Progesterone, but not progesterone-independent activation of progestin receptors by a mating stimulus, rapidly decreases progestin receptor immunoreactivity in female rat brain.

Recent studies suggest that progestin receptors may be activated in vivo by neurotransmitters in the absence of ligand. More specifically, vaginal-cervical stimulation (VCS) can influence sexual behavior by activating progestin receptors in the absence of progesterone. Another way to test if progestin receptors are influenced by particular stimuli is to examine progestin receptor immunostaining. We report that progestin receptor immunoreactivity is decreased in the forebrain of estradiol-primed ovariectomized (OVX) rats within 1 h after a subcutaneous injection of progesterone, a time by which rapid down-regulation of progestin receptors does not seem to have occurred. In estradiol-primed OVX rats, VCS also decreased progestin receptor immunoreactivity within 1 h in the medial preoptic area, but not in any other area examined. To determine if the decrease in immunoreactivity by VCS was due to adrenal secretions or by ligand-independent activation of progestin receptors, we repeated the experiment in estradiol-primed OVX/adrenalectomized rats. Prior removal of the adrenal glands blocked the rapid decrease in progestin receptor immunoreactivity, even though data from other experiments suggest that progestin receptors are activated by VCS at this time. These studies suggest the possibility that progestin receptors may be affected differentially by progesterone-dependent or by progesterone-independent pathways. This raises the possibility that activation of progestin receptors by these two distinct pathways may lead to different neuronal consequences.

Castration does not inhibit aggressive behavior in adult male prairie voles (Microtus ochrogaster).

The relationship between castration and reduced male aggression is well established. However, anecdotal observations of male prairie voles (Microtus ochrogaster) suggest that castration does not reduce aggressive behavior. To investigate the role of testicular androgens on aggressive behavior, castrated or gonadally intact male prairie voles were paired in a neutral arena with a gonadally intact vole. Castration did not reduce the frequency of intermale aggression. In Experiment 2, aggressive behavior was examined further using resident-intruder, grouped aggression, and aggression against a lactating female models. Again, castration did not affect the frequency of aggression in male prairie voles. Taken together, the results of this study suggest that aggressive behavior may be independent of gonadal steroid hormones in adult male prairie voles.

Induction of progestin receptors by estradiol in the forebrain of estrogen receptor-alpha gene-disrupted mice.

Mice, rats, and humans have two types of estrogen receptors, estrogen receptor-alpha (ERalpha) and estrogen receptor-beta (ERbeta). Estrogen receptor-alpha gene-disrupted (ERalpha-disrupted) mice bear two nonfunctional copies of the ERalpha gene. This mutation blocks the synthesis of full-length ERalpha, renders the animals infertile, and inhibits the induction of female sexual behaviors by estradiol and progesterone. It is likely that many of the processes contributing to the regulation of sexual receptivity by estradiol and progesterone are compromised in ERalpha-disrupted mice. However, given the importance of progesterone in the regulation of sexual receptivity and given the importance of progestin receptors (PRs) in mediating the responses of females to progesterone, we investigated the effects of ERalpha disruption on the induction of PRs by estradiol in the forebrain. We hypothesized that estradiol would induce PRs in wild-type mice but not in ERalpha-disrupted mice. Ovariectomized wild-type and ERalpha-disrupted mice were implanted with either estradiol-filled capsules or empty capsules for 5 d, after which their brains were processed for the immunocytochemical detection of PR. Estradiol increased the number of PR-immunoreactive cells in both wild-type and ERalpha-disrupted mice. The residual responsiveness of ERalpha-disrupted mice to estradiol could be accounted for by an ERbeta-dependent mechanism or another as yet unidentified estrogen receptor; however, because ERalpha-immunoreactivity and PCR product representing the 3' end of ERalpha mRNA were found in at least one PR-containing region of the ERalpha-disrupted mice, an ERalpha splice variant may also mediate the induction of PR-immunoreactivity in ERalpha-disrupted mice.

Mating-related stimulation induces phosphorylation of dopamine- and cyclic AMP-regulated phosphoprotein-32 in progestin receptor-containing areas in the female rat brain.

Vaginal-cervical stimulation induces a number of physiological and behavioral events, including the facilitation of mating behavior. Although the facilitation of one component of mating behavior, lordosis, by vaginal-cervical stimulation does not require the presence of progesterone, it appears to be mediated by neural progestin receptors. Abundant evidence suggests that dopamine may play a role in the neural circuitry activated by vaginal-cervical stimulation, including the mating-induced release of dopamine in progestin receptor-containing areas of the brain, changes in the activational state of progestin receptors because of dopamine D1 receptor stimulation, facilitation of lordosis by D1 receptor stimulation in estradiol-primed rats via progesterone-independent events, and D1 agonist-induced neuronal responses in progestin receptor-containing areas and cells. We tested the hypothesis that vaginal-cervical stimulation induces phosphorylation of dopamine- and cyclic AMP-regulated phosphoprotein (DARPP-32; Mr = 32,000), a protein phosphorylated predominantly in response to the stimulation of D1 receptors. At 9 d after ovariectomy, female rats were injected subcutaneously with a behaviorally effective dose of estradiol benzoate. At 48 hr later they received vaginal-cervical or control (perineal) stimulation, and they were perfused 1 hr later. Vaginal-cervical stimulation increased the number of cells expressing pDARPP-32 immunoreactivity by 92% in the medial preoptic nucleus, 134% in the caudal ventromedial hypothalamic nucleus, 123% in the posterodorsal medial amygdala, and 103% in the bed nucleus of the stria terminalis. These results suggest that some of the neuronal effects of vaginal-cervical stimulation, and perhaps other social or environmental stimuli, are mediated by phosphorylation of DARPP-32, perhaps via stimulation of D1 receptors, within progestin receptor-containing areas.

Stress affects corticosteroid and immunoglobulin concentrations in male house mice (Mus musculus) and prairie voles (Microtus ochrogaster).

Glucocorticoids, secreted in response to perceived stress, can suppress immunoglobulin (Ig) levels and compromise immune function in mice and rats. Prairie voles (Microtus ochrogaster) have been reported to exhibit basal corticosterone concentrations that would cause pathological changes in the immune function of most other rodents. The goals of the present study were to verify that serum corticosterone concentrations are high in prairie voles, as compared with house mice (Mus musculus), by measuring serum corticosterone with the same RIA; to examine the effects of mild stressors on corticosterone response in both species and to examine the effects of elevated corticosterone levels on IgM and IgG levels in prairie voles and house mice. After 2 weeks of randomly timed 15-min daily restraint or cold-water swim sessions, animals were injected with sheep red blood cells. The data confirmed that basal blood concentrations of corticosterone were higher in prairie voles than house mice, but these high levels doubled after the first swim session in prairie voles, indicating that the adrenals can respond to stressors by producing increased corticosterone. After stress, antibody production (both IgM and IgG) was reduced in house mice but not in prairie voles, despite higher blood concentrations of glucocorticoids in prairie voles. Although body mass was statistically equivalent between species, prairie voles and mice differed dramatically in adrenal and splenic masses. Average adrenal mass of prairie voles was approximately three times the average mass of these organs in house mice; in contrast, the average splenic mass of house mice was approximately three times that of prairie voles. These data may be relevant to seasonal changes in immune function and survival.

The effects of photoperiod and food intake on reproductive development in male deer mice (Peromyscus maniculatus).

Seasonal breeding is a tactic that has evolved in rodents that limits reproduction to specific times of the year to increase reproductive success. In order to time breeding accurately, many animals respond to changes in daily photoperiod. Short day lengths inhibit breeding in many nontropical rodent species. Restricted food availability can also inhibit reproductive function among some individuals in these so-called "photoperiodic" populations. Rodents born at the end of the breeding season typically delay sexual maturation until the following spring. Prepubertal rodents exposed to day lengths that are < 12 h light/day will not undergo puberty for 4-7 months in the laboratory. Food restriction can also affect the timing of puberty onset. Reproductive function of food-restricted juvenile mice may remain inhibited until food availability improves. Alternatively, reproductive function of food-restricted juvenile mice might eventually develop despite restricted food intake. This study examined the effects of chronic food restriction and photoperiod on reproductive development in male deer mice (Peromyscus maniculatus bairdii). Short-day mice fed ad lib delayed gonadal development for 5-7 months, but eventually achieved reproductive maturity. The reproductive function of short-day mice fed ad lib was indistinguishable from long-day control animals when assessed at week 32. Long-day food-restricted mice exhibited an intermediate level of gonadal development and function. Short-day food-restricted deer mice also inhibited reproductive growth, but failed to demonstrate reproductive maturity by week 32 of the study. Taken together, these results suggest that retardation of reproductive development by food restriction is only superficially similar to the delay in reproductive maturation imposed by short day exposure. It does not appear that male deer mice escape from the inhibitory effects of food restriction to attain sexual development.

Progesterone-independent activation of rat brain progestin receptors by reproductive stimuli.

Activation of steroid hormone receptors by steroid hormones alters both the physiology and behavior of animals. Steroid hormone receptors (e.g., progestin receptors) can also be activated in the absence of steroid hormones by pharmacological treatment with neurotransmitters or neuropeptides. However, it is not known if progesterone-independent activation of brain progestin receptors occurs under natural, physiological, conditions. We report that increases in reproductive behavior and brain immediate early gene expression in female rats induced by mating stimuli can be blocked by prior treatment with progesterone antagonists in the absence of circulating progesterone. This suggests that progestin receptors are activated in a progesterone-independent manner by a physiologically relevant stimulus in female rats, thus implicating a novel pathway by which mating stimuli and other environmental influences could activate steroid receptors to influence neuronal response and behavior.

Reproductively-relevant stimuli induce Fos-immunoreactivity within progestin receptor-containing neurons in localized regions of female rat forebrain.

An experiment was conducted to determine if neurons that respond to stimuli associated with mating in female brain also contain progestin receptors. We found that a portion of the neurons that respond to stimuli associated with mating also contains progestin receptors. While the appropriate hormonal conditions are important for sexual receptivity, somatosensory information provided by the male also influences sexual behavior. One important stimulus provided by the male during copulation is vaginal-cervical stimulation (VCS). VCS has been shown to elicit many different behavioral and endocrine changes in female rats, such as increases in lordosis, pseudopregnancy, and termination of sexual receptivity. VCS also increases the expression of the immediate early gene product, Fos, in areas associated with reproduction. A portion of the neurons responding to VCS with increased Fos-immunoreactivity (Fos-IR) in female rat forebrain also contains estrogen receptors, illustrating that hormonal and mating-stimuli converge in a population of cells. As progesterone also plays an important role in female sex behavior, it is important to determine if some of the neurons also integrate information concerning serum progesterone levels and social interactions. Thus, we used a dual immunofluorescent technique to label both Fos-IR and progestin receptor-immunoreactivity (PR-IR) in the brains of estradiol-primed, ovariectomized female rats following VCS manually applied by the experimenter. Many of the neurons that respond to VCS with increased Fos-IR within the medial preoptic area, the arcuate nucleus, and the progestin receptor-rich areas of the rostral and caudal ventromedial nucleus of the hypothalamus also contain PR-IR.

Photoperiod and population density interact to affect reproductive and immune function in male prairie voles.

Seasonal breeding of rodents is often associated with changes in adrenal function; altered adrenal function could account, in part, for seasonal changes in immune function and, ultimately, influence seasonal fluctuations in survival. Animals commonly monitor the annual change in photoperiod to ascertain the time of year and to make appropriate seasonal adjustments in physiology and behavior. Several extrinsic factors affect reproductive responsiveness to photoperiod. The interaction between population density and reproductive and adrenal responsiveness to photoperiod was assessed in the present experiment. Adult male prairie voles (Microtus ochrogaster) were maintained individually for 10 wk in long [light:dark (LD) 16:8] or short (LD 8:16) photoperiods in rooms with either high (10.96 animals/m3) or low (0.18 animals/m3) population densities. Regardless of population density, short-day voles regressed the size of their reproductive organs; reproductive organ masses were higher in long-day voles housed in high-density compared with low-density rooms. Paired adrenal masses were reduced in short-day voles, but were unaffected by population density; serum corticosterone concentrations were significantly elevated in short-day compared with long-day animals. In both photoperiods, basal blood corticosterone levels were higher in voles from low-density compared with high-density rooms. Splenic masses were unaffected by day length, but were elevated among high-density animals. Similarly, serum immunoglobulin (IgG) levels were elevated among high-density animals. These results suggest that population density per se, in the absence of behavioral interactions, can affect reproductive size, and possibly function, in long-day conditions, and that prairie voles, which are highly social, exhibit higher corticosterone and lower IgG levels in low compared with high densities. These results may be important in understanding arvicoline population fluctuations, as well as improving animal husbandry practices in the lab.

Photoperiodic effects on steroid negative feedback in female prairie voles (Microtus ochrogaster).

Breeding in prairie voles is mainly restricted to the autumn and winter of most years. The organization of estrus in female prairie voles is unusual because behavioral estrus is induced by chemosensory stimuli from the urine of adult conspecific males. Isolated females exhibit undetectable levels of estradiol and never display estrous behavior, yet exposure to male urine causes a cascade of endocrine changes that evoke estrogen secretion from the ovaries and estrous behavior within 24 hr. In the prairie vole, the extreme dependence of estrus on chemosensory stimuli raises the possibility that their ovaries may be less prominent in the regulation of gonadotropin secretion than in species with more endogenously organized estrous cycles. The present study examined the contribution of the ovaries in luteinizing hormone (LH) regulation in prairie voles. Females were maintained for 9 weeks in either long (LD 16:8) or short (LD 8:16) photoperiodic conditions, a blood sample was obtained, and then animals were either ovariectomized or received a sham procedure. Another blood sample was obtained a week later and assayed for serum LH. Blood serum LH levels were significantly reduced in short-day voles, compared to long-day animals. After ovariectomy both long-day and short-day voles exhibited equivalent elevations in LH levels. This study provides evidence that photoperiod is measured in female voles and the ovaries appear to produce sufficient steroids to suppress LH release.

The effects of photoperiod on olfactory c-fos expression in prairie voles, Microtus ochrogaster.

Reproduction stops among the majority of prairie voles (Microtus ochrogaster) during the winter. Short day lengths suppress male reproductive function dramatically in the laboratory, but photoperiod exerts only subtle effects on female reproductive function. Thus, the regulation of seasonal breeding in this species remains partially unspecified. In contrast to commonly studied rodents, female prairie voles do not undergo spontaneous estrous cycles; rather, they are induced into estrus by exposure to chemosignals expressed in conspecific male urine. In the present study, the hypothesis was tested that seasonal breeding among female prairie voles in the field reflects photoperiod-mediated changes in the responsiveness of the chemosensory system to male urine. Responsiveness was assessed by localizing the product of the c-fos immediate early gene with an immunocytochemical procedure. Female prairie voles were maintained in either long (LD 16:8) or short (LD 8:16) photoperiods from birth until adulthood, and exposed to either male urine or skim milk. Immunocytochemistry for fos protein revealed an increased number of immunoreactive cells within the accessory olfactory system of female prairie voles, including the accessory olfactory bulbs, granule cell layer, as well as the medial and cortical divisions of the amygdala 1 h after exposure to a single drop of urine as compared to individuals exposed to skim milk. The number of immunoreactive fos cells induced in females by conspecific male urine was also affected by photoperiod; short day females displayed fewer immunoreactive fos neurons in the accessory olfactory system as compared to long-day animals.(ABSTRACT TRUNCATED AT 250 WORDS)

Reproductive and nonreproductive responsiveness to photoperiod in laboratory rats.

Laboratory rats (Rattus norvegicus) have been traditionally considered nonphotoperiodic because reproductive function is unaffected by day length. However, at least three experimental manipulations of rats--perinatal androgen injection, peripubertal androgen implants, and peripubertal olfactory bulbectomy--have been reported to unmask reproductive responsiveness to photoperiod. The physiological means by which early testosterone treatment or olfactory bulbectomy affect the expression of photoperiodism were hypothesized to operate through similar underlying mechanism(s) that involved gonadotropin and prolactin blood levels. Short day lengths reduce blood levels of gonadotropins in so-called photoperiodic rodent species. Reduced prolactin levels result in virtually all reproductively photoperiodic species housed in short day lengths. In Experiment 1, male weanling rats either were olfactory-bulbectomized or received a sham-procedure and housed for 10 weeks in long (LD 16:8) or short (LD 8:16) days. Short-day rats reduced body mass, testicular sperm counts, and the size of their reproductive systems; olfactory bulbectomy amplified this inhibitory effect for some parameters including testicular and epididymal sperm counts. However, neither short days nor olfactory bulbectomy affected blood titers of follicle stimulating hormone (FSH) or prolactin. Pelage density was also unaffected by photoperiod, but rats retained their juvenile fur color; i.e., short-day rats remained white, but long-day rats became yellowish. In Experiment 2, male rats were injected with testosterone at 3 days of age, then housed in long or short days until 10 weeks of age. Day length alone did not affect any experimental parameter measured in Experiment 2 except fur color; again, short-day rats retained their juvenile fur color.(ABSTRACT TRUNCATED AT 250 WORDS)

Proenkephalin transgenic mice: a short promoter confers high testis expression and reduced fertility.

The regulation and possible function of the preproenkephalin gene in testis were studied in vivo in transgenic mice containing: (1) bases -193 to +210 of the human proenkephalin gene and an additional one kilobase of 3' proenkephalin flanking sequence driving expression of bacterial chloramphenicol acetyltransferase (CAT), and (2) the same promoter and flanking sequences driving expression of a rat proenkephalin cDNA. Five lines of mice, designated HEC1-5, expressed the first construct and 10, HER1-10, the second. Each HEC male and many HER males showed dramatic expression of the transgene in the testis, although much lower expression was observed in the brain and other enkephalin-producing tissues. High levels of expression in testis can thus be achieved with a very short promoter region and do not require intron A sequences previously considered necessary. Altered enkephalin expression may affect testicular function. One founder, HER8, displayed grossly abnormal testicular morphology and was completely infertile. A second founder, HER6, had low sperm motility. Two offspring from other lines also displayed subnormal fertility. These studies support a role for specific promoter sequences in testis expression and may further support a significant role for proenkephalin in testicular function.

Day length influences proceptive behavior of female prairie voles (Microtus ochrogaster).

Prairie voles typically do not breed during the winter; instead, they typically confine breeding to the spring and summer. One proximate cue contributing to this seasonal change in breeding among males is photoperiod. However, photoperiod does not appear to affect female fecundity. Female prairie voles are induced into estrus by chemosensory cues in the urine of male conspecifies. Females are exposed to these cues when they groom the ano-genital region of males. Females that do not groom the ano-genital region of males rarely become estrus. Ano-genital grooming can be considered a proceptive behavior because the onset of estrus is dependent upon females exhibiting this behavior. This study addressed whether or not photoperiod influenced the proceptive behavior of female prairie voles. The results indicated that females maintained in a long photoperiod groomed the ano-genital region of anesthetized stimulus males more frequently than females maintained in a short photoperiod. This change in proceptive behavior may contribute to the seasonal changes in breeding exhibited by prairie voles.

Winter adaptations of male deer mice (Peromyscus maniculatus) and prairie voles (Microtus ochrogaster) that vary in reproductive responsiveness to photoperiod.

Individuals of many nontropical rodent species restrict breeding to the spring and summer. Seasonal reproductive quiescence putatively reflects the energetic incompatibility of breeding and thermoregulatory activities. However, so-called "out-of-season" breeding occurs in virtually all rodent populations examined, suggesting that the incompatibility can be resolved. Both reproductive inhibition and development of energy-saving adaptations are mediated by environmental photoperiod, but some individuals do not inhibit reproduction in short days. In order to assess the costs and benefits of winter breeding, the present study examined the extent to which male prairie voles (Microtus ochrogaster) and deer mice (Peromyscus maniculatus) that maintained summer reproductive function in winter-simulated daylengths also maintained summer thermoregulatory adaptations. Circadian locomotor activity patterns, basal metabolic rate, capacity for nonshivering thermogenesis, nest building, body mass, and daily food consumption were compared among short-day (LD 8:16) regressed males, short-day (LD 8:16) nonregressed males, and long-day (LD 16:8) males. Short-day nonregressed deer mice resembled long-day conspecifics in terms of body mass and nest-building activities; however, the locomotor activity pattern of short-day nonregressed deer mice was similar to that of their short-day regressed conspecifics. Short-day nonregressed prairie voles had body masses similar to those of long-day conspecifics. Regardless of their reproductive response to photoperiod, short-day prairie voles reduced their daily food consumption and wheel-running activity, compared to long-day voles. These results suggest that winter breeding has energetic costs, most likely resulting from maintaining a "summer-like" body mass relative to that of reproductively regressed animals. These costs may be ameliorated to some extent by the reduction in locomotor activity and nest-building behavior emitted by short-day animals, regardless of reproductive response to short days. Thus, the occurrence of winter breeding may be the result of sufficient numbers of reproductively photoperiod-nonresponsive morphs in the population and sufficiently mild ambient conditions to permit survival of these larger animals.

Ontogeny of chick responses to maternal food calls in the Burmese red junglefowl (Gallus gallus spadiceus).

Burmese red junglefowl (Gallus gallus spadiceus) chicks were reinforced at 5 days old with a mealworm at the end of a runaway in the presence of a maternal food call. Two food calls were used, one made to a highly preferred food item (a mealworm) and one made to a less preferred food item (chick crumbs). Chicks initially approached the food call made to the mealworm faster regardless of reinforcement contingencies, but by the 3rd day of training chicks responded faster to the call that was reinforced. These results indicate that functional experience can modify preexisting response tendencies to food-calling stimuli. We discuss the implications of these results for the interpretation of published accounts that chickens use food calls deceptively.

Effects of photoperiod and 6-methoxy-2-benzoxazolinone on male-induced estrus in prairie voles.

Prairie voles (Microtus ochrogaster) are characterized as facultative breeders with higher rates of reproductive activities observed during spring and summer than autumn and winter. The environmental factors regulating seasonal breeding in this species remain unspecified. Short day lengths inhibit reproductive organ development in male prairie voles in the laboratory, but these males remain fertile and capable of siring offspring; female prairie voles have been reported to be reproductively unresponsive to day length in the laboratory. The organization of estrus in this species is unusual in that females never display the cyclic changes associated with estrus; rather, female prairie voles require chemosensory stimuli associated with fertile males in order to be induced into estrus. The plant compound, 6-methoxy-2-benzoxazolinone (6-MBOA), is present in vegetatively growing grasses and sedges and acts to trigger reproduction in other rodent species exposed to short days. It was suspected that 6-MBOA present in the laboratory diet may have overridden the effects of photoperiod on female prairie voles in previous laboratory studies. In the present study, the effects of 6-MBOA and photoperiod on estrus induction were examined. Beginning at Week 0, female prairie voles were housed in long (LD 16:8) or short (LD 8:16) photoperiods for 9 weeks, then implanted subcutaneously either with an empty Silastic capsule or one packed with 6-MBOA. A special diet, devoid of 6-MBOA, was available ad lib from Week 5 to the end of the study.(ABSTRACT TRUNCATED AT 250 WORDS)

Pelvic fractures crash vehicle indicators.

Fracture of the pelvis often results in major hemorrhage and even death in victims of motor vehicle trauma. In the hospital emergency department, diagnosing a fracture of the pelvis is difficult because the patient may be unconscious or disoriented, have multisystem injury, may not demonstrate bruising or other physical findings, and because the attendant bleeding is often retroperitoneal. This paper develops information from a diagnostic source that is currently almost unused: the vehicle. Accident statistics files from the National Accident Sampling System are used to relate pelvic fractures and dislocations with vehicle information. Collision type, occupant seating position, intrusion, entrapment, restraint use, age, and sex are analyzed as they bear upon the likelihood of pelvic fracture. Linear discriminant analysis is used to form simple equations that predict whether the patient will have a fracture or dislocation of the pelvis.