Anti-Müllerian hormone may regulate the number of calbindin-positive neurons in the sexually dimorphic nucleus of the preoptic area of male mice

Background

The male brain is putatively organised early in development by testosterone, with the sexually dimorphic nucleus of the medial preoptic area (SDN) a main exemplifier of this. However, pubescent neurogenesis occurs in the rat SDN, and the immature testes secrete anti-Müllerian hormone (AMH) as well as testosterone. We have therefore re-examined the development of the murine SDN to determine whether it is influenced by AMH and/or whether the number of calbindin-positive (calbindin^+ve) neurons in it changes after pre-pubescent development.

Methods

In mice, the SDN nucleus is defined by calbindin^+ve neurons (CALB-SDN). The number and size of the neurons in the CALB-SDN of male and female AMH null mutant (Amh ^-/- ) mice and their wild-type littermates (Amh ^+/+ ) were studied using stereological techniques. Groups of mice were examined immediately before the onset of puberty (20 days postnatal) and at adulthood (129–147 days old).

Results

The wild-type pre-pubertal male mice had 47% more calbindin^+ve neurons in the CALB-SDN than their female wild-type littermates. This sex difference was entirely absent in Amh ^-/- mice. In adults, the extent of sexual dimorphism almost doubled due to a net reduction in the number and size of calbindin^+ve neurons in females and a net increase in neuron number in males. These changes occurred to a similar extent in the Amh ^-/- and Amh ^+/+ mice. Consequently, the number of calbindin^+ve neurons in Amh ^-/- adult male mice was intermediate between Amh ^+/+ males and Amh ^+/+ females. The sex difference in the size of the neurons was predominantly generated by a female-specific atrophy after 20 days, independent of AMH.

Conclusions

The establishment of dimorphic cell number in the CALB-SDN of mice is biphasic, with each phase being subject to different regulation. The second phase of dimorphism is not dependent on the first phase having occurred as it was present in the Amh ^-/- male mice that have female-like numbers of calbindin^+ve neurons at 20 days. These observations extend emerging evidence that the organisation of highly dimorphic neuronal networks changes during puberty or afterwards. They also raise the possibility that cellular events attributed to the imprinting effects of testosterone are mediated by AMH.

     

Did sexually dimorphic dorsal coloration evolve by a pre-existing bias in males in the lizard Sceloporus minor?

Theory and empirical evidence indicate that male secondary sex traits can evolve by co-option of pre-existing biases in females. However, relatively few studies have explored whether male pre-existing biases could drive the evolution of traits important in male contests. Male spiny lizards (Sceloporus) are characterized by the expression of sexually dimorphic blue throat and abdominal patches. These features are revealed to conspecifics during social interactions, and variation in ventral color can predict the outcome of male contests in some species of spiny lizards. In Sceloporus minor, males in some populations also express bright blue color on the dorsal surfaces. Given the significance of blue color in intrasexual signaling in other species of Sceloporus, blue dorsal color may have evolved in S. minor by co-option of a male sensory bias for the color blue. We tested this hypothesis in a population that exhibits an ancestral phenotype for male dorsal color (brown/orange), and lacks males with bright blue dorsal coloration. Resident territorial males were presented with one of three types of intruder males manipulated in dorsal color by painting. Orange males mimicked the ancestral dorsal phenotype found at the study site; blue males resembled those from a population with the derived (blue) form of this trait; and green males represented a novel stimulus control. If blue dorsal color evolved in S. minor in part due to co-option of a male sensory bias, we predicted that resident males would exhibit either increased or decreased levels of aggression to blue intruders relative to controls. We found no difference in resident aggressive behavior across all treatments, thus failing to support the predictions of a pre-existing bias. We discuss these findings in the context of social behavior in Sceloporus, and propose directions for further study in this species.     

Is the lateral angle of the internal acoustic canal sexually dimorphic in non-adults? An investigation by routine cranial magnetic resonance imaging

The lateral angle of the internal acoustic canal is one of the measurements of petrous bone that has been previously studied for sex estimation, mostly in adults. We aimed at evaluating the effects of age, side, and sex on the lateral angle of the internal acoustic canal in pediatric patients.Pediatric routine cranial MRI studies were retrospectively investigated for this study. The lateral angle was measured on T2-weighted axial images when the anterior and posterior lips of the meatus and the cochlea were clearly visible on the same image. The data were evaluated for age, side and sex-related changes. Although 552 temporal bones from 273 patients were inspected, due to exclusion criteria lateral angle could be satisfactorily measured only in 101 temporal bones from 58 patients. The measurements did not differ significantly between sexes. An age-related, statistically significant decrease was observed for the entire pediatric sample studied, as well as for the males, but not for females. The measurements did not differ from side to side.The significant age-related decrease in lateral angle in male pediatric patients that was not detected in female counterparts may be the reflection of a sex-related difference in temporal bone development during childhood. Routine cranial MRI data may help investigators study age and sex-related changes in lateral angle in children.     

Ultrastructural localization of α-galactose-containing glycoconjugates in the rat vomeronasal organ

Binding sites of Griffonia simplicifolia I-B₄ isolectin (GS-I-B₄), which recognizes terminal α-galactose residues of glycoconjugates, were examined in the juxtaluminal region of the rat vomeronasal sensory epithelium and its associated glands of the vomeronasal organ, using a lectin cytochemical technique. Lowicryl K4M-embedded ultra-thin sections, which were treated successively with biotinylated GS-I-B₄ and streptavidin-conjugated 10 nm colloidal gold particles, were observed under a transmission electron microscope. Colloidal gold particles, which reflect the presence of terminal α-galactose-containing glycoconjugates, were present in vomeronasal receptor neurons in the sensory epithelium and secretory granules of acinar cells of associated glands of the epithelium. Quantitative analysis demonstrated that the density of colloidal gold particles associated with sensory cell microvilli that projected from dendritic endings of vomeronasal neurons was considerably higher than that of microvilli that projected from neighboring sustentacular cells. The same was true for the apical cytoplasms of these cells just below the microvilli. These results suggest that of the sensory microvilli and dendritic endings contained a much larger amount of the α-galactose-containing glycoconjugates, compared with those in sustentacular microvilli. Further, biochemical analyses demonstrated several vomeronasal organ-specific glycoproteins with terminal α-galactose.     

Gαo and Gαq)regulate the expression of daf-7, a TGFβ-like gene, in Caenorhabditis elegans

Caenorhabditis elegans enter an alternate developmental stage called dauer in unfavorable conditions such as starvation, overcrowding, or high temperature. Several evolutionarily conserved signaling pathways control dauer formation. DAF-7/TGFβ and serotonin, important ligands in these signaling pathways, affect not only dauer formation, but also the expression of one another. The heterotrimeric G proteins GOA-1 (Gα(o)) and EGL-30 (Gα(q)) mediate serotonin signaling as well as serotonin biosynthesis in C. elegans. It is not known whether GOA-1 or EGL-30 also affect dauer formation and/or daf-7 expression, which are both modulated in part by serotonin. The purpose of this study is to better understand the relationship between proteins important for neuronal signaling and developmental plasticity in both C. elegans and humans. Using promoter-GFP transgenic worms, it was determined that both goa-1 and egl-30 regulate daf-7 expression during larval development. In addition, the normal daf-7 response to high temperature or starvation was altered in goa-1 and egl-30 mutants. Despite the effect of goa-1 and egl-30 mutations on daf-7 expression in various environmental conditions, there was no effect of the mutations on dauer formation. This paper provides evidence that while goa-1 and egl-30 are important for normal daf-7 expression, mutations in these genes are not sufficient to disrupt dauer formation.     

Does predation result in adult sex ratio skew in a sexually dimorphic insect genus?

Theory proposes that sexually dimorphic, polygynous species are at particularly high risk of sex-biased predation, because conspicuous males are more often preyed upon compared to females. We tested the effects of predation on population sex ratio in a highly sexually dimorphic insect genus (Hemideina). In addition, introduction of a suite of novel mammalian predators to New Zealand during the last 800 years is likely to have modified selection pressures on native tree weta. We predicted that the balance between natural and sexual selection would be disrupted by the new predator species. We expected to see a sex ratio skew resulting from higher mortality in males with expensive secondary sexual weaponry; combat occurs outside refuge cavities between male tree weta. We took a meta-analytic approach using generalized linear mixed models to compare sex ratio variation in 58 populations for six of the seven species in Hemideina. We investigated adult sex ratio across these populations to determine how much variation in sex ratio can be attributed to sex-biased predation in populations with either low or high number of invasive mammalian predators. Surprisingly, we did not detect any significant deviation from 1 : 1 parity for adult sex ratio and found little difference between populations or species. We conclude that there is little evidence of sex-biased predation by either native or mammalian predators and observed sex ratio skew in individual populations of tree weta is probably an artefact of sampling error. We argue that sex-biased predation may be less prevalent in sexually dimorphic species than previously suspected and emphasize the usefulness of a meta-analytic approach to robustly analyse disparate and heterogeneous data.     

Electroencephalographic signals synchronize with behaviors and are sexually dimorphic during the light–dark cycle in reproductive frogs

Male frogs behave differently from females during the breeding season, particularly with respect to courtship displays and in response to mating signals. In search of physiological correlates of these differences, the present study measured changes in baseline electroencephalogram (EEG) power output within four frequency bands in the telencephalon and mesencephalon, together with changes in locomotor activity as a function of the light–dark cycle in male and female Emei music frogs (Babina daunchina) at the reproductive stage. Previous studies have shown that male vocal activity varies both seasonally and daily in this species and that females use male advertisement calls to locate and select mates. The present results show that both EEG and locomotor activity exhibit highly correlated circadian patterns with peaks around light onset and offset. Importantly, during the reproductive stage, statistically significant sex differences in EEG output across brain regions during the light and dark phases were found indicating that sexual dimorphism exists for EEG activity which may underlie sexually specific information processing and behavioral activities.     

Goα expression in the vomeronasal organ and olfactory bulb of the tammar wallaby

The vomeronasal organ (VNO) detects pheromones via 2 large families of receptors: vomeronasal receptor 1, associated with the protein Giα2, and vomeronasal receptor 2, associated with Goα. We investigated the distribution of Goα in the developing and adult VNO and adult olfactory bulb of a marsupial, the tammar wallaby. Some cells expressed Goα as early as day 5 postpartum, but by day 30, Goα expressing cells were distributed throughout the receptor epithelium of the VNO. In the adult tammar, Goα appeared to be expressed in sensory neurons whose nuclei were mostly basally located in the vomeronasal receptor epithelium. Goα expressing vomeronasal receptor cells led to all areas of the accessory olfactory bulb (AOB). The lack of regionally restricted projection of the vomeronasal receptor cell type 2 in the tammar was similar to the uniform type, with the crucial difference that the uniform type only shows expression of Giα2 and no expression of Goα. The observed Goα staining pattern suggests that the tammar may have a third accessory olfactory type that could be intermediate to the segregated and uniform types already described.     

Voltage-activated current properties of male and female mouse vomeronasal sensory neurons: sexually dichotomous?

The vomeronasal organ, the chemosensory organ of the vomeronasal system, is vital in determining sexual and gender-specific behavior in mice. Here, whole-cell voltage-activated currents of individual mouse vomeronasal sensory neurons of two strains (BALB/c and CBA) were measured and correlated to sex in each strain. The average resting membrane potentials, maximal outward current magnitudes, and kinetics of activation and inactivation, were found to be independent of sex. Maximal inward current magnitudes differed significantly across gender in CBA, whereas they did not significantly differ in male and female BALB/c mice: BALB/c males –347±45 pA (n=51), and females –430±56 pA (n=27); CBA males –308±36 pA (n=56) and females –155±18 pA (n=28). These results suggest that some voltage-activated properties may differ slightly according to gender and to strain.D.M. Dean and A. Mazzatenta contributed equally to this work     

Transposition and Intermingling of Gαi2 and Gαo Afferences into Single Vomeronasal Glomeruli in the Madagascan Lesser Tenrec Echinops telfairi

The vomeronasal system (VNS) mediates pheromonal communication in mammals. From the vomeronasal organ, two populations of sensory neurons, expressing either Gαi2 or Gαo proteins, send projections that end in glomeruli distributed either at the rostral or caudal half of the accessory olfactory bulb (AOB), respectively. Neurons at the AOB contact glomeruli of a single subpopulation. The dichotomic segregation of AOB glomeruli has been described in opossums, rodents and rabbits, while Primates and Laurasiatheres present the Gαi2-pathway only, or none at all (such as apes, some bats and aquatic species). We studied the AOB of the Madagascan lesser tenrec Echinops telfairi (Afrotheria: Afrosoricida) and found that Gαi2 and Gαo proteins are expressed in rostral and caudal glomeruli, respectively. However, the segregation of vomeronasal glomeruli at the AOB is not exclusive, as both pathways contained some glomeruli transposed into the adjoining subdomain. Moreover, some glomeruli seem to contain intermingled afferences from both pathways. Both the transposition and heterogeneity of vomeronasal afferences are features, to our knowledge, never reported before. The organization of AOB glomeruli suggests that synaptic integration might occur at the glomerular layer. Whether intrinsic AOB neurons may make synaptic contact with axon terminals of both subpopulations is an interesting possibility that would expand our understanding about the integration of vomeronasal pathways.     

Mode of interaction of the Gαo subunit of heterotrimeric G proteins with the GoLoco1 motif of Drosophila Pins is determined by guanine nucleotides

Drosophila GoLoco motif-containing protein Pins is unusual in its highly efficient interaction with both GDP- and the GTP-loaded forms of the α-subunit of the heterotrimeric Go protein. We analysed the interactions of Gαo in its two nucleotide forms with GoLoco1–the first of the three GoLoco domains of Pins–and the possible structures of the resulting complexes, through combination of conventional fluorescence and FRET measurements as well as through molecular modelling. Our data suggest that the orientation of the GoLoco1 motif on Gαo significantly differs between the two nucleotide states of the latter. In other words, a rotation of the GoLoco1 peptide in respect with Gαo must accompany the nucleotide exchange in Gαo. The sterical hindrance requiring such a rotation probably contributes to the guanine nucleotide exchange inhibitor activity of GoLoco1 and Pins as a whole. Our data have important implications for the mechanisms of Pins regulation in the process of asymmetric cell divisions.     

How strong is the mutagenicity of recombination in mammals?

It is commonly believed that a high recombination rate such as that in a pseudoautosomal region (PAR) greatly increases the mutation rate because a 170-fold increase was estimated for the mouse PAR region. However, sequencing PAR and non-PAR introns of the Fxy gene in four Mus taxa, we found an increase of only twofold to fivefold. Furthermore, analyses of sequence data from human and orangutan PAR and X-linked regions and from autosomal regions showed a weak effect of recombination on mutation rate (a slope of less than 0.2% per cM/Mb), although a much stronger effect on GC content (1% to 2% per cM/Mb). Because typical recombination rates in mammals are much lower than those in PARs, the mutagenicity of recombination is weak or, at best, moderate, although its effect on GC% is much stronger. In addition, contrary to a previous study, we found no Fxy duplicate in Mus spretus.     

17β-Estradiol is required for the sexually dimorphic effects of repeated binge-pattern alcohol exposure on the HPA axis during adolescence

Alcohol consumption during adolescence has long-term sexually dimorphic effects on anxiety behavior and mood disorders. We have previously shown that repeated binge-pattern alcohol exposure increased the expression of two critical central regulators of stress and anxiety, corticotrophin-releasing hormone (CRH) and arginine vasopressin (AVP), in adolescent male rats. By contrast, there was no effect of alcohol on these same genes in adolescent females. Therefore, we tested the hypothesis that 17β-estradiol (E(2)), the predominant sex steroid hormone in females, prevents alcohol-induced changes in CRH and AVP gene expression in the paraventricular nucleus (PVN) of the hypothalamus. To test this hypothesis, postnatal day (PND) 26 females were ovariectomized and given E(2) replacement or cholesterol as a control. Next, they were given an alcohol exposure paradigm of 1) saline alone, 2) acute (single dose) or 3) a repeated binge-pattern. Our results showed that acute and repeated binge-pattern alcohol treatment increased plasma ACTH and CORT levels in both E(2)- and Ch-treated groups, however habituation to repeated binge-pattern alcohol exposure was evident only in E(2)-treated animals. Further, repeated binge-pattern alcohol exposure significantly decreased CRH and AVP mRNA in Ch-, but not E(2)-treated animals, which was consistent with our previous observations in gonad intact females. We further tested the effects of E(2) and alcohol treatment on the activity of the wild type CRH promoter in a PVN-derived neuronal cell line. Alcohol increased CRH promoter activity in these cells and concomitant treatment with E(2) completely abolished the effect. Together our data suggest that E(2) regulates the reactivity of the HPA axis to a repeated stressor through modulation of the habituation response and further serves to maintain normal steady state mRNA levels of CRH and AVP in the PVN in response to a repeated alcohol stressor.     

Coordinated control of sensitivity by two splice variants of Gαo in retinal ON bipolar cells

The high sensitivity of scotopic vision depends on the efficient retinal processing of single photon responses generated by individual rod photoreceptors. At the first synapse in the mammalian retina, rod outputs are pooled by a rod “ON” bipolar cell, which uses a G-protein signaling cascade to enhance the fidelity of the single photon response under conditions where few rods absorb light. Here we show in mouse rod bipolar cells that both splice variants of the G_o α subunit, Gα_o1 and Gα_o2, mediate light responses under the control of mGluR6 receptors, and their coordinated action is critical for maximizing sensitivity. We found that the light response of rod bipolar cells was primarily mediated by Gα_o1, but the loss of Gα_o2 caused a reduction in the light sensitivity. This reduced sensitivity was not attributable to the reduction in the total number of G_o α subunits, or the altered balance of expression levels between the two splice variants. These results indicate that Gα_o1 and Gα_o2 both mediate a depolarizing light response in rod bipolar cells without occluding each other’s actions, suggesting they might act independently on a common effector. Thus, Gα_o2 plays a role in improving the sensitivity of rod bipolar cells through its action with Gα_o1. The coordinated action of two splice variants of a single Gα may represent a novel mechanism for the fine control of G-protein activity.     

Carnivore mammals in a fragmented landscape in northeast of São Paulo State,Brazil

São Paulo is the most developed state in Brazil and little of its native vegetation remains. In Luiz Antonio and Santa Rita do Passa Quatro municipalities, only small fragments of cerrado (Brazilian savanna) physiognomies (cerradão, cerrado sensu stricto) and of semideciduous forest have been left, surrounded by eucalyptus silviculture and sugar-cane agriculture. However, that vegetation mosaic still shelters large mammals, including several carnivore species. To detect the carnivores present in such a mosaic area (50,000 ha), and to find out how they use the landscape, we recorded them through 21 camera traps and 21 track plots, during 18 months. Species richness, diversity and relative frequency were evaluated according to the habitat. Ten species were recorded, some of them locally threatened to extinction (Puma concolor, Leopardus pardalis, Chrysocyon brachyurus). Species diversity did not significantly differ among fragments, and although most species preferred one or another habitat, the carnivore community as a whole explored all the study area regardless of the vegetation cover; eucalyptus plantations were as used by the carnivores as the native fragments. Therefore, it seems possible to maintain such animals in agricultural landscapes, where some large native fragments are left and the matrix is permeable to native fauna.     

Characterization of a G protein α subunit encoded gene from the dimorphic fungus-Tremella fuciformis

Antonie van Leeuwenhoek - Tremella fuciformis is a dimorphic fungus which can undertake the reversible transition between yeast and pseudohypha forms. G protein α subunit (Gα) carries...     

What limits the velocity of fast-skeletal muscle contraction in mammals?

In rat skeletal muscle the unloaded shortening velocity (Vo) is defined by the myosin isoform expressed in the muscle fibre. In 2001 we suggested that ADP release from actomyosin in solution (controlled by k(-AD)) was of the right size to limit Vo. However, to compare mechanical and solution kinetic data required a series of corrections to compensate for the differences in experimental conditions (0.5 M KCl, 22 degrees C for kinetic assays of myosin, 200 mM ionic strength, 12 degrees C to measure Vo). Here, a method was developed to prepare heavy meromyosin (HMM) from pure myosin isoforms isolated from single muscle fibres and to study k(-AD) (determined from the affinity of the acto-myosin complex for ADP, KAD) and the rate of ATP-induced acto-HMM dissociation (controlled by K1k+2) under the same experimental condition used to measure Vo). In fast-muscle myosin isolated from a wide range of mammalian muscles, k(-AD) was found to be too fast to limit Vo, whereas K1k+2 was of the right magnitude for ATP-induced dissociation of the cross-bridge to limit shortening velocity. The result was unexpected and prompted further experiments using the stopped-flow approach on myosin subfragment-1 (S1) and HMM obtained from bulk preparations of rabbit and rat muscle. These confirmed that the rate of cross-bridge dissociation by ATP limits the velocity of contraction for fast myosin II isoforms at 12 degrees C, while k(-AD) limits the velocity of slow myosin II isoforms. Extrapolating our data to 37 degrees C suggests that at physiological temperature the rate of ADP dissociation may limit Vo for both isoforms.