The present experiments sought to identify brain regions in which implants of an antiestrogen would antagonize the ability of a systemic estradiol treatment to activate sexual behavior in female rats. In experiment 1, ovariectomized female rats were implanted subcutaneously with 5-mm Silastic capsules containing a 5% concentration of estradiol and injected with 500 micrograms progesterone 2 days later, 4-5 h before testing for sexual behavior. Bilateral intracranial implants of 1% crystalline concentrations of the high-affinity antiestrogens monohydroxytamoxifen (TAM) or keoxifene placed into the ventromedial nucleus of the hypothalamus (VM) 24 h prior to estradiol treatment significantly reduced lordosis responsivity compared with control females receiving empty cannulae. Implants of 1% TAM into the medial preoptic area or medial amygdala 24 h prior to estradiol that no significant effect on lordosis. Similarly, implants of 1% TAM into the VM 12 h after estradiol had no effect on lordosis. In experiment 2, lordosis was activated by subcutaneous implants of Silastic capsules containing 1% estradiol plus 500 micrograms progesterone. In this experiment, implants of 1% TAM into the VM 24 h prior to estradiol significantly reduced lordosis only if both cannulae tips were in, or adjacent to, the VM. Females receiving intracranial 1% TAM, but whose cannulae (even unilaterally) were outside the VM, had levels of lordosis equivalent to those of control females. Increasing the concentration of intracranial TAM to 10% virtually eliminated lordosis in females with bilateral implants in the VM, whereas females receiving intracranial 10% TAM in the region of, but outside, the VM showed no evidence of a lordosis deficit.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1987

Progestin receptors with and without estrogen induction in male and female hamster brain.

Progestin receptors have been detected in the brain and pituitary of gonadectomized hamsters using the synthetic ligand (3H)R5020. The number of receptors found in males and females could be dramatically increased by priming the animals with estradiol benzoate 48 h before the binding was measured. Total progestin-binding activity in the cortex, hypothalamus-preoptic area and pituitary of both sexes was the same when high doses of estrogen (30 micrograms per animal) were used for induction, but the inducibility of the male receptors was found to decline with time after gonadectomy. These findings are discussed in relation to the effects of progesterone on aggressive and sexual behaviors in gonadectomized hamsters.

Animals↗

PubMed1986

Identification of medial preoptic neurons that concentrate estradiol and project to the midbrain in the rat.

Fluorescent dye retrograde tracing was combined with steroid hormone autoradiography to study the midbrain projections of the estrogen-concentrating neurons in the preoptic region of the rat brain. Microinjections of the dyes DAPI, true blue, or a mixture of DAPI and primuline were made into the ventral tegmental area and into the midbrain central gray of ovariectomized, adrenalectomized 2-3-month-old female rats; 3 or 4 days later these animals were injected with [3H]estradiol; the brains were then processed for autoradiography. After exposures of from 3 to 12 months, the autoradiograms were developed and examined for reduced silver grains under cell nuclei (indicating binding of [3H]estradiol) and retrogradely transported fluorescent dye in the cytoplasm (indicating an efferent projection to the midbrain). Numerous [3H]estradiol-concentrating neurons in the medial preoptic region were found to send their axons to the medial midbrain. The largest numbers of estrogen target neurons that were afferent to the ventral tegmental area and to the midbrain central gray were found in the medial preoptic nucleus, in the surrounding medial preoptic area, and in the ventral bed nucleus of the stria terminalis. Double-labeled neurons were also identified in the preoptic suprachiasmatic area, in the lateral preoptic area, and in the rostral anterior hypothalamic area. Thus, a subset of the gonadal steroid target cells of the preoptic region have long projections to the medial midbrain, and a subset of the medial preoptic neurons that project to the ventral tegmental area and to the midbrain central gray concentrate estrogen. Behaviors (for example, maternal behavior, male copulatory behavior, and wheel-running) that are regulated by estrogen action in the medial preoptic region may be controlled by the direct estrogen-sensitive pathway to the medial midbrain revealed in this study.

Animals↗

PubMed1986

Correlations between EEG state and spontaneous and evoked axial muscle EMG.

The occurrence of changes in the spontaneous EMG activity of the axial muscle lateral longissimus (LL) is highly correlated (P less than 0.001) with the occurrence of changes in EEG state in urethane-anesthetized rats. There is also a highly significant correlation (P less than 0.001) between the desynchronized EEG state and pudendal nerve evoked multi-unit EMG activity in LL. It is suggested that reticular activation upon pudendal nerve stimulation could facilitate subsequent lumbar axial motoneuron responses to this stimulation.

Animals↗

PubMed1986

Gene expression in hypothalamic neurons: luteinizing hormone releasing hormone.

In situ hybridization allows the detection and measurement of specific messenger RNAs in individual hypothalamic neurons, and has shown, among magnocellular neurons, not only which cells express the genes for oxytocin and vasopressin but also how they change with physiological stimulation. With this technique, neurons expressing a gene for luteinizing hormone releasing hormone-like messenger RNA have been discovered in the preoptic area and diagonal bands of the rat forebrain. Seven days of estrogen treatment of ovariectomized female rats increases the LHRH-like messenger RNA in this neuronal system.

Animals↗

PubMed1986

Quantitative autoradiographic analysis of estradiol retention by cells in the preoptic area, hypothalamus and amygdala.

These experiments were done to compare quantitatively, on a cell-by-cell basis, estradiol retention by cells in the medial preoptic area, arcuate nucleus, ventrolateral subdivision of the ventromedial nucleus, and the caudal half of the medial nucleus of the amygdala. The steroid autoradiograms were prepared from 2 mu sections of brains from ovariectomized, adrenalectomized adult female rats that had been infused intravenously with [3H] estradiol (E2) in a regimen which kept circulating hormone concentration at or above proestrus levels for 3-4 h. Even in these brain regions, containing the most dense collections of E2-concentrating cells, a maximum of only 27-61% of the cells concentrated E2. Therefore, in these regions only a particular subset of the cells retain hormone; other cells in the region do not retain hormone. Frequency distribution histograms of the number of grains per cell versus the number of cells in each region showed a wide range in the amount of E2 retained per cell, and no modes among E2-retaining cells. The data followed a distribution markedly different from that predicted by a simple Poisson distribution, confirming that E2-retention does not result from a random, passive process such as diffusion. The overall quantitative characteristics of the frequency distribution histograms were similar across the four brain areas. Therefore, we propose that the different E2-sensitive functions of these brain areas must depend on differences in the neural connectivity or differences in hormone regulated peptide content of the areas.

Amygdala↗

PubMed1986

Effect of preoptic region implants of dilute estradiol on the maternal behavior of ovariectomized, nulliparous rats.

Estrogen implanted directly into the medial preoptic region of pregnant Charles River Sprague-Dawley rats hysterectomized and ovariectomized on Day 16 of gestation mimics the effects of systemic estrogen treatment at this time by reducing the latency to respond to foster pups with maternal behavior (Numan, Rosenblatt, and Komisaruk, 1977). The present report describes the pup-directed responses of ovariectomized, nulliparous Zivic-Miller Sprague-Dawley rats that received bilateral medial preoptic implants of either cholesterol (n = 11) or estradiol diluted 1:10 with cholesterol (n = 11). Two days after treatment these animals were housed with three foster pups: their responsivity to pups and quality of nests built were then assessed, at first hourly and then daily. Rats receiving intracranial estradiol required significantly shorter exposures to pups than did cholesterol-treated animals before initiating carrying and grouping of 3 dispersed pups in a maternal nest during a 15-min test. On other measures, however, the groups did not differ (e.g., proportion grouping pups overnight, time required to complete retrieval of pups to the nest, time required to rebuild a disrupted nest). Animals treated with cholesterol and animals with estradiol implants did not differ in uterine weight at the time of sacrifice, suggesting that estrogen did not leak, even from this well-vascularized implant site, into the circulation. Thus, as in the pregnant animal, the facilitating effects of estrogen on maternal behavior can be mediated through the medial preoptic region; however, the effects were evident only when a test requiring retrieval of several pups within an arbitrarily short interval was given.

Animals↗

PubMed1986

Effect of varying the duration of pre-test cage habituation on oxytocin induction of short-latency maternal behavior.

Intracerebroventricular (ICV) infusion of the peptide hormone oxytocin has been previously reported to induce the performance of short-latency maternal behavior (less than one hour of exposure to foster pups required) in estrogen-treated, ovariectomized virgin rats. Tests for the effect of ICV oxytocin in maternal behavior latency have included transfer of animals from their home cage to a larger test cage one to two hours before oxytocin infusion. The importance of this test feature on peptide-induced short-latency maternal behavior was evaluated by varying the duration of the pre-test cage habituation. The responses of ovariectomized, estrogen-primed Zivic-Miller Sprague-Dawley rats housed in the test cages one week, two hours, or 0 hours before oxytocin or saline infusion were compared. It was found that only the rats given two hours of pre-test cage habituation responded to ICV oxytocin treatment with short-latency maternal behavior. This result is discussed with regard to the failure of other investigators to elicit short-latency maternal behavior with oxytocin. Possible neuroendocrine mechanisms for the interaction of degree of environment novelty with oxytocin-induced behavior are considered.

Animals↗

PubMed1986

Vasopressin excites ventromedial hypothalamic glucose-responsive neurons in vitro.

Effects of vasopressin (AVP), oxytocin (OXY), norepinephrine (NE), and glucose on the single-unit activity of hypothalamic ventromedial nucleus (VMN) in tissue slices were studied. While AVP was exclusively excitatory on 58% of the neurons, OXY could be excitatory or inhibitory and affected only 42% of the neurons. There was no correlation between the responses to these two peptides. Each of these two peptides could desensitize neuronal response to itself, but did not cross-desensitize responses to each other. These results indicate that AVP and OXY do not act on the same population of VMN neurons through the same cellular mechanism. Furthermore, only the responses to AVP were correlated to responses to glucose and NE, two agents relevant to central regulation of feeding. This correlation with responses to feeding-relevant agents and the exclusively excitatory action on the VMN, which is involved in the regulation of feeding, suggest that AVP can play a role in the regulation of feeding, particularly the feeding induced by the injection of NE into the paraventricular nucleus, that is known to alter AVP release.

Animals↗

PubMed1986

In situ hybridization for the study of gene expression in the brain.

Explore the source record for details and available documents.

Animals↗

PubMed1986

Reversible disruption of lordosis via midbrain infusions of procaine and tetrodotoxin.

Behavioral effects of bilateral intracranial infusions of tetrodotoxin (1, 3.3 or 10 ng/rat), 50% procaine (2 microliters/rat) or phosphate-buffered saline (PBS-2 microliters/rat) into the dorsal midbrain of conscious, lightly-restrained female rats were evaluated. High levels of lordotic responsiveness were induced in ovariectomized animals treated with estradiol (E2) capsules or subcutaneous injections of estradiol benzoate (EB) followed by progesterone (P). The effect of each of the 3 infusates on lordosis was determined using manual stimulation and lordosis quotient determinations. In addition, the vocalization by an animal during lordosis measurements, paw withdrawal to pinch, righting reflex latency and recognition of a platform edge were also monitored. Within 2 minutes following procaine or tetrodotoxin (TTX) infusions in E2 implanted rats, lordotic responsiveness declined sharply. Whereas procaine-treated animals returned to control levels of responsiveness within 20 minutes, TTX infusions induced a more prolonged depression of lordosis lasting up to 8 hours. Infusions of PBS had no effect on any of the behaviors. In a separate group of animals treated with either E2 or EB + P and infused with 10 ng TTX the time course of the decline in lordotic responsiveness was identical for both steroid treatments. Paw withdrawal was unaffected by TTX while all other measured behaviors were disrupted along the same time course as lordosis. Collectively the above results implicate the requirement of sodium-dependent neuronal activity within dorsal midbrain for the maintenance of the lordosis reflex, along with other behavioral responses influenced by this brain region.

Animals↗

PubMed1986

In situ hybridization detection of estradiol-induced changes in ribosomal RNA levels in rat brain.

In this study, quantitative assessment of estradiol (E2)-induced changes in levels of ribosomal RNA within brain regions concentrating the hormone was accomplished by in situ hybridization with nick-translated tritiated ribosomal DNA probes and use of a computer-based image analysis system. Ovariectomized rats were either implanted with estradiol capsules for 6 h, 24 h, or 15 days, or sham-implanted under the same time course to serve as controls. The mean number of grains, somal area, and grain density of neurons within three E2-concentrating brain regions, the ventrolateral portion of the ventromedial and the arcuate nuclei of the hypothalamus (VL-VMN and ARC, respectively) and the corticomedial nucleus of the amygdala (AMY) were determined. In the VL-VMN and ARC, levels of rRNA were significantly increased after 6 h of E2 treatment (70% and 30%, respectively) and after 24 h of E2 treatment (60% and 62%, respectively). However, these effects on rRNA levels in VL-VMN and ARC were not observed after prolonged exposure of 15 days to the hormone. Neuronal hypertrophy was present only after 24 h of E2 treatment in the VL-VMN and ARC (32% and 14%, respectively). No changes were found in the AMY. As an additional internal control, measurements were also collected from the dorsomedial portion of the VMN (DM-VMN), a region with few E2-concentrating neurons. No changes in any of the parameters were found in DM-VMN at any time after exposure to the hormone. By extending the in situ hybridization technique to the quantitative level, these findings demonstrate differential estrogenic regulation of a known gene product, rRNA, in rat brain that is temporally and regionally specific.

Animals↗

PubMed1986

Electrophysiological test of an amphiphilic beta-structure in LHRH action.

Micropipettes were used to record electrical activity from single neurons in hypothalamic tissue slices, in the hypothalamic arcuate nucleus (ARC), and in the periventricular and suprachiasmatic preoptic nuclei (POA). Responses were measured following in vitro application of luteinizing hormone releasing hormone (LHRH), and two analogues: LHRH models 1 and 3. Model 1 (pyroGlu-His-Trp-Ser-Phe-Thr-Ile-Lys-Ile-ThrNH2) had amino acid substitutions in residues 5-10 designed to form an amphiphilic beta-strand structure. Model 3 (pyroGlu-His-Trp-Ser-Phe-Gly-Ile-Lys-Pro-SerNH2) was also designed to possess amphiphilic characteristics, but also more closely to resemble the native peptide. Electrical recording results showed that LHRH was able both to excite or inhibit different hypothalamic neurons, and that it was more effective in the preoptic area than in the arcuate nucleus. Responses to LHRH model 3 were strongly correlated with responses to LHRH, in their occurrence and their direction, for both of the brain regions studied. Moreover, LHRH models 1 and 3 also retained the neuromodulatory effects of LHRH on cellular responses to norepinephrine and serotonin. Thus, LHRH analogues, designed to possess an amphiphilic beta-structure, preserved some of the properties of LHRH when tested electrophysiologically in the central nervous system.

Amino Acid Sequence↗