Estradiol induces sexual behavior in female túngara frogs

Steroid hormones play an important role in regulating vertebrate sexual behavior. In frogs and toads, injections of exogenous gonadotropins, which stimulate steroid hormone production, are often used to induce reproductive behavior, but steroid hormones alone are not always sufficient. To determine which hormonal conditions promote sexual behavior in female túngara frogs, we assessed the effect of hormone manipulation on the probability of phonotaxis behavior toward conspecific calls in post-reproductive females. We injected females with human chorionic gonadotropin (HCG), estradiol, estradiol plus progesterone, saline, or HCG plus fadrozole (an aromatase blocker) and tested their responses to mating calls. We found that injections of HCG, estradiol, and estradiol plus progesterone all increased phonotaxis behavior, whereas injections of saline or HCG plus fadrozole did not. Since injections of estradiol alone were effective at increasing phonotaxis behavior, we concluded that estradiol is sufficient for the expression of phonotaxis behavior. Next, to determine if estradiol-injected females display the same behavioral preferences as naturally breeding females, we compared mating call preferences of naturally breeding females to those of post-reproductive females injected with estradiol. We found that, when injected with estradiol, females show similar call preferences as naturally breeding females, although they were less likely to respond across multiple phonotaxis tests. Overall, our results suggest that estradiol is sufficient for the expression of sexual responses to mating calls in túngara frogs. To our knowledge, ours is the only study to find that estradiol alone is capable of promoting phonotaxis behavior in a frog.     

Diet alters species recognition in juvenile toads

Whether environmental effects during juvenile development can alter the ontogeny of adult mating behaviour remains largely unexplored. We evaluated the effect of diet on the early expression of conspecific recognition in spadefoot toads, Spea bombifrons. We found that juvenile toads display phonotaxis behaviour six weeks post-metamorphosis. However, preference for conspecifics versus heterospecifics emerged later and was diet dependent. Thus, the environment can affect the early development of species recognition in a way that might alter adult behaviour. Evaluating such effects is important for understanding variation in hybridization between species and the nature of species boundaries.     

Anuran mating calling circuits: inhibition by prostaglandin

Male American toads (Bufo americanus) were induced to mating call in response to electronically simulated, conspecific mating calls. The injection of prostaglandin (PG) F2 alpha caused suppression of mating call answering. Neural correlates of mating calling were triggered by electrical stimulation of the anterior preoptic nucleus in the isolated brainstem of male, Northern leopard frogs (Rana p. pipiens). The addition of PGF2 alpha to the bath completely abolished the correlates of mating calling without changing the correlates of pulmonary respiration. The suppression of mating calling shown here, along with the suppression of release signalling described by Diakow and Nemiroff (1981), supports the hypothesis of a close interrelation between the neural circuits of these two calls. The suppression of the neural correlates of mating calling in an isolated preparation shows a central site of action of the PG. The retention of normal correlates of pulmonary respiration, even after suppression of mating calling correlates, suggests that the generation of mating calling patterns involves the extension and pulsing of the expiratory phase of breathing.     

Immunocytochemistry of luteinizing hormone-releasing hormone in brains of breeding eastern narrow-mouthed toads (Gastrophryne carolinensis)

Unlabeled antibody-enzyme immunocytochemistry was used to localize mammalian-like immunoreactive luteinizing hormone-releasing hormone in brains of breeding eastern narrow-mouthed toads (Gastrophryne carolinensis). Intense immunostaining occurred over the infundibular floor and the median eminence, yet staining of few perikarya in the anterior preoptic area was minimal. This pattern of markedly regional immunostaining differs from that reported for other anurans.     

Central control of mating call production and spawning in the tree frog Hyla arborea savignyi (Audouin): Results of electrical stimulation of the brain

Electrical stimulation of the anterior preoptic nucleus elicited the production of mating calls by male Hyla arborea savignyi. Such calls are composed of pulses resembling those of natural mating calls both in their duration and in their sequential arrangement. As a rule, the amplitude of successive pulses within a group increases more rapidly in the electrically evoked calls than in natural calls. The frogs could be brought into a state of readiness to call, which did not lead to actual calling activity, by stimulation in the anterior preoptic, posterior preoptic, and magnocellular nuclei as well as in the hypothalamus. Stimulation of the anterior preoptic nucleus of one female elicited repeated vocalizations, which may be interpretable as mating calls. The posture adopted by the female for spawning can be elicited in males as well as in females by electrical stimulation of the anterior preoptic nucleus.     

Social signals increase monoamine levels in the tegmentum of juvenile Mexican spadefoot toads (Spea multiplicata)

Monoamines are important neuromodulators that respond to social cues and that can, in turn, modify social responses. Yet we know very little about the ontogeny of monoaminergic systems and whether they contribute to the development of social behavior. Anurans are an excellent model for studying the development of social behavior because one of its primary components, phonotaxis, is expressed early in life. To examine the effect of social signals on monoamines early in ontogeny, we presented juvenile Mexican spadefoot toads (Spea multiplicata) with a male mating call or no sound and measured norepinephrine, epinephrine, dopamine, serotonin, and a serotonin metabolite, across the brain using high-pressure liquid chromatography. Our results demonstrate that adult-like monoaminergic systems are in place shortly after metamorphosis. Perhaps more interestingly, we found that mating calls increased the level of monoamines in the juvenile tegmentum, a midbrain region involved in sensory-motor integration and that contributes to brain arousal and attention. We saw no such increase in the auditory midbrain or in forebrain regions. We suggest that changes in monoamine levels in the juvenile tegmentum may reflect the effects of social signals on arousal state and could contribute to context-dependent modulation of social behavior.     

Behavioural patterns, preference, and motivation of female midwife toads during phonotaxis tests

During two-speaker phonotaxis tests performed with female midwife toads (Alytes obstetricans and A. cisternasii), the occurrence of behavioural patterns tentatively considered to be associated with entering amplexus and making a mate choice was noted. The individual repeatability of choice was assessed as a measurement of reliability of female preference, and the individual repeatability of response was assessed as a measurement of the degree of female motivation. The tests were grouped in two categories, spectral and temporal, according to the acoustic variable addressed in the test. Females selecting the alternative chosen less often by the population responded less often in phonotaxis tests and thus could be less motivated. Female repeatability was significant for temporal tests, but not for spectral tests. The observation of behavioural patterns during phonotaxis tests appears to be a useful complement to mate choice studies in anurans. Electronic Publication     

Reproductive stage and history affect the phonotactic preferences of female midwife toads, Alytes muletensis

We investigated whether the reproductive stage or reproductive history of females affects their preferences for male calls that differ in frequency. We repeatedly tested gravid female midwife toads prior to and immediately after mating, and during ovulation. When females were ovulating they were much more consistent in their choice of call than when they were gravid but not ovulating. When females had recently mated they still showed positive phonotaxis, but did not reliably discriminate between alternative calls. This study is the first to test gravid female anurans repeatedly at different stages in their reproductive cycle (ovulating or not; pre- and postmating). It highlights the need to treat with caution the results from population-based preference studies in which individuals are tested only once, and in which variation in reproductive state is not taken into account. Copyright 2000 The Association for the Study of Animal Behaviour.     

Projection from the ventral bed nucleus of the stria terminalis to the retrorubral field in rats and the effects of cells in these areas on mating in male rats versus gerbils

This research identified the rat counterpart of the lateral cell group of the sexually dimorphic area (SDA) found in medial preoptic area (MPOA) gerbil of gerbils. The lateral SDA (lSDA) is critical for mating in male gerbils and contains most of the SDA cells projecting to the retrorubral field (RRF), a projection that is also important for mating. Therefore, to locate the counterpart of the lateral SDA, we traced the inputs to the rat RRF, which were dense in the ventral part of the bed nucleus of the stria terminalis (BST). To determine if the ventral BST or its projection to the RRF affects mating in male rats, we disrupted them bilaterally by placing cell-body lesions bilaterally in the ventral BST or unilaterally there and in the contralateral RRF. We also studied the effects of RRF lesions in both rats and gerbils. Bilateral ventral BST lesions, which left the medial preoptic nucleus intact, produced persistent and severe mating deficits. Disconnecting the ventral BST from the RRF also had long-lasting, but less severe, consequences. RRF lesions produced only temporary mating deficits in rats, but virtually eliminated mating in gerbils. The recovery of mating in rats after RRF, but not ventral BST, lesions, and the intermediate effects of disconnecting these areas from each other suggest that the ventral BST may contain mating-related projection neurons other than those projecting to the RRF or that its RRF-projecting cells send collaterals to another site. In either case, the pedunculopontine tegmental nucleus or raphe nuclei may be involved.     

Response of ERalpha-IR and ERbeta-IR cells in the forebrain of female rats to mating stimuli

Sexual behavior in female rats depends on the action of estradiol on estrogen receptors (ERs) found in particular brain regions. While hormonal regulation of female sexual behavior requires ERalpha, the possible functions of ERbeta remain to be clarified. Mating stimulation has several behavioral and physiological consequences and induces Fos expression in many brain areas involved in the regulation of reproductive behavior and physiology. In addition, some cells in which mating induces Fos expression coexpress ERalpha. To determine whether cells in which Fos is induced by a particular mating stimulus coexpress ERalpha, ERbeta, or both, we used a triple-label immunofluorescent technique to visualize ERalpha-, ERbeta-, and mating-induced Fos-immunoreactivity (Fos-ir) in neurons in which mating stimulation reliably increases Fos expression. Ovariectomized, hormone-primed rats were either unmated, received 15 mounts, or received 15 intromissions. In the rostral medial preoptic area, Fos-ir was induced by mounts alone primarily in cells coexpressing ERalpha-ir, while Fos-ir was induced by intromissions mainly in cells coexpressing both ERalpha-ir and ERbeta-ir (ERalpha/ERbeta-ir). In the dorsal part of the posterodorsal medial amygdala, Fos-ir was induced by intromissions in cells coexpressing ERalpha-ir and ERalpha/ERbeta-ir. However, in the ventral part of the posterodorsal medial amygdala, Fos-ir was induced by intromissions primarily in cells coexpressing only ERbeta-ir. These data suggest that qualitatively different sexual stimuli may be integrated through distinct ER-containing circuits in the rostral medial preoptic area and posterodorsal medial amygdala. The diversity in coexpression of type of ER in cells in different brain areas after various mating stimuli suggests a role for both ERalpha and ERbeta in the integration of hormonal information and information related to mating stimuli.     

Organization of diencephalon of the sturgeons. Preoptic area

Cytoarchitectonics of preoptic area of diencephalon of chondroid ganoid fish (the sturgeons) was studied in serial sections using Nissl and Bilschowski techniques, the latter in Viktorov’s modification). The preoptic area of the hausen, Huso huso L., Kura sturgeon, Acipenser guldenstaedtii persicus n. kurensis Belyaeff, Caspian sturgeon, Acipenser stellatus Pall. and the sturgeon Acipenser nudioventris Lov. was shown to be one of the largest parts of diencephalon. It is located along the rostral excess of the third ventricle and the anterior third of the ventricle thalamic part. Eight nuclei are identified in the preoptic area: (1) anterior microcellular preoptic nucleus, (2) suprachiasmatic nucleus, (3) accessory preoptic nucleus, (4) intrachiasmatic nucleus, (5) magnocellular preoptic nucleus, (6) posterior microcellular preoptic nucleus, (7) caudal preoptic nucleus, (8) entopeduncular nucleus. Besides, microcellular, magnocellular, and gigantocellular parts are identified in the magnocellular preoptic nucleus. The obtained results indicate high differentiation of this area of the sturgeons among Actinopterygii.     

The preoptic area of the domestic fowl

Summary The preoptic area of the domestic fowl (Gallus gallus) was studied by means of the Golgi technique. At least two regions can be recognized: (i) a medial and (ii) a lateral area, clearly distinguishable laterally from the adjacent telencephalic regions. The dendritic organization of the preoptic area is quite uniform. The neurons can be classified as isodendritic elements. The magnocellular elements are few and irregularly scattered mostly in the periventricular grey of the medial preoptic area. Of relevant interest is also the observation of some bipolar and horizontal neurons in the dorsal part of the medial preoptic area, near the anterior commissure.     

SPECIES RECOGNITION AND SEXUAL SELECTION AS A UNITARY PROBLEM IN ANIMAL COMMUNICATION

We investigated patterns of mating call preference and mating call recognition by examining phonotaxis of female túngara frogs, Physalaemus pustulosus, in response to conspecific and heterospecific calls. There are four results: females always prefer conspecific calls; most heterospecific calls do not elicit phonotaxis; some heterospecific calls do elicit phonotaxis and thus are effective mate recognition signals; and females prefer conspecific calls to which a component of a heterospecific call has been added to a normal conspecific call. We use these data to illustrate how concepts of species recognition and sexual selection can be understood in a unitary framework by comparing the distribution of signal traits to female preference functions.     

Acoustic modulation of neural activity in the preoptic area and ventral hypothalamus of the green treefrog (Hyla cinerea)

Summary Responses of neurons in the preoptic area and ventral hypothalamus to conspecific mating calls or white noise bursts were examined in male green treefrogs (Hyla cinerea) during different seasons. In the winter, 34.3% of preoptic neurons and 46.7% of ventral hypothalamic cells demonstrated significant changes in activity level during presentation of a conspecific mating call. In contrast, only 13.3% of preoptic units and 16.7% of ventral hypothalamic cells responded to the white noise. The percentage of preoptic and hypothalamic units responding to the advertisement call did not differ significantly during the summer breeding season. Type I units exhibited a dramatic increase in activity during acoustic stimulation followed by a rapid return to baseline activity levels after stimulus offset. Type II cells showed a robust activity increase during stimulation, but maintained an intermediate activity level after stimulus offset. In the preoptic area, a third response type exhibited suppressed activity during acoustic stimulation. Although seasonal condition did not alter the percentage of acoustically responsive units within either nucleus, the proportion of Type I units in the ventral hypothalamus was greatest during the summer.Abbreviations MC mating call - NS no stimulus - POA preoptic area - VH ventral hypothalamus - WN white noise     

Intergrading between members of the "regularis group" of toads in South Africa

The mating calls of three members of the " regularis group" of toads (Anura: Bufonidae) that are sympatric in part of South Africa are examined. A naturally occurring population of hybrid toads from Pretoria (South Africa) has previously been described as representing an intergrading between Bufo garmani Meek and Bufo rangeri Hewitt. Examination of the mating calls of these toads provides little evidence of intergrading between Bufo rangeri and Bufo garmani , and indicates the involvement of Bufo regularis Reuss in the hybridization. The mating calls of the hybrids are varied, and are structurally intermediate between those of the parental species. Mating call parameters are found to be useful in separating forms that are difficult to distinguish on morphological grounds alone. They are further found to be a sensitive non-morphological means of ascertaining the affinities of naturally occurring hybrid toads.     

Independent control of sexual and scent marking behaviors of male gerbils by cells in or near the medial preoptic area

This research studied the role of the medial preoptic area and adjacent cell populations in androgen control of scent marking and sexual behavior in male gerbils (Meriones unguiculatus). Experiment 1 replicated previous research showing that implants of testosterone propionate in or near the medial preoptic area reinstate marking behavior in castrates. Implant sites near the diagonal band of Broca or in the posterior part of the medial preoptic area, near the anterior hypothalamus, are more effective than other sites. Experiment 2 showed that medial preoptic area lesions permanently impair sexual behavior despite testosterone stimulation. Experiments 2–4 showed that lesions in or near the medial preoptic area can also disrupt scent marking; however, this behavior gradually recovered in many lesioned males, especially if they received testosterone. The data suggest that both scent marking and sexual behavior are controlled by androgens acting on cells in or near the medial preoptic area, but the cell populations involved in these two behaviors are probably not the same.     

Increased estrogen receptor alpha immunoreactivity in the forebrain of sexually satiated rats

Estrogen receptor alpha (ERalpha) participates in the neuroendocrine regulation of male sexual behavior, primarily in brain areas located in the limbic system. Males of many species present a long-term inhibition of sexual behavior after several ejaculations, known as sexual satiety. It has been shown that androgen receptor density is reduced 24 h after a single ejaculation or mating to satiety, in the medial preoptic area, nucleus accumbens and ventromedial hypothalamus. The aim of this study was to analyze if the density of ERalpha was also modified 24 h after a single ejaculation or mating to satiety. Sexual satiety was associated with an increased ERalpha density in the anteromedial bed nucleus of the stria terminalis (BSTMA), ventrolateral septum (LSV), posterodorsal medial amygdala (MePD), medial preoptic area (MPA) and nucleus accumbens core (NAc). A single ejaculation was related to an increase in ERalpha density in the BSTMA and MePD. ERalpha density in the arcuate (Arc) and ventromedial hypothalamic nuclei (VMN), and serum estradiol levels remained unchanged 24 h after one ejaculation or mating to satiety. These data suggest a relationship between sexual activity and an increase in the expression of ERalpha in specific brain areas, independently of estradiol levels in systemic circulation.     

Gonadal steroid receptors colocalize with central nervous system neurons projecting to the rat prostate gland

Mating-induced Fos-immunoreactive (-ir) cells are colocalized with androgen receptors (AR), estrogen receptors (ER), or both in limbic and hypothalamic areas known to mediate male rat mating behavior. A steroid-responsive neural network might govern copulatory behavior in male laboratory rats that is analogous to the network described in female rats that governs the lordosis response. This hypothesized network in males may synchronize and coordinate sexual behavioral responses with physiological responses of the genitals and the internal organs of reproduction. Therefore, the pseudorabies virus (PRV; Bartha strain), a transneuronal, viral retrograde tract tracer, was microinjected into the prostate gland to label this network. After 7 days, brains from infected animals were processed for immunohistochemical labeling of AR, ER, and PRV. The majority of PRV-ir cells exhibited either AR or ER immunoreactivity in the medial preoptic area, median preoptic nucleus, bed nucleus of stria terminalis, hypothalamic paraventricular nucleus, and zona incerta, areas known to play roles in male rat mating behavior. Other structures such as the central tegmental field/subparafascicular nucleus of the thalamus, central nucleus of the amygdala, and medial amygdala, also important in the display of male copulatory behavior, were less reliably labeled. Collectively, a steroid receptor-containing neuronal circuit, largely contained in the diencephalon, was revealed that likely is involved in the autonomic control of the prostate gland and the consummatory aspects of male rat mating behavior.     

Selectivity of the peripheral auditory system of spadefoot toads (Scaphiopus couchi) for sounds of biological significance

Summary Scaphiopus couchi is a primitive anuran whose vocal repertoire consists of a mating call and a release call. The two calls are distinct and differ in trill rate. Reception of airborne sound is achieved by means of a poorly differentiated region of skin on the head which serves as an eardrum.Whereas more modern anurans possessthree distinct types of auditory nerve fibers, spadefoot toads possess onlytwo types: a low-frequency-sensitive group which exhibits tone-on-tone inhibition and a high-frequency-sensitive group which is not inhibitable. The sharpness of frequency tuning of primary fibers in each group is comparable to more advanced vertebrate species. While the response properties of auditory fibers in the high-frequency-sensitive group are well matched to the spectral and temporal features in the spadefoot's mating call and release call, the low-frequency-sensitive fibers do not respond to these calls. Instead they may be involved in detection of bodily transmitted sounds during clasping, as well as other low-frequency sounds in the environment. The two groups of auditory fibers probably derive from separate auditory organs within the inner ear. Thresholds of auditory nerve fibers in spadefoot toads are relatively poorer than in more advanced anurans, which likely is due to their less developed eardrum. The role of tone-on-tone inhibition in the peripheral auditory system is questioned with regard to its significance in processing sounds of biological value.We wish to dedicate this paper to Jasper J. Loftus-Hills who was killed in a tragic accident near Austin, Texas on June 11, 1974. His post-doctoral appointment in our laboratory and his assistance in collecting spadefoot toads in the field recall fond memories.We also wish to thank R. Sage for helping us collect animals and W. F. Blair for supplying tape recordings ofScaphiopus mating calls. The assistance of J. Paton in photographing the animal in Fig. 1 is gratefully appreciated. This research was supported by the U.S. Public Health Service (NIH Research Grant NS-09244) and the National Science Foundation (Grant GB-18836); travel expenses involved in collecting animals were supported by a Cornell University Research Grant.