Effect of hormones on primary cell cultures from pregnancy-dependent mouse mammary tumors.

No hormone combination was successful in maintaining long term primary cultures of pregnancy-dependent mammary tumors. Insulin provided the most consistent dose dependent stimulatory effect on short term proliferation as measured by 3H-TdR incorporation into DNA. Insulin in combination with corticosterone and prolactin produced the greatest stimulatory effect in most tumors. Both insulin at 5 microgram/ml and insulin, prolactin and hydrocortisone induced a partially synchronous wave of DNA synthesis in restimulated cultures. The time course of the wave of DNA synthesis was different for different hormone treatments. Insulin as 5 microgram/ml caused an earlier wave of DNA synthesis than insulin, prolactin plus corticosterone.     

Some endocrine traits of transgenic rabbits. I. Changes in plasma and milk hormones

The aim of these studies was to compare some endocrine and non-endocrine characteristics of transgenic (carrying mammary gland-specific mWAP-hFVIII gene construct) and non-transgenic rabbits. The concentrations of corticosterone, progesterone, testosterone, estradiol, insulin-like growth factor I (IGF-I) and human factor VIII (hFVIII) in the blood plasma of adult females (9 months of age, third generation transgenic animals), adult males, and young females (1-2 months of age, fourth generation of transgenic animals), as well as in the milk of lactating adult females, were analyzed by using RIA. In addition, litter size and body mass of pups born by transgenic and non-transgenic females from the third generation were compared. Transgenic animals were compared with their non-transgenic siblings (the same genetic and epigenetic background). Transgenesis did not influence plasma hFVIII, but significantly increased corticosterone (in all animals), reduced IGF-I (in adult males and females), testosterone and estradiol, (in young females) and altered progesterone (increase in adult males and decrease in adult females) concentrations in blood plasma. In addition, transgenic females had higher milk concentrations of testosterone, but not progesterone or IGF-I than their non-transgenic sisters. These endocrine changes were not associated with changes in litter size. Transgenic male (but not female) pups have smaller body mass than control animals. These observations demonstrate the influence of transgenesis per se on the animal growth and endocrine system (secretion of reproductive and stress steroid hormones as well as growth factors) over four generations.     

Reproduction in wild female olive baboons

The purpose of this paper is to evaluate several factors that influence female reproduction in a large troop of wild olive baboons (Papio cynocephalus anubis) based on 4 consecutive years of demographic data. Interbirth intervals were significantly shorter for females whose infants died before their next conception than for females whose infants survived. High-ranking mothers of surviving infants had significantly shorter birth intervals than comparable low-ranking mothers, independent of maternal age. This occurred mainly because the interval from resumption of cycling to conception was significantly shorter for high-vs. low-ranking females. Dominance rank did not influence sex ratio at birth, infant survival in the first 2 years, or adult female mortality. Age was also significantly related to interbirth intervals, with older females having shorter intervals. Primiparous females had consistently longer reproductive intervals than did multiparous females, but this difference reached statistical significance only for females whose infants died before the next conception. Primiparous females also experienced significantly higher infant mortality. Data on body size and estrous cycle length indicated no differences between high- and low-ranking females. Nutritional and stress-related mechanisms that may underlie the reproductive advantages of high rank are discussed.     

Morbidity and survival in neonates ventilated for the respiratory distress syndrome.

In a retrospective analysis the records of all (210) infants ventilated to treat the respiratory distress syndrome over three years were reviewed. A mortality of 19% was found. Intraventricular haemorrhage was associated than a significant increase in mortality in infants of less with 30 weeks'' gestation (p less than 0.001) and was the commonest cause of death. Pneumothoraces developed in one third of babies regardless of gestational age but were significantly associated with an increase in mortality only in infants of 27-29 weeks'' gestation. Patent ductus arteriosus was present in 31 infants and was commoner in babies of very low birth weight. The presence of a patent ductus arteriosus was not associated with decreased survival but was significantly related to an increased need for prolonged respiratory support (p less than 0.001). Thirty six infants developed chronic lung disease, three of whom died. Comparison with data from earlier studies indicated a steady improvement over the past decade in outcome for infants ventilated for the respiratory distress syndrome.     

Deaths in a desert baboon troop

Over a 4-year period, three of 22 infants born in a desert-dwelling chacma baboon troop (Papio ursinus) survived to more than 6 months of age. Tick infestation and infant kidnapping by adult females were the main causes of infant death. The high neonatal mortality resulted in a rapid birth rate through a decrease in duration of interbirth intervals for mature females. There were more adult male deaths in the troop than adult female deaths. Males died from canine inflicted wounds sustained during fights.     

Estrogen sensitive cell lines: Establishment and characterization of new cell lines from estrogen-induced rat pituitary tumors

Rat pituitary tumors were induced by the monthly injection of high doses of estradiol valerate. Of 12 animals which received the estrogen, 10 developed tumors. From these 10 tumors, 4 cell lines were successfully established and they have been maintained in culture for over 18 months. Several clones have been isolated from these established cell lines. Three of the 4 cell lines produce tumors in females considerably faster than in males or castrated females. Tumor-bearing animals have significantly increased amounts of rat growth hormone and prolactin in their serum. The 4 established cell lines, as well as clones derived from them, and tumors obtained in situ by injection of these cells growing in culture or successive transplants have shown the presence of an estrogen-binding protein of high affinity and low capacity. This estrogen-binding protein is similar to that described in other target organs (uterus, mammary glands, etc). These cell lines will be used to study the mechanism of action of estrogen in target cells as well as the synthesis and secretion of pituitary hormones.     

Sex-biased nestling mortality in the Montagu's harrier Circus pygargus

I evaluate causes and patterns of nestling mortality in a sexually dimorphic species, the Montagu's harrier Circus pygargus , and their relationship with sex and condition. Starvation was apparently the main reason for nestling death. Condition of birds that died was lower than those that survived. Both probability of nestling death and the proportion of nestlings that died within a brood increased with the number of hatched nestlings in a brood, and with increasing hatching date. For the nestlings that died after being sexed, when controlling for brood effects, probability of death was significantly related to nestling sex, with smaller males having a higher probability of dying. The probability of nestling death if hatched late in the season was relatively greater for males than for females. There was also a significant interaction between sex and hatching date on nestling condition: the decline in condition if hatched late in the season was steeper for males than for females. Males did not have a higher probability of death when having more sisters: neither the probability of brood reduction nor the proportion of nestlings that died were significantly related to within-brood sex ratio. Results suggest that mortality may partly result from sibling competition: females, being the larger sex, might be better able to compete for food within a brood than their male siblings. Additionally, smaller males may be less able to recover from periods of declining body weight.     

Reproductive parameters of female Japanese macaques: Thirty years data from the arashiyama troops,Japan

Over a 30-year period from 1954 to 1983, 975 live births were recorded for Japanese macaque females at the Iwatayama Monkey Park, Arashiyama, Japan. Excluding unknown birth dates, primiparous mothers gave birth to 185 infants (182 cases with age of mother known) and multiparous mothers gave birth to 723 infants (603 cases with age of mother known). The peak month of birth was May with 52.3% of the total births occurring during the period. Multiparous females who had not given birth the previous year did so earlier than multiparous females who had given birth the previous year and also earlier than primiparous females. Among the females who had given birth the previous year, females whose infant had died gave birth earlier than females who had reared an infant the previous year. The offspring sex ratio (1:0.97) was not significantly different from 1:1, and revealed no consistent association with mother's age. Age-fecundity exhibited a humped curve. The annual birth rate was low at the age of 4 years but increased thereafter, ranging between 46.7% and 69.0%, at between 5 and 19 years of age, but again decreased for females between 20 and 25 years of age. Some old females displayed clear reproductive senescence. The infant mortality within the first year of age was quite low (10.3%) and the neonatal (less than 1 month old) mortality rate accounted for 49.0% of all infant deaths. There was no significant difference between the mortality rates of male and female infants. A female's rank-class had no apparent effect on the annual birth rate, infant mortality, and offspring sex ratio. These long-term data are compared with those from other primate populations.     

Characteristic features of the reproduction of Koshima monkeys,Macaca fuscata fuscata: A summary of thirty-four years of observation

The reproductive data for Japanese monkeys,Macaca fuscata fuscata, which had been recorded for the 34 years from 1952 to 1986 on Koshima, were analyzed in terms of the influence of changes in artificial food supplies, the differences in reproductive success between females, the timing of births, and the secondary sex ratio. Koshima monkeys increased in number until 1971 when the population density was still small and artificial provisioning was copious. As described byMori (1979b), the severe reduction in artificial food supplies, which began in 1972, had an enormous deleterious effect on reproduction: the birth ratio of adult females of 5 years of age or more fell from 57% to 25%; the rate of infant mortality within 1 year of birth rose from 19% to 45%; primiparous age rose from 6 to 9 years old on average; and there was an increased death rate among adult and juvenile females. The prolonged influence of “starvation” may be seen in the significantly delayed first births of those females that were born just before the change in food supplies. When reproductive parameters are compared between the females who belonged to six lineages in the group during these periods, they were found to be rather consistent, although some individual differences can be recognized among females and subgroups. The apparent trend was that some of the most dominant females retained superior reproductive success while that of the second-ranked females has tended to diminish over the years since 1972. Such opposing trends were seen only in the most dominant lineage group and such a difference was not recognized among the females of other lineages. The difference in reproductive success is discussed in relation to both the different situations that arise because of the artificial food supplies and differences in feeding strategies. Multiparous females, after a sterile year, gave birth somewhat earlier than those who reared infants in the preceding year and, when artificial provisioning was intense, they tended to give birth a little earlier than during other periods. There is some evidence that the mortality of later-born infants was higher than that of earlier-born infants after 1972. However, this difference may not be responsible for the differential reproductive success of females since the timing of births did not differ among lineages. Furthermore, during the time when many females gave birth continuously, prior to 1972, the infant mortality did not differ with respect to the timing of births. The differences in infant mortality were not correlated with the reproductive history, parity or age of the mother, or with the sex of the infant. The secondary sex ratio varied by only a small amount, from slightly male-biased ratio (114: 100) when correlated with reproductive history, parity, age of mother, sex and survival ratio for preceding infants, timing of birth, and lineage of the female. Furthermore, the change in artificial food supplies did not cause any modifications of the secondary sex ratios, despite its enormous deleterious effect on reproduction. The secondary sex ratio of Japanese monkeys may not be influenced by the social factors mentioned.     

In vitro interaction of estrogen and prolactin on hormone-dependent rat mammory tumors (38511).

Hormone-dependent, regressing mammary tumors in rats were incubated in a chemically defined culture medium containing tritium labeled leucine at 37 degrees for 2 hr. The rate of tritium incorporation into the protein fraction by these tissues was measured with and without the addition of 17beta-estradiol and/or ovine prolactin into the medium.Tumor tissue responded to either estrogen or porlactin by a 6-7% increase in the rate of 3-H-leucine incorporation. When the incubation medium contained both the hormones, the increase in the rate of 3-H leucine incorporation was 19% which was significantly greater than the sum of the effect of each single hormone measured independelntly. These results demonstrate a synergistic effect between estrogen and prolactin on rat mammary tumors.     

Grooming relationships of adolescent orphans in a free-ranging group of Japanese macaques (Macaca fuscata) at Katsuyama: a comparison among orphans with sisters, orphans without sisters, and females with a surviving mother

The present study examined grooming relationships of adolescent females in a free-ranging group of Japanese macaques (Macaca fuscata) at Katsuyama. To assess whether the loss of the mother influenced the grooming relationships of adolescent females (5–7 years old), we compared the time spent in grooming interactions and the number of grooming partners among the following three groups: 6 adolescent orphans with sisters, 9 adolescent orphans without sisters, and 11 adolescent non-orphans with surviving mothers. In Japanese macaques, grooming most frequently occurs between mothers and their daughters. Therefore, it is expected that if the mother is lost, orphans will devote less time to grooming interactions than non-orphans. However, the time spent in overall grooming interactions did not differ among the three groups. While non-orphans maintained grooming relationships with their mothers, orphans acquired alternative grooming relationships with other group members. Orphans adopted two kinds of tactics to compensate for the loss of the mother. First, adolescent orphans with sisters developed more affiliative grooming relationships with their sisters than non-orphans with sisters. Secondly, adolescent orphans without sisters spent more time in grooming interactions with same-aged females and non-related adult females. Moreover, regarding grooming interactions with same-aged females and non-related adult females, orphans without sisters had a larger number of grooming partners than non-orphans. These results indicate that adolescent females have enough flexibility to develop their grooming network after the loss of their mothers, and that the lack of mother and sisters might accelerate socialization of adolescent females and enable them to be integrated in reciprocal adult grooming relationships.     

In vivo evidence for epidermal growth factor receptor (EGFR)-mediated release of prolactin from the pituitary gland

Members of the epidermal growth factor receptor (EGFR/ERBB) system are essential local regulators of mammary gland development and function. Emerging evidence suggests that EGFR signaling may also influence mammary gland activity indirectly by promoting the release of prolactin from the pituitary gland in a MAPK and estrogen receptor-α (ERα)-dependent manner. Here, we report that overexpression of the EGFR ligand betacellulin (BTC) causes a lactating-like phenotype in the mammary gland of virgin female mice including the major hallmarks of lactogenesis. BTC transgenic (BTC-tg) females showed reduced levels of prolactin in the pituitary gland and increased levels of the hormone in the circulation. Furthermore, treatment of BTC-tg females with bromocriptine, an inhibitor of prolactin secretion, blocked the development of the lactation-like phenotype, suggesting that it is caused by central release of prolactin rather than by local actions of BTC in the mammary gland. Introduction of the antimorphic Egfr allele Wa5 also blocked the appearance of the mammary gland alterations, revealing that the phenotype is EGFR-dependent. We detected an increase in MAPK activity, but unchanged phosphorylation of ERα in the pituitary gland of BTC-tg females as compared with control mice. These results provide the first functional evidence in vivo for a role of the EGFR system in regulating mammary gland activity by modulating prolactin release from the pituitary gland.     

Mother-infant relationships in bonnet macaques: Sources of variation in proximity

Family composition is an important predictor of variation in proximity among captive bonnet macaque (Macaca radiata) mothers and their infants. Infants that have several immature sisters in the group are initially more independent that infants with fewer sisters, but as they reach 6 months of age infants with several sisters become less independent than their peers. The effects of the presence of juvenile females on the relative independence of their infant siblings seem to be related to the fact that the members of families with several immature females are the targets of more aggression than are the members of other families.     

Effects of postnatal corticosterone treatment on reproductive development in the rat

Reproductive development was studied in rats in which corticosterone (25 mg/100 g body wt) mixed with an equal amount of cholesterol was implanted i.p. at Days 3, 6, 12 or 18 after birth. Vaginal perforation in rats treated on Days 12 or 18 was advanced by 10 days but reproductive function in the treated animals was generally disrupted. A variable incidence of persistent vaginal oestrus was observed in adult females given implants on Days 3 and 6, and of prolonged vaginal dioestrus in those treated on Days 12 or 18. All treated females had lordosis quotients lower than those of untreated controls, the effect being most pronounced at the earlier ages of treatment. The results are believed to indicate two periods in reproductive development which are differentially affected by corticosterone.     

Corticosterone alone does not trigger a short term behavioural shift in incubating female common eiders Somateria mollissima, but does modify long term reproductive success

The trade-off between reproductive effort and adult survival in birds is modulated by several factors. Corticosterone and prolactin have additive effects on reproductive behaviour by stimulating foraging and parental behaviours, respectively. When incubation is associated with fasting, nest desertion is supposed to be activated by an unknown refeeding signal when body condition becomes critically deteriorated. The concomitant rise in corticosterone levels has been suggested to be the triggering factor. We tested the role of corticosterone on reproductive success by observing the effect of corticosterone implants on reproductive success and on plasma prolactin concentration in female common eiders Somateria mollissima . Implanted females showed a significant increase in corticosterone and a decrease in prolactin levels. Despite their enhanced daily body mass loss, females did not abandon incubation nor did they start to refeed in the four days following implantation. These data show that the experimentally induced rise in plasma corticosterone concentration alone does not trigger nest desertion. However, after 25 days of incubation, implanted females displayed a higher rate of egg loss, suggesting lower nest attentiveness towards the end of incubation. We suggest that the short-term effects of corticosterone may be dependent on the energy state of the bird. However, the late-induced change in reproductive success is indirectly linked to corticosterone, and we suggest that either a prolactin decrease, or a depletion in protein body reserves, may participate in the long-term adjustment of incubation behaviour in female eiders.     

Effects of the MHC on hormonal induction of mammary tumors and function of hypophyseal isografts in the mouse

While the role of the H-2 complex in the resistance to virally induced tumors has been extensively studied, little is known about its influence on the development of epithelial tumors of non-viral etiology, although such tumors are most prevalent in humans. Therefore, we analyzed the role of the H-2 complex in susceptibility to mammary tumors induced by hormonal stimulation from heterotopic hypophyseal isografts in H-2 congenic strains from C57BL/10, BALB/c, and 020/A backgrounds. This method of induction allows an assessment of the effect ofH-2 genes on the function of various organs involved in this process. We found that the tumor susceptibility genes map to two segments: PE-S, and to the right of S. The mechanisms by which the H-2 complex affects the induction of mammary tumors in C57BL/10 congenic strains seem to include an influence on several factors involved in the hormonal stimulation, because the susceptible B10 congenic strains have higher plasma levels of prolactin and the H-2 complex also affects the growth of hypophyseal isografts. Their size correlates with tumor development in individual mice in the resistant C57BL/10 congenic strains. We reported previously H-2-dependent differences in levels of the estrogen receptor in hypophysis. For this study, we measured the levels of estrogen receptors in uteri to assess the tissue specificity of this effect of H-2. However, no influence of the H-2 complex on estrogen receptor levels was observed in uteri. Strains from BALB/c and 020 backgrounds developed mammary tumors much earlier than the B10 congenic strains, indicating a strong influence of non-H-2 genes.     

Lactation and care for unrelated infants in forest-living ringtailed Lemurs

An adult female ringtailed lemur (Lemur catta) known not to have been pregnant showed spontaneous lactation in response to twin infants born to an unrelated female. The females had met only 7 months earlier, when they and two other unrelated adult females were released from separate locations in a forest enclosure to form a new social group. Three months after release, an adult male from an adjacent enclosure gained access to the new group for 1 day, the day of one female's estrus. No males had access to the females throughout the remainder of the breeding season. Within 2 weeks of the birth of the twins, one of the other adult females began carrying the infants frequently, typically one at a time. All three females were checked for lactation when the infants were two months old. Both their mother and the unrelated adult who had been carrying the infants were producing milk. The third adult female, who never carried either infant, had no milk. The third adult female, who never carried either infant, had no milk. This female, however, like the two maternal females, frequently attacked unfamiliar immigrating adult males when the males approached the infants. Potential implications of these observations concerning the social organization of ringtailed lemurs are discussed.     

Evidence that corticosterone is not an obligatory intermediate in aldosterone biosynthesis in the rat adrenal

CGS 16949A is a potent inhibitor of aromatase in vitro with an IC50 of 0.03 microM for the inhibition of LH-stimulated estrogen biosynthesis in hamster ovaries. In vivo, CGS 16949A leads to sequelae of estrogen deprivation (e.g. regression of DMBA-induced mammary tumors) without causing adrenal hypertrophy in adult rats. To complement these in vitro and in vivo findings, the effect of CGS 16949A on adrenal steroid biosynthesis in rats was investigated in vitro and in vivo. The surprising finding in vitro was that CGS 16949A inhibited aldosterone biosynthesis (IC50 = 1 microM) at concentrations 100 times lower than those for inhibition of corticosterone biosynthesis (IC50 = 100 microM). Moreover, deoxycorticosterone (DOC) concentrations were elevated at all concentrations of CGS 16949A which inhibited aldosterone synthesis. The classical biosynthetic pathway for aldosterone is DOC----corticosterone----18-OH-corticosterone----aldosterone. Thus inhibition of aldosterone biosynthesis, reflected in DOC accumulation, without affecting corticosterone concentrations, indicates that corticosterone is not an obligatory intermediate in the conversion of DOC to aldosterone in the rat. In vivo, CGS 16949A showed a suppression of plasma aldosterone in ACTH-stimulated male rats at doses which did not significantly affect plasma corticosterone. In conclusion, aldosterone measured both in vitro and in vivo must be derived primarily from a biosynthetic pathway in which corticosterone is not obligatory intermediate.     

Interactions of insulin, corticosterone and prolactin in promoting milk-fat synthesis by mammary explants from pregnant rabbits

1. The rate of fatty acid synthesis by mammary explants from rabbits pregnant for 16 days or from rabbits pseudopregnant for 11 days was stimulated up to 15-fold by culturing for 2-4 days with prolactin. This treatment initiated the predominant synthesis of C(8:0) and C(10:0) fatty acids, which are characteristic of rabbit milk. 2. Inclusion of insulin in the culture medium increased the rate of synthesis of these medium-chain fatty acids. By contrast the inclusion of corticosterone led to the predominant synthesis of long-chain fatty acids. When explants were cultured for 2-4 days with insulin, corticosterone and prolactin, the rate of fatty acid synthesis increased up to 42-fold, but both medium- and long-chain fatty acids were synthesized. 3. These results show that the stimulus to mammary-gland lipogenesis and the initiation of synthesis of medium-chain fatty acids observed between days 16 and 23 of pregnancy in the rabbit can be simulated in vitro by prolactin alone. 4. When mammary explants from rabbits pregnant for 23 days were cultured for 2 days with insulin, corticosterone and prolactin, the rate of fatty acid synthesis increased fivefold, but there was a preferential synthesis of long-chain fatty acids. Culture with prolactin alone had little effect on the rate or pattern of fatty acids synthesized. 5. The results are compared with findings in vivo on the control of lipogenesis in the rabbit mammary gland, and are contrasted with the known effects of hormones in vitro on the mammary gland of the mid-pregnant mouse.