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Biomedical subjects

D Crews

Publications and source records attributed to D Crews.

At least 73 records · Page 4Linked to original sources
PubMed1996

Brain organization in a reptile lacking sex chromosomes: effects of gonadectomy and exogenous testosterone.

In mammals, males and females differ both genetically and hormonally, making it difficult to assess the relative contributions of genetic constitution and fetal environment in the process of sexual differentiation. Many reptiles lack sex chromosomes, relying instead on the temperature of incubation to determine sex. In the leopard gecko (Eublepharis macularius), an incubation temperature of 26 degrees C produces all females, whereas 32.5 degrees C results in mostly males. Incubation temperature is the primary determinant of differences both within and between the sexes in growth, physiology, and sociosexual behavior, as well as the volume and metabolic capacity of specific brain nuclei. To determine if incubation temperature organizes the brain directly rather than via gonadal sex hormones, the gonads of male and female leopard geckos from the two incubation temperatures were removed and, in some instances, animals were given exogenous testosterone. In vertebrates with sex chromosomes, the size of sexually dimorphic nuclei are sensitive to hormone levels in adulthood, but in all species studied to date, these changes are restricted to the male. Therefore, after behavior tests, morphometrics of certain limbic and nonlimbic brain areas were determined. Because nervous system tissue depends on oxidative metabolism for energy production and the level of cytochrome oxidase activity is coupled to the functional level of neuronal activity, cytochrome oxidase histochemistry also was performed on the same brains. Hormonal manipulation had little effect on the volume of the preoptic area or ventromedial hypothalamus in geckos from the all-female incubation temperature, but significantly influenced the volumes of these brain areas in males and females from the male-biased incubation temperature. A similar relationship was found for cytochrome oxidase activity of the anterior hypothalamus, amygdala, dorsal ventricular ridge, and septum. The only sex difference observed was found in the ventromedial hypothalamus; males showed no significant changes in cytochrome oxidase activity with hormonal manipulation, but females from both incubation temperatures were affected similarly. The results indicate that incubation temperature organizes the brain directly rather than via hormones arising from its sex-determining function. This is the first demonstration in a vertebrate that factors other than steroid hormones can modify the organization and functional activity of sexually differentiated brain areas.

Animals↗
PubMed1996

Effects of ovariectomy and estrogen replacement on attractivity and receptivity in the red-sided garter snake (Thamnophis sirtalis parietalis).

Activation of courtship behavior in male red-sided garter snakes, Thamnophis sirtalis parietalis, is independent of the presence of sex steroids. The only consistent treatment that stimulates courtship behavior in males is prolonged exposure to low temperature followed by subsequent warming, mimicking the emergence from hibernation. We investigated whether attractivity and receptivity in female red-sided garter snakes is similarly steriod independent. Female red-sided garter snakes are attractive when they emerge from hibernation and are courted by males; most mate within an hour of emergence. In a series of experiments, groups of females were either ovariectomized (OVEX) in the late spring, fall or while in hibernation. They were tested for attractivity and receptivity upon emergence from hibernation. Females OVEX in the spring were unattractive whereas those OVEX in fall or while in hibernation were attractive. Thus, attractivity appears determined the year before emergence and is dependent on the presence of the ovaries. All OVEX females were unreceptive upon emergence. OVEX females were also given replacement estradiol (E) treatment (either in Silastic capsules or single injections) at various points of their annual cycle. The only treatment that resulted in reinstating receptivity in OVEX females was the injection of E (20 micrograms) one hour prior to emergence. The effectiveness of E in reinstating receptivity was time dependent: the longer the period between emergence and injection, the less effective the same dosage was in stimulating receptive behavior. These experiments suggest that sexual behavior in female red-sided garter snakes is, unlike males, dependent on the presence sex steroid hormones. Although E is naturally at its lowest seasonal level upon emergence, the concentration is sufficient to stimulate receptivity. However, it appears that temperature regulates a time-limited window of sensitivity to E.

Animals↗
PubMed1996

Temperature and non-aromatizable androgens: a common pathway in male sex determination in a turtle with temperature-dependent sex determination?

This study addressed the hypothesis that, in the red-eared slider turtle, Trachemys scripta, non-aromatizable androgens are the physiological equivalent of temperature in determining male development. In the first experiment, eggs were treated in the middle of the temperature-sensitive period with 1.0 or 10.0 micrograms androsterone, 5 alpha-dihydrotestosterone, 3 alpha-androstanediol, or 3 beta-androstanediol, while at an all-male, male-biased, or one of two female-biased incubation temperatures. In the second experiment, eggs were treated with the same dosages of dihydrotestosterone at different stages of embryonic development while at a male-biased, threshold, or a female-biased incubation temperature. Results of experiment one indicated that hormone-induced masculinization is specific to non-aromatizable androgens. Results of experiment two indicated that the sensitivity to dihydrotestosterone corresponds to the temperature-sensitive window during development. Further, there is a dose-response relationship but no apparent synergism between exogenous dihydrotestosterone and incubation temperature. When considered with other research, it is suggested that non-aromatizable androgens and their products are involved in the initiation of male sex determination whereas oestrogens and their aromatizable androgen precursors are involved in the initiation of female sex determination.

Androstane-3,17-diol↗
PubMed1996

Temperature-dependent sex determination: the interplay of steroid hormones and temperature.

Sex determination is the product of coordinated gene expression. Mutational analyses have yielded great progress in our understanding of mammalian sex determination, and insight into the evolution of this sex chromosome system would be valuable. Mammals arose from turtle-like reptiles, and in many turtles the incubation temperature of the egg determines the sex of the offspring, a process known as temperature-dependent sex determination. There is mounting evidence that sex steroid hormones are the physiological equivalent of incubation temperature and serve as the proximate trigger for male and female sex determination. Temperature appears to accomplish this end by acting on genes coding for steroidogenic enzymes and sex steroid hormone receptors. The ability to manipulate sex determination in turtles both by temperature and by sex steroid hormones extends our understanding of the evolution as well as the physiology and molecular biology of sex determination.

Amino Acid Sequence↗
PubMed1995

Electrolytic lesions to the ventromedial hypothalamus abolish receptivity in female whiptail lizards, Cnemidophorus uniparens.

Lesions to the ventromedial hypothalamus (VMH) inhibit receptive sexual behavior in the whiptail lizard, Cnemidophorus uniparens, an all-female species. All lesions to the VMH that effectively abolished receptivity specifically damaged the dorsal lateral VMH, an area containing high concentrations of estrogen receptor in this species. These data further emphasize conservation of the VMH as a brain area critical to the expression of female sexual behavior in vertebrates.

Animals↗
PubMed1995

Incubation temperature and gonadal sex affect growth and physiology in the leopard gecko (Eublepharis macularius), a lizard with temperature-dependent sex determination.

Temperature-dependent sex determination (TSD), in which the temperature at which an egg incubates determines the sex of the individual, occurs in egg-laying reptiles of three separate orders. Previous studies have shown that the embryonic environment can have effects lasting beyond the period of sex determination. We investigated the relative roles of incubation temperature, exogenous estradiol, and gonadal sex (testis vs. ovary) in the differentiation of adult morphological and physiological traits of the leopard gecko, Eublepharis macularius. The results indicate that incubation temperature, steroid hormones, and gonads interact in the development of morphological and physiological characters with incubation temperature resulting in the greatest differences in adult phenotype. Incubation temperature did not affect reproductive success directly, but may influence offspring survival in natural situations through effects on adult female body size. Postnatal hormones seem to be more influential in the formation of adult phenotypes than prenatal hormones. These results demonstrate that TSD species can be used to investigate the effects of the physical environment on development in individuals without a predetermined genetic sex and thus provide further insight into the roles of gonadal sex and the embryonic environment in sexual differentiation.

Animals↗
PubMed1995

Making males from females: the effects of aromatase inhibitors on a parthenogenetic species of whiptail lizard.

The parthenogenetic whiptail lizard Cnemidophorus uniparens provides a good model for the study of sex determination and sexual differentiation because genetic variation is minimal and all unmanipulated embryos will develop as females. Thus any deviation from the established course of development can be identified as a treatment effect. Previous work has shown that early prenatal treatment with CGS16949A, a nonsteroidal aromatase inhibitor, causes hatchlings to develop as males. The present study explores more fully the effects of dosage and timing of application of CGS16949A and examines the sex-reversing potential of CGS20267, a new and reputedly more potent aromatase inhibitor. Eggs were treated with a range of dosages of the aromatase inhibitors. Hatchlings that received 1 microgram or more of either inhibitor were all male, while those that received 0.1 microgram or less were all female. No difference in potency between the two compounds was detected. Animals treated with 100 micrograms of CGS16949A on Day 20 of incubation or later were all female, while those treated on Day 5 were all male. Seven sex-reversed male parthenogens have been raised to sexual maturity. The animals appear similar morphologically and behaviorally to males of the sexually reproducing whiptail species. Spermatogenesis and spermiogenesis have been confirmed by histological examination of the testes and by postcopulatory cloacal swabs. Application of aromatase inhibitors has been shown to sex-reverse both avian and reptilian species. In mammals, the male-determining gene of the Y chromosome (SRY) may code for an intrinsic aromatase inhibitor. Studies show the gene's product has a binding domain which recognizes regulatory elements in the promoter of the aromatase gene.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1995

Steroid-induced sex determination at incubation temperatures producing mixed sex ratios in a turtle with TSD.

Previous studies have shown that exogenous steroid hormones can affect sex determination in reptiles with temperature-dependent sex determination (TSD). These studies have also suggested that the sensitivity of TSD to exogenous steroids may vary with incubation temperature. The majority of these studies, however, have utilized incubation temperatures producing all males or all females in the control groups, rather than temperatures which produced mixed sex ratios in control groups. The goals of the current study were to examine the effects of steroids on sex determination in a turtle (Trachemys scripta) at temperatures which produced mixed sex ratios in the control groups. Collectively, the results of single-treatment experiments indicate that at incubation temperatures producing mixed sex ratios in control groups, (1) estradiol-17 beta, tamoxifen, norethindrone, and testosterone all showed a similar "type" of effect (i.e., feminizing) as in previous studies utilizing male-producing temperatures, (2) sex determination has significantly increased sensitivity to estradiol-17 beta in comparison to its effect at temperatures producing all males, and (3) sex determination is sensitive to the masculinizing effects of dihydrotestosterone (DHT) (in previous studies utilizing female-producing temperature DHT did not affect sex determination). Last, a set of double-treatment experiments was performed in which eggs received both estradiol-17 beta and DHT treatments. No significant increases in the production of males were detected. Significant increases in the production of females were detected, but only in the groups receiving the highest dosage of estradiol-17 beta (1.0 micrograms).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1995

The relative effectiveness of androstenedione, testosterone, and estrone, precursors to estradiol, in sex reversal in the red-eared slider (Trachemys scripta), a turtle with temperature-dependent sex determination.

In many turtles the temperature during the middle of incubation determines the gonadal sex of the hatchling. Sex steroid hormones have been implicated in temperature-dependent sex determination in the red-eared slider turtle, Trachemys scripta; androgen is involved in male sex determination and estradiol in female sex determination. Administration of exogenous estradiol and its agonists to eggs incubating at a male-producing temperature can overcome the effect of temperature and result in all-female offspring. Exogenous testosterone will also result in some female hatchlings if administered to eggs incubated at a male-producing temperature, an effect due to the aromatization of testosterone to estradiol. This study demonstrates that in the red-eared slider, androstenedione, the precursor to both testosterone and estradiol, has a similar effect. In addition, both testosterone and androstenedione synergize with incubation temperature to exert a greater effect at intermediate incubation temperatures that normally produce mixed sex ratios, indicating that as with estradiol, androstenedione and testosterone are involved in the final common pathway of sex determination in this species. At the single dosage administered, estrone and estradiol produced all females at a male-producing incubation temperature.

Androstenedione↗
PubMed1995

Seasonal changes and annual variability in daily plasma melatonin in the red-sided garter snake (Thamnophis sirtalis parietalis).

We report seasonal and annual variation in the daily cycle of plasma melatonin levels in male red-sided garter snakes, Thamnophis sirtalis parietalis. In autumn of 1989 and 1990, levels averaged a maximum of 210 pg/ml during scotophase and a minimum of 45 pg/ml during photophase and had a similar diel pattern. Under hibernation conditions (4 degrees, 0:24 L:D), melatonin was undetectable and a diel cycle could not be determined. In Spring 1990, melatonin levels rose rapidly and precipitously within an hour of emergence (while in photophase), peaked at levels significantly higher than those seen in the autumn (approximately 900 pg/ml) and remained significantly high for 24 hr after emergence (though the majority of animals did have decreased levels at the 0400 sample). By the 10th day after emergence, a diel cycle was reestablished and absolute melatonin levels had decreased. The next spring (1991), melatonin again rose within an hour after emergence, while in photophase, but not as high as the previous year. Also unlike the previous year, a diel cycle was observed within 24 hr of emergence. Melatonin levels at emergence were significantly higher than those observed 10 days later. Disruption of a diel rhythm of plasma melatonin (by pinealectomy the previous autumn) inhibits courtship behavior by males the next spring, implying a role for melatonin in the stimulation of sexual behavior. Males in 1991 (with quickly established melatonin cycles) courted much sooner after emergence than did males in 1990. Therefore, the initial day/night difference in melatonin levels at emergence (i.e., establishment of a normal diel cycle) may function in synchronizing and modulating reproductive behavior in male red-sided garter snakes.

Animals↗
PubMed1995

Effect of hormonal manipulation on sociosexual behavior in adult female leopard geckos (Eublepharis macularius), a species with temperature-dependent sex determination.

Aggressive and sexual behavior in the adult leopard gecko (Eublepharis macularius), a species with temperature-dependent sex determination (TSD), is influenced by the temperature experienced as an egg, as well as by prenatal and perinatal hormones. This study focused on the effects of hormonal manipulation of adult female leopard geckos from different incubation temperatures. Following ovariectomy, females from both all-female (26 degrees C) and male-biased (32.5 degrees C) incubation temperatures exhibited a significant decrease in high-posture (HP) aggression toward male and female stimulus animals. Testosterone treatment attenuated this decrease in HP aggression toward female but not toward male stimulus animals. Ovariectomy also resulted in a loss in attractiveness in both groups of females. Following treatment with testosterone, over 50% of the females were attacked by male stimulus animals, suggesting a change in the pheromonal cues normally secreted by females. Unmanipulated females never exhibit tail vibrations, a male-typical courtship behavior. However, following ovariectomy with testosterone treatment, half of the females from both incubation temperatures exhibited this behavior, indicating an activational effect of testosterone. An effect of incubation temperature on aggression was evident with females from the male-biased incubation temperature exhibiting a greater likelihood of aggression compared to females from the all-female incubation temperature. This effect continued to be detected after hormone manipulation. Ovariectomized females from the all-female incubation temperature were less aggressive even with testosterone treatment toward males, whereas females from the male-biased incubation temperature showed no significant decline in aggression following testosterone treatment, suggesting that individuals from different incubation temperatures may have different sensitivities to hormones.

Aggression↗
PubMed1995

Progesterone modulation of androgen-dependent sexual behavior in male rats.

The present study examines the effects of physiological levels of progesterone (P) on copulatory behavior in sexually naive male rats. Two weeks after gonadectomy males were implanted with either empty Silastic capsules (BL) or Silastic capsules containing testosterone (T), P, or both (P+T). When tested with an estrous female, all of the gonadally intact males (intact) and none of the BL controls exhibited mounting/intromission behaviors. Mounting was observed in 75% of the T-alone males. More than half (64%) of the P-alone males and 100% P+T males exhibited mounting. In most cases, mounting was followed by intromission responses. Subsequently, intact and gonadectomized males received daily injections of the P antagonist RU486 along with hormone treatment. After receiving RU486, only 63% of the intact males and 71% of the T-alone males mounted successfully. The facilitatory effects of P on copulatory behavior were completely abolished by RU486 treatment. The present studies provide the first evidence in mammals suggesting that P-dependent mechanisms influence neurochemical pathways involved in copulation.

Androgens↗
PubMed1995

Reptilian sex steroid receptors: amplification, sequence and expression analysis.

Sex steroid hormones secreted by the gonads play a central role in the reproduction of all vertebrates. In addition to direct effects on gametogenesis, sex steroid hormones are important in sexual development, brain organization, and sexual behavior. The actions of sex steroid hormones are mediated primarily by ligand-dependent transcription factors, or receptors which bind to specific sequences of the DNA and alter the transcription rates of nearby genes. We have used the polymerase chain reaction to amplify cDNA fragments of the estrogen receptor, progesterone receptor and androgen receptor from the unisexual whiptail lizard, Cnemidophorus uniparens. The lizard steroid hormone receptors share a high degree of sequence homology to the steroid hormone receptors of other vertebrates. Ribonuclease protection assays demonstrate that both estrogen receptor mRNA and progesterone receptor mRNA are increased in the oviduct during vitellogenesis and after estrogen treatment. This report demonstrates the utility of the polymerase chain reaction to generate species specific probes for comparative molecular studies and provides the first report of cDNA sequences for reptilian steroid hormone receptors.

Amino Acid Sequence↗
PubMed1995

Regulation of estrogen receptor and progesterone receptor messenger ribonucleic acid by estrogen in the brain of the whiptail lizard (Cnemidophorus uniparens).

Receptive behavior in females vertebrates is controlled by hormones, principally estrogen, secreted by the ovary. Estrogen influences behavior by interacting with a specific estrogen binding protein, or receptor, located in target cells in certain hypothalamic nuclei. To better understand the molecular mechanisms involved in the control of receptive behavior in whiptail lizards, we investigated the effects of exogenous estrogen on the regulation of estrogen receptor and progesterone receptor expression in several regions of the brains of Cnemidophorus uniparens. First we determined a dosage of 17 beta-estradiol 3-benzoate (0.5 micrograms) which reliably induced receptive behavior in ovariectomized C. uniparens. Then using in situ hybridization, we examined the effects of that dosage on the expression of estrogen receptor and progesterone receptor mRNA in the brain 24 h after injection. Estrogen treatment resulted in a significant up-regulation of estrogen receptor mRNA expression in the ventromedial nucleus of the hypothalamus and torus semicircularis, down-regulation of estrogen receptor mRNA expression in the lateral septum, and no change in the periventricular nuclei of the hypothalamus, the periventricular nucleus of the preoptic area, and the dorsal hypothalamus. The same dosage resulted in increased progesterone receptor mRNA expression in the ventromedial nucleus of the hypothalamus and the periventricular nucleus of the preoptic area; no significant changes in progesterone receptor mRNA expression were observed in the periventricular nuclei of the hypothalamus or the torus semicircularis, although the differences in progesterone receptor expression in the torus semicircularis approached statistical significance.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1995

Species differences in estrogen receptor and progesterone receptor-mRNA expression in the brain of sexual and unisexual whiptail lizards.

Circulating concentrations of gonadal steroid hormones and reproductive behavior in female vertebrates vary as a function of ovarian state. Steroids secreted by the ovary, specifically estrogen and progesterone, influence the expression of behaviors associated with reproduction by intracellular sex steroid receptors located in specific regions of the brain. Using in situ hybridization, we analyzed estrogen receptor and progesterone receptor messenger RNA expression in several brain regions of ovariectomized, vitellogenic, and postovulatory individuals from two species of whiptail lizards (Cnemidophorus uniparens and C. inornatus). Although these species are genetically very similar, they differ in two aspects of their reproductive biology: (i) the unisexual C. uniparens alternate between expressing female-typical and male-like pseudosexual behaviors while female C. inornatus normally express only female receptive behavior, and (ii) circulating estradiol concentrations in reproductively active female C. uniparens are approximately five-fold lower than in reproductively active female C. inornatus. We found that the regulation of sex steroid receptor gene expression was region specific, with receptor-mRNA expression being increased, unchanged, or decreased during vitellogenesis depending on the area. Furthermore, several species differences in the amount of sex steroid receptor-mRNA were found that may be relevant to the species differences in circulating estrogen concentrations and sexual behavior.

Animals↗
PubMed1995

Species differences in behavioral and neural sensitivity to estrogen in whiptail lizards: correlation with hormone receptor messenger ribonucleic acid expression.

Cnemidophorus uniparens is a unisexual species of whiptail lizard of hybrid origin whereas C. inornatus is a sexual species and the maternal ancestor of C. uniparens. Together they represent an excellent model system for investigating the evolution of hormone-brain-behavior relationships. Normal circulating estradiol (E) concentrations in C. uniparens are approximately 5-fold lower than those of female C. inornatus in a similar reproductive state. Experiments were performed to determine whether (i) C. uniparens is more sensitive to E, and (ii) whether the difference in sensitivity is correlated with differences in estrogen receptor (ER)-mRNA expression in the brain. Dose-response curves reveal that ovariectomized C. uniparens are more responsive than ovariectomized C. inornatus to exogenous estradiol 17 beta-benzoate (EB). EB is more effective in C. uniparens at inducing receptive behavior and progesterone receptor (PR) gene expression in the ventromedial nucleus of the hypothalamus (VMH). In situ hybridization analysis of ER-mRNA expression revealed no species differences in ER-mRNA content in the VMH of ovariectomized animals. Treatment of ovariectomized animals with EB resulted in a greater induction of ER-mRNA expression in the VMH of C. uniparens compared to C. inornatus. These results indicate that the differences in behavioral sensitivity to E lie in the estrogen target neurons in the brain region controlling receptive behavior, the VMH, and that the difference in sensitivity cannot be explained by species differences in the basal rate of ER gene expression.

Animals↗
PubMed1995

Sex differences in estrogen and progesterone receptor messenger ribonucleic acid regulation in the brain of little striped whiptail lizards.

Sex differences in the regulation of steroid hormone receptors in brain areas controlling female- and male-typical sexual behavior may be important in determining sex differences in the display of these behaviors. This study examined sex differences in estrogenic effects on the relative abundance of messenger RNA for estrogen receptor (ER) and progesterone receptor (PR) in discrete brain areas of whiptail lizards, Cnemidophorus inornatus, by in situ hybridization with radiolabeled riboprobes. Gonadectomized females and males received an estradiol benzoate (EB) injection (0.5 microgram) which effectively induces receptive behavior in females; controls received vehicle alone. Sex and regional differences in estrogenic effects on ER- and PR-mRNA abundance were found. Females responded to EB treatment with increases in ER- and PR-mRNA relative abundance in the ventromedial nucleus of the hypothalamus (VMH). Males had similar relative mRNA abundances to females in gonadectomized controls, but did not exhibit increases with EB treatment. EB treatment increased ER-mRNA abundance in the dorsal hypothalamus of females, but not males. ER-mRNA decreases in the lateral septum and PR-mRNA increases in the posterior hypothalamus with hormone treatment were also found, but did not differ by sex. Neither sex nor treatment effects were definitively shown for ER- or PR-mRNA abundance in the anterior hypothalamus-preoptic area. The VMH controls female-typical receptive behavior in this species. Sex differences in the response to estrogen in this nucleus may therefore underlie sex differences in the display of receptive behavior.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗