in spontaneous abortion 48 h postoperatively in all but one animal. We conclude that the absence of some pituitary factor(s) results in a rapid increase in serum mPL, while a concomitant, albeit gradual, decrease in serum P4 begins 48 h postoperatively. Therefore, when the pituitary no longer contributes to the luteotropic complex of the midpregnant mouse, an increase in serum mPL alone cannot maintain serum P4 concentrations at control values.

The lactogenic response of mouse mammary gland explants to human placental lactogen (hPL) and ovine pituitary prolactin (oPRL) was examined on days 10 to 18 of pregnancy by measuring 3H-amino acid incorporation into calcium-rennin precipitable casein. To determine the lactogenic response of the explants, the mean slopes of dose-response curves were calculated for each hormone treatment. Slope means of dose-response curves for oPRL and hPL did not differ from each other on any day of pregnancy examined. A triphasic pattern of response was suggested when slope means of dose-response curves for both hormones were plotted as a function of day of gestation. Peak responses were observed on days 10, 13 and 17-18. Combinations of oPRL and hPL, in ratios of oPRL:hPL = 2:1 and oPRL:hPL = 1:2, also produced a triphasic pattern of sensitivity very similar to that produced by either hormone alone. These results suggest that mouse mammary explants may be more sensitive to oPRL and hPL on days 10, 13 and 17-18 of pregnancy.

The binding of recombinant mouse placental lactogen-I (mPL-Ir) to liver and ovarian membranes was investigated in virgin and pregnant mice. Competitive binding assays demonstrated that mPL-Ir, mouse placental lactogen-II (mPL-II), and mouse PRL (mPRL) bind to the same receptors in ovarian membranes. The relative abilities of the three hormones to displace [125I]iodo-mPL-Ir from the ovarian lactogen receptors was mPL-II greater than mPL-Ir much greater than mPRL. Scatchard analysis of mPL-Ir binding to ovarian membranes from day 10 pregnant mice showed a Ka of 2.0 x 10(9) M-1 and a binding capacity of 3.2 x 10(-14) mol/mg membrane protein. The specific binding of [125I]iodo-mPL-Ir to ovarian membrane preparations was significantly higher on day 17 than on day 10 of gestation. Dissociation of endogenous hormones with 4 M MgCl2 increased the binding of [125I]iodo-mPL-Ir to ovarian membranes but not to liver membranes. Affinity cross-linking of [125I]iodo-mPL-Ir to liver and ovarian membranes resulted in the specific labeling of proteins with receptor mol wt (Mr) of 44K and 40K (nonreduced) and 50K and 42K (reduced), as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The lactogen receptors from liver and ovary appeared structurally homologous, producing fragments with similar Mr when treated with proteolytic enzymes and undergoing similar reductions in Mr when treated with glycolytic enzymes. The ability of mPLs to bind specifically and with high affinity to receptors in mouse ovarian membranes suggests that these hormones may regulate ovarian function during gestation.