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Biomedical subjects

F Talamantes

Publications and source records attributed to F Talamantes.

At least 127 records · Page 7Linked to original sources

In vitro kinetics of synthesis and release of mouse placental lactogen.

The dynamics of the in vitro synthesis and release of mouse placental proteins and mouse placental lactogen (mPL) were examined by culturing explants from mouse placentae from days 11, 14, and 17 of gestation for up to 24 h. The amount of mPL in medium and tissue was determined by RIA, and new protein synthesis was measured by the incorporation of [3H]leucine into mPL and proteins. The rate of synthesis of 3H-labeled proteins and [3H]mPL was constant for 17 h of incubation, confirming the viability of the placental tissue. Rapid and preferential release of newly synthesized mPL was observed; 88% of the mPL synthesized in 24 h was released into the medium, and the ratio of [3H]mPL to 3H-labeled proteins in the medium (14-18%) was significantly higher than that in the tissue (1-4%). There is not a stable storage pool of mPL under these conditions, as demonstrated by the similarity of the ratios of counts per min of [3H]mPL to nanograms of mPL in the medium and tissue. This conclusion is supported by the observed parallelism of mPL synthesis and release and the low tissue content of mPL. Release of mPL per mg tissue was similar for day 11 and day 14 placentae. It appears that mPL release in the absence of in vivo regulatory factors is strictly a function of placental mass at this time in gestation. A lower release from cultured day 17 placentae probably represents a decrease in the level of metabolic activity of the placenta as parturition approaches.

Animals↗

The sensitivity of prolactin secretion to dopamine changes during pregnancy in mice.

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.

Animals↗

Lactogenic response of cultured mouse mammary epithelial cells to mouse placental lactogen.

The ability of mouse placental lactogen (mPL), mouse prolactin (mPRL), mouse GH (mGH) and ovine prolactin (oPRL) to stimulate synthesis of alpha-lactalbumin was tested in a primary culture of mouse mammary gland epithelial cells. Mammary tissue was obtained from 10-day pregnant Swiss Webster mice, enzymatically dissociated and the cells were cultured on floating collagen gels for 5 days. The basic culture medium consisted of Nutrient Mixture F12/Dulbecco's Modified Eagle's Medium (1:1, v/v), containing 10 mg insulin/1, 5 mg cortisol/l, 10 micrograms epidermal growth factor/l, 5 g bovine serum albumin/l and 50 mg gentamycin/l. Mouse PL, mPRL, mGH and oPRL were added to the basic medium in concentrations from 1 microgram/l to 1 mg/l. Accumulation of alpha-lactalbumin in the culture medium was measured. For that purpose, mouse alpha-lactalbumin was purified from mammary tissue obtained from lactating Swiss Webster mice and a radioimmunoassay was developed. Mouse PL, mPRL and oPRL stimulated a dose-dependent increase in alpha-lactalbumin secretion. Mouse GH also caused a significant, but dose-independent, increase in alpha-lactalbumin secretion. Mouse PL showed the greatest activity in stimulating alpha-lactalbumin secretion. It was concluded that mPL is an important lactogenic hormone in the latter half of pregnancy in the mouse, when circulating mPRL concentrations are low.

Animals↗

Nucleotide sequence of mouse prolactin and growth hormone mRNAs and expression of these mRNAs during pregnancy.

The mRNAs for mouse prolactin and growth hormone have been isolated from anterior pituitary glands and cloned as cDNAs. The nucleotide sequences of these mRNAs have been determined, and these sequences, along with the predicted amino acid sequences, are compared to those of other mammalian prolactin and growth hormone mRNAs. Levels of prolactin and growth hormone mRNAs during pregnancy have been monitored by hybridization to the cloned cDNA probes. We find the levels of these mRNAs to remain nearly constant during mid-to-late gestation.

Amino Acid Sequence↗

Placental lactogen secretion in the mouse: in vitro responses and ovarian and hormonal influences.

In vivo and in vitro methodologies were utilized to study the regulation of placental lactogen (PL) secretion in the mouse. Chorioallantoic placental explants obtained from midpregnant (day 10 of gestation) C3H/HeN or Swiss Webster mouse conceptuses behaved differently in culture than explants of chorioallantoic placentas obtained later in pregnancy. Midpregnant placental explants maintained or showed an increasing amount of PL output per day for 5-6 days of culture, while gestationally older placental explants demonstrated a progressive decline in PL output per day over the 5-6-day culture period. On selected days of gestation, bilaterally ovariectomized C3H/HeN or Swiss Webster mice were shown to have significantly higher serum PL concentrations than sham operated mice. Reduction of serum progesterone concentrations by unilateral ovariectomy and/or partial lutectomy had no significant effect on serum PL concentrations. Progesterone significantly reduced PL production by midpregnant placental explants from both C3H/HeN and Swiss Webster mice, whereas 17 beta-estradiol did not significantly affect PL release in vitro. PL production by mouse chorioallantoic placentas has gestational specific characteristics in vitro and is responsive to regulatory influences of the maternal ovaries and progesterone.

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Identification of proliferin mRNA and protein in mouse placenta.

Proliferin is a recently described, prolactin-related protein whose mRNA appears in several murine cell lines during active growth. We have surveyed a number of mouse organs or tissues for the presence of mRNAs that hybridize to cloned proliferin cDNA. Of the tissues tested, only the placenta yielded proliferin-related mRNA. This placental RNA is about 1 kilobase in length, increases sharply between days 8 and 10 of pregnancy, and then gradually declines through day 18. It is more abundant in RNA extracted from the fetal, compared to the maternal, part of the placenta. From a cDNA plasmid library prepared from poly(A)+ placental RNA, two types of proliferin-related clones were isolated, differing in intensity of hybridization to proliferin cDNA. By nucleotide sequence analysis, a strongly hybridizing clone was found to be nearly identical to the proliferin cDNA clone isolated from a library prepared from mRNA of a growing mouse fibroblastic cell line. Using an antiserum prepared against a synthetic proliferin fusion protein, we show that proliferin is secreted as a glycoprotein by minced placental tissue and that it differs from mouse placental lactogen. We conclude that proliferin is a placental hormone that is synthesized in certain mouse cell lines during active growth. Its function during pregnancy and during the growth of cultured cells is presently unknown.

Amino Acid Sequence↗

Ectopic pituitary grafts in mice: hormone levels, effects on fertility, and the development of adenomyosis uteri, prolactinomas, and mammary carcinomas.

Some endocrinological consequences of grafting hypophyses of mice to sites distant from the hypothalamic-pituitary portal vessels were investigated. Serum PRL levels in recipients rose within 3 weeks to levels seen during pregnancy, resulting in a premature increase in serum progesterone (P) levels. After 7 weeks, luteolytic effects were evident in BALB/c females, and P values had plateaued in the range of those seen in normal adult animals, while in BALB/c X C3H F1 hybrids, this effect was delayed, and P values rose, reaching, in some animals, levels reported during pregnancy. Despite continuing hyperprolactinemia, the fertility of graft-bearing females was reduced only slightly. By the 6th month, lesions of adenomyosis were frequent in uteri of C3H and F1 hybrids, but essentially absent from BALB/c animals. The number of corpora lutea was reduced in all experimental groups, yet serum P values were somewhat elevated in hybrid females and BALB/c animals bearing three grafts. Evidence is presented indicating that PRL enhances the action of P on both the estrogen-stimulating vagina and uterus. Small groups of grafted BALB/c and hybrid females not infected with exogenous mouse mammary tumor virus were allowed to survive until mammary cancers developed or until they reached 18 months of age. The grafts in the hybrids routinely became adenomatous, producing serm PRL levels of 1-21 X 10(3) ng/ml; only one secreted excessive GH. Adenomyosis wained in the aged hybrids, apparently due to declining ovarian function. The majority of the animals developed adenoid cystic mammary cancers that seemed to arise from areas of hyperplasia within dilated ducts rather than from alveolar hyperplasias, as is most frequently the case in mice carrying exogenous mouse mammary tumor virus.

Aging↗

Effects of different numbers of ectopic pituitary transplants on regulation of testicular LH/hCG and prolactin receptors in the hamster (Mesocricetus auratus).

Adult male hamsters were given transplants of 1/2, 1, 2, 3 or 4 pituitaries under the kidney capsule and were killed 4 weeks later. Pituitary transplants produced a significant, dose-related increase in plasma prolactin levels, no changes in plasma LH and an increase in plasma FSH. Concentration of LH/hCG receptors in the testes was significantly increased in animals with 2 or 3 transplants and concentration of testicular prolactin receptors was significantly increased in those given 2 transplants. The apparent stimulatory effects of 1/2, 1 or 4 transplants on testicular LH/hCG and prolactin binding were not statistically significant. Some of the animals were injected with 0.3 i.u. hCG/g body weight 24 h before being killed. This produced a significant reduction in the levels of prolactin receptors and an apparent reduction in the levels of LH/hCG receptors in the testes. Elevation of plasma testosterone concentrations in response to hCG was significantly greater in animals given 3 or 4 pituitary transplants than in the remaining groups. These results provide further evidence that prolactin increases the number of LH/hCG and prolactin receptors in the hamster testis and suggest that changing the number of ectopic pituitary transplants may result in biphasic effects on the testis, with 2 or 3 transplants being maximally stimulatory.

Animals↗

Regulation of testicular LH/hCG receptors in golden hamsters (Mesocricetus auratus) during development.

During prepubertal development in the golden hamster, there are major age-related changes in the number of testicular LH/hCG receptors. Between 22 and 35 days of age, there was greater than 10-fold increase in testicular LH/hCG receptors, followed by a decrease at Day 37. Concomitant with, but preceding slightly, the changes in receptors, were increases in plasma LH and FSH and most noticeably prolactin concentrations, between Days 10 and 20 of age. Inhibition of the increases in plasma levels of prolactin by daily injections of bromocriptine, between 14 and 31 days of age, resulted in suppressed testicular and seminal vesicle weights, and decreased content and concentration of testicular LH/hCG receptors. Similarly, the premature increase in plasma prolactin concentrations in prepubertal hamsters between 6 and 20 days of age, by means of ectopic pituitary transplants, resulted in increased testicular and seminal vesicle weights, as well as an increase in the concentration of testicular LH/hCG receptors. These results strongly suggest that increases in plasma prolactin values during development are important in enhancement of the development of testicular LH/hCG receptors.

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Effects of neonatal treatment with diethylstilbestrol and 17 alpha-hydroxyprogesterone caproate on in vitro pituitary prolactin secretion.

The effects of neonatal hormone treatment with diethylstilbestrol (DES) and 17 alpha-hydroxyprogesterone caproate (HPC) on days 1-5 of life on serum prolactin (PRL) levels and 3H-PRL synthesis and release were studied in C3H/MTV+ mice at 2, 4, 6, 8 and 10 weeks of age. Neonatal treatment of mice with 2.5 micrograms/day DES was the only treatment that affected the developmental pattern of serum PRL levels. Serum PRL levels were significantly decreased at 6 wks of age with this dose of DES. Neonatal treatment with 2.5 micrograms/day DES and 150 micrograms/day HPC affected the developmental pattern of H-PRL synthesis by the pituitary. At 10 wks of age 3H-PRL synthesis was significantly decreased by these doses of DES and HPC. The percent of 3H-PRL released did not differ between neonatally hormone treated and control animals, suggesting that neonatal treatment affected mechanisms that regulate PRL synthesis but not those that regulate release.

17-alpha-Hydroxyprogesterone↗

Neuroendocrine changes in male hamsters following photostimulation.

Transfer of gonadally regressed male golden hamsters from a short (5 L:19 D) to a stimulatory (14 L:10 D) photoperiod elicits, within 24 hr, significant changes in hypothalamic dopamine, serotonin, and possibly norepinephrine metabolism. Hypothalamic LHRH content was significantly elevated in short-photoperiod animals, but within 24 hr of transfer to a 14:10 photoperiod, LHRH declined to levels not different from those in hamsters maintained continuously in a long photoperiod. Plasma FSH levels were also significantly elevated within 24 hr of transfer, but increases in plasma LH were somewhat slower. Chronic treatment with the tyrosine hydroxylase inhibitor, alpha-methyl tyrosine (alpha MPT), which inhibits catecholamine synthesis, blocked the effect of a stimulatory photoperiod on plasma FSH levels, while treatment of 5:19 hamsters with the catecholamine precursor, L-dopa, mimicked the effects of photostimulation on plasma FSH levels. Testicular weights were not affected by alpha MPT or L-dopa treatment for 1 week. From these data, it appears that endocrine events associated with photoperiod-induced testicular recrudescence are under the control of hypothalamic neurotransmitters.

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Prolactin profiles during estrous cycle and pregnancy in hamster as measured by homologous RIA.

The development of a homologous radioimmunoassay (RIA) for the measurement of serum hamster prolactin (PRL) has facilitated our studies on monitoring the secretion patterns of this hormone during different states of reproduction. Four salient findings emerge from the present studies. First, rapid cardiac puncture without anesthesia does not affect serum PRL levels during selected times in the estrous cycle or during pregnancy. Second, serum PRL levels during the estrous cycle describe a daily rhythm of PRL release with maximum serum concentrations each day occurring in the afternoon and the highest of these occurring on proestrus (day 4 of cycle). Third, PRL release during pregnancy is characterized by a single surge each day; although the time of maximum PRL release varies, levels increase during the hours of darkness and decline to basal during the light. Fourth, the amount of PRL released during pregnancy diminishes with time; on day 15 the total amount of PRL in the serum is less than 10% of that measured on day 5.

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Immunocytochemical localization of mouse placental lactogen in the mouse placenta.

The localization of mouse placental lactogen (mPL) in the mouse placenta has been examined with the avidin-biotin-peroxidase complex (ABC) technique. Staining was observed to occur in the basal zone of placentae from days 10, 12, 15, and 18 of gestation. Within the basal zone, the most prominent staining was localized to the cytoplasm of the giant cells. Within the labyrinth, staining was observed in the giant cells in day-15 and day-18 placentae, with a greater number of stained cells being observed in the latter.

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Does prolactin modify testosterone feedback in the hamster? Pituitary grafts alter the ability of testosterone to suppress luteinizing hormone and follicle-stimulating hormone release in castrated male hamsters.

Adult male golden hamsters maintained in a long photoperiod (14 h of light and 10 h of darkness) or in a short photoperiod (5 h of light and 19 h of darkness for 7 weeks) were castrated and either given one anterior pituitary transplant under the kidney capsule or sham-operated. Additional animals were castrated and grafted or sham-grafted at the time of transfer to the short photoperiod. Starting 2 weeks after castration, all animals were injected three times a week with 20 micrograms testosterone propionate (TP). After 3 weeks, the dose of TP was increased to 80 micrograms and, after an additional 2 weeks, to 320 micrograms per injection. Blood samples were collected 2 weeks after castration and 1 day after the last injection of 20, 80, and 320 micrograms TP. Short photoperiod reduced and pituitary grafts increased plasma PRL levels. Plasma testosterone levels were related to the dose of injected TP, but were not influenced by photoperiod or pituitary transplants. Before the onset of TP injections, plasma LH and FSH levels in grafted and sham-grafted hamsters did not differ. In each of the three photoperiod conditions, injections of TP were consistently less effective in suppressing plasma gonadotropin levels in pituitary-grafted animals than in sham-grafted controls. These results indicate that PRL modulates the effects of exogenous testosterone on LH and FSH release in adult castrated male golden hamsters, this effect of PRL is due to reducing the sensitivity of the hypothalamic-pituitary system to feedback inhibition by testosterone, and suppression of pituitary PRL release in short photoperiod may be partially responsible for the concomitant increase in the sensitivity of LH and FSH release to inhibition by testosterone.

Animals↗

Does prolactin modify testosterone feedback in the hamster? Suppression of plasma prolactin inhibits photoperiod-induced decreases in testosterone feedback sensitivity.

The hormonal changes during the photoperiodically driven annual reproductive cycle of the male golden hamster can be explained partially by a change in the sensitivity of the hypothalamo-pituitary axis to negative feedback by testosterone (T). The present studies test the hypothesis that the increases in plasma levels of LH and FSH that follow photo-stimulation are due to decreasing feedback sensitivity and examine if this change in sensitivity is dependent upon increasing PRL levels. Adult males were exposed to a lighting schedule of 5 h of light, 19 h of darkness (5:19) for 12 weeks to induce gonadal regression. The animals were castrated; treated with an inhibitor of PRL release, bromocriptine (CB-154), or oil; and received a Silastic capsule that was empty or filled with T. Subsequently, the animals were transferred to 14:10 and killed 9 or 31 days later. There were no significant changes in FSH and LH in animals receiving oil injections and empty implants between days 9 and 31, suggesting no steroid-independent changes in gonadotropin secretion during this time period. However, 4-mm T implants were more effective in suppressing LH and FSH levels on day 9 than on day 31. This suggests that there is a gradual decrease in feedback sensitivity to T following photostimulation. T was more effective in inhibiting LH and FSH levels in CB-154-treated than in oil-treated animals on both day 9 and day 31. Thus, increases in PRL release are instrumental in causing decreases in feedback sensitivity following photostimulation.

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Differential effects of cryptorchidism and short photoperiod on testicular prolactin and LH/human chorionic gonadotrophin receptors in the golden hamster.

Gonadal atrophy induced by short photoperiods in male golden hamsters is accompanied by a decrease in the total content of testicular prolactin and LH/human chorionic gonadotrophin (LH/hCG) receptors and a concomitant rise in the concentrations of LH receptors. Atrophic testes are typically abdominal. In this study, we have compared the effects of experimentally induced cryptorchidism and the effects of a short photoperiod on testicular prolactin and LH/hCG receptors. Cryptorchidism produced the expected decrease in testicular weight and an increase in plasma FSH concentrations, without altering plasma concentrations of LH or prolactin and with a relatively small decrease in plasma testosterone. It also produced an increase in the concentration of prolactin and LH/hCG receptors without significant changes in their total content. Exposure to a short photoperiod decreased weights of testes and seminal vesicles and plasma levels of prolactin and testosterone, with no significant effects on plasma gonadotrophin levels. Short photoperiod also increased the concentration of LH/hCG receptors but, in contrast to the effects of cryptorchidism, it produced a precipitous decline in the total content of testicular receptors for both prolactin and LH/hCG. We conclude that changes in prolactin and hCG binding measured in membrane preparations from decapsulated testes of hamsters exposed to a short photoperiod cannot be explained by the abdominal position of the gonads, abnormalities of Sertoli cell function or changes in the relative proportion of Leydig cells in the atrophic testes.

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Pre-parturitional changes in serum prolactin, placental lactogen, growth hormone, progesterone, and corticosterone in the C3H/HeN mouse.

Pre-parturitional changes in serum prolactin, placental lactogen, growth hormone, progesterone, and corticosterone in the C3H/HeN mouse are described. Serum prolactin concentrations display an apparent biphasic pre-parturitional increase. Both serum placental lactogen and growth hormone concentrations are elevated during the second half of pregnancy. Serum placental lactogen concentrations remain elevated until parturition, whereas serum growth hormone concentrations decline on the last two days of pregnancy. Serum progesterone and corticosterone concentrations are elevated during the latter half of pregnancy and decline on the day preceding parturition.

Animals↗