Sexual differentiation in three unconventional mammals: spotted hyenas, elephants and tammar wallabies

The present review explores sexual differentiation in three non-conventional species: the spotted hyena, the elephant and the tammar wallaby, selected because of the natural challenges they present for contemporary understanding of sexual differentiation. According to the prevailing view of mammalian sexual differentiation, originally proposed by Alfred Jost, secretion of androgen and anti-Mullerian hormone (AMH) by the fetal testes during critical stages of development accounts for the full range of sexually dimorphic urogenital traits observed at birth. Jost's concept was subsequently expanded to encompass sexual differentiation of the brain and behavior. Although the central focus of this review involves urogenital development, we assume that the novel mechanisms described in this article have potentially significant implications for sexual differentiation of brain and behavior, a transposition with precedent in the history of this field. Contrary to the "specific" requirements of Jost's formulation, female spotted hyenas and elephants initially develop male-type external genitalia prior to gonadal differentiation. In addition, the administration of anti-androgens to pregnant female spotted hyenas does not prevent the formation of a scrotum, pseudoscrotum, penis or penile clitoris in the offspring of treated females, although it is not yet clear whether the creation of masculine genitalia involves other steroids or whether there is a genetic mechanism bypassing a hormonal mediator. Wallabies, where sexual differentiation occurs in the pouch after birth, provide the most conclusive evidence for direct genetic control of sexual dimorphism, with the scrotum developing only in males and the pouch and mammary glands only in females, before differentiation of the gonads. The development of the pouch and mammary gland in females and the scrotum in males is controlled by genes on the X chromosome. In keeping with the "expanded" version of Jost's formulation, secretion of androgens by the fetal testes provides the best current account of a broad array of sex differences in reproductive morphology and endocrinology of the spotted hyena, and androgens are essential for development of the prostate and penis of the wallaby. But the essential circulating androgen in the male wallaby is 5alpha androstanediol, locally converted in target tissues to DHT, while in the pregnant female hyena, androstenedione, secreted by the maternal ovary, is converted by the placenta to testosterone (and estradiol) and transferred to the developing fetus. Testicular testosterone certainly seems to be responsible for the behavioral phenomenon of musth in male elephants. Both spotted hyenas and elephants display matrilineal social organization, and, in both species, female genital morphology requires feminine cooperation for successful copulation. We conclude that not all aspects of sexual differentiation have been delegated to testicular hormones in these mammals. In addition, we suggest that research on urogenital development in these non-traditional species directs attention to processes that may well be operating during the sexual differentiation of morphology and behavior in more common laboratory mammals, albeit in less dramatic fashion.     

Selection underlies phenotypic divergence in the insular Azores woodpigeon

The study of phenotypic evolution in island birds following colonization is a classic topic in island biogeography. However, few studies explicitly test for the role of selection in shaping trait evolution in these taxa. Here, we studied the Azores woodpigeon (Columba palumbus azorica) to investigate differences between island and mainland populations, between females and males, and interactions between geographical origin and sex, by using spectrophotometry to quantify plumage colour and linear measurements to examine external and skeletal morphology. We further tested if selection explains the observed patterns by comparing phenotypic differentiation to genome‐wide neutral differentiation. Our findings are consistent with several predictions of morphological evolution in island birds, namely differences in bill, flight and leg morphology and coloration differences between island and mainland birds. Interestingly, some plumage and morphological traits that differ between females and males respond differently according to geographical origin. Sexual dimorphism in colour saturation is more pronounced in the mainland, but this is driven by selection on female plumage coloration. Differences in flight morphology between females and males are also more pronounced in the mainland, possibly to accommodate contrasting pressures between migration and flight displays. Overall, our results suggest that phenotypic differentiation between mainland and island populations leading to divergent sexual dimorphism patterns can arise from selection acting on both females and males on traits that are likely under the influence of natural and sexual selection.     

Evidence for Sexual Differentiation of Glia in Rat Brain

It is well established that gonadal steroids mediate sexual differentiation of the brain via direct effects on neurons during a restricted critical period. In addition, estrogen can influence glial morphology in the adult brain, andin vitrostudies suggest estrogen induces glial differentiation. However, there is a lack ofin vivoevidence for steroid effects on glia during the critical period. We report here a hormone-mediated sexual differentiation of arcuate glia as early as Postnatal Day 1. Using glial fibrillary acidic protein immunoreactivity (GFAP-ir), we compared the responsiveness of astroglia in the rat arcuate nucleus among five hormonally different groups. The results indicate increased GFAP-ir cell surface area 24 hr after hormonal manipulation in castrate males compared to intact males, intact females (ANOVA;P< 0.01), and females injected with testosterone propionate (50 μg; ANOVA;P< 0.05). However, astroglia in intact males extended their processes significantly greater distances from the cell body compared to all other treatment groups (ANOVA;P< 0.01). The GFAP-ir cells were categorized into four distinct classes ranging from a simple bipolar to a fully stellate morphology. The frequency distribution of classes varied between groups with more stellate cells found in intact males. Finally, these sex differences in arcuate glia persisted into adulthood. We hypothesize that during the critical period, testosterone, or its metabolite estrogen, induce sexual differentiation of glia. We further hypothesize that in females glial cells remain partially undifferentiated and this may be important to glial plasticity seen in adult female arcuate.     

Anthropometric analysis of homosexuals and heterosexuals: implications for early hormone exposure

Early exposure to sex steroids is thought to be important in mediating the differentiation of male-typical sexual orientation. Bone morphology is a marker of childhood sex steroid exposure, because estrogens and androgens control sexual dimorphism in skeletal size. Anthropometric analysis of heterosexuals and homosexuals indicates that those bones, which become sexually dimorphic in childhood, but not those which become sexually dimorphic after puberty, are different in length in homosexuals and heterosexuals. Persons with a sexual preference for males have less long bone growth in the arms, legs and hands, than those with sexual preference for females. The data support the hypothesis that male homosexuals have had less steroid exposure during development than male heterosexuals and that female homosexuals have had greater steroid exposure during development than their heterosexual counterparts.     

用模式生物青鳉概观硬骨鱼性别决定及性分化研究进展

鱼类性别决定和性分化呈多元性,既有雌雄同体也有雌雄异体。性腺的雌雄性分化过程受遗传和环境因素(如温度、光照、激素和pH值等)影响,具有可逆可塑性。随着生物技术和基因组学的迅速发展,近年来脊椎动物性别决定和性分化的研究有了重大进展和显著突破。本文通过聚焦青鳉及其他硬骨鱼纲保守存在的dmrt1、gsdf、amh等雄性因子,探讨硬骨鱼普遍存在的雌雄性别可塑可逆信号通路,并介绍了新的基因组编辑和性控育种技术,为单性选育等水产养殖技术的研发提供参考。     

Gene flow across species boundaries in sympatric, sexually deceptive Ophrys (Orchidaceae) species

Abstract Orchids of the genus Ophrys (Orchidaceae) are pollinated by male bees and wasps through sexual deception. The Ophrys sphegodes group encompasses several closely related species that differ slightly in floral morphology and are pollinated by different solitary bee species. Populations representing different species of the O. sphegodes group often flower simultaneously in sympatry. To test whether gene flow across the species boundaries occurs in these sympatric populations, or whether they are reproductively isolated, we examined the distribution of genetic variation within and among populations and species of this group. We collected at each of five different localities in southern France and Italy two sympatric, co-flowering Ophrys populations, representing six Ophrys species in total. The six microsatellite loci surveyed were highly variable. Genetic differentiation among geographically distant populations of the same species was lower than differentiation among sympatric populations of different species. However, the strength of genetic differentiation among species was among the lowest reported for orchids. Genotype assignment tests and marker-based estimates of gene flow revealed that gene flow across species boundaries occurred and may account for the low observed differentiation among species. These results suggest that sexual deceit pollination in Ophrys may be less specific than thought, or that rare mistakes occur.     

Sexual differentiation in the spiny softshell turtle (Apalone spinifera), a species with genetic sex determination

It is hypothesized on the basis of sex determination theory that species exhibiting genetic sex determination (GSD) may undergo sexual differentiation earlier in development than species with environmental sex determination (ESD). Most turtle species exhibit a form of ESD known as temperature-dependent sex determination (TSD), and in such species the chronology of sex differentiation is well studied. Apalone spinifera is a species of softshell turtle (Trionychidae) that exhibits GSD. We studied sexual differentiation in this species in order to facilitate comparison to TSD species. Eggs were incubated at two different temperatures and embryos were harvested at various stages of mid to late development. Gonad length was measured with image analysis software, then prepared histologically. Indifferent gonads have differentiated in stage 19 embryos. Histological details of gonadogenesis follow the same pattern as described for other reptiles. Regression of the male paramesonephric duct closely follows testicular differentiation. Gonad lengths are longer at the warmer incubation temperature, and ovaries are generally longer than testes at each stage and for each temperature. Although sexual differentiation takes place at about the same stage as in other turtles with TSD (18-20), in A. spinifera this differentiation is irreversible at this stage, while in some of the TSD species sex is reversible until about stage 22. This immutable, definitive sexual differentiation may support the hypothesis of an accelerated chronology of sex differentiation for this species. We also note that sexual dichromatism at hatching is known in this species and may provide additional evidence of early differentiation. J. Exp. Zool. 290:190-200, 2001.     

Distribution of Sexual and Agamospermous Populations of Eupatorium (Compositae) in Asia

Abstract Examinations of chromosome number voucher specimens show that sexual diploid and agamospermous polyploid plants of Eupatorium in Asia can be distinguished by morphology, fertility, and stainability of pollen grains. Using these criteria, reproductive systems (sexual vs. agamospermous) were estimated for 558 herbarium specimens of Eupatorium in East Asia. Of 22 taxa examined, six included both sexual and agamospermous specimens; those of one taxon were all agamospermous; and those of 15 taxa were all sexual. This result shows that 21 taxa are not agamospermous microspecies but are differentiated at the diploid level. Sexual populations of most taxa had restricted geographical distributions. Eupatorium chinense var. chinense and var. oppositifolium consisted of both sexual and agamospermous populations. Sexual populations of these two varieties were allopatric and distinct in external morphology, suggesting differentiation at the species level. The specimens of E. heterophyllum s. str. were all agamospermous, while those of E. mairel , often treated as synonymous with E. heterophyllum , were sexual.
In comparing East Asian and North American Eupatorium for the distributional patterns of sexual and agamospermous populations, three differences can be pointed out: (1) Agamospermous plants of autopolyploid origin have evolved in eight species and are widespread in North America; while most agamospermous plants in East Asia may be of allopolyploid origin, have relatively restricted ranges, and are less frequent than the diploid plants with the exception of in E. chinense var. oppositifolium , (2) The number of sexual species with wide range is greater in North America than in East Asia. (3) The number of sexual species with restricted ranges is greater in East Asia than in North America. The results obtained suggest that speciation among the plants of Eupatorium has occurred more recently in East Asia than in North America.     

Variation in morphology and functional performance across distinct evolutionary lineages of the Moorish gecko (Tarentola mauritanica) from the Iberian Peninsula

Deciphering the mechanisms that underlie morphological and functional diversity is essential for understanding how organisms adapt to their environment. Interestingly, phenotypic divergence does not necessarily correspond to the geographic and genetic separation between populations. Here, we explored the morphological and functional divergence among populations of two genetically differentiated clades of the Moorish gecko, Tarentola mauritanica. We used linear and geometric morphometrics to quantify morphological variation and investigated how it translates into biting and CLIMBING PERFORMANCE, to better understand the mechanisms potentially underlying population and lineage divergence. We found marked morphological differences between clades, both in body size and head shape. However, much of this differentiation is more strongly related to local variation between populations of the same clade, suggesting that recent ecological events may be more influential than deep evolutionary history in shaping diversity patterns in this group. Despite a lack of association between morphology and functional diversification in the locomotor system of the Moorish gecko, straightforward links are observed between head morphology and biting performance, providing more hints on the possible underlying causes. Indeed, variation in bite force is mostly determined by size variation and sexual dimorphism, and differences between the two clades concern how sexual variation is expressed, reinforcing the idea that both social and ecological factors contribute in shaping differentiation. Interestingly, the individuals from the islets off the coast of Murcia exhibit particular morphological and functional traits, which suggests that the ecological conditions related to insularity may drive the phenotypic differentiation of this population.     

High temperatures reveal cryptic genetic variation in a polymorphic female sperm storage organ

Variation in female reproductive morphology may play a decisive role in reproductive isolation by affecting the relative fertilization success of alternative male phenotypes. Yet, knowledge of how environmental variation may influence the development of the female reproductive tract and thus alter the arena of postcopulatory sexual selection is limited. Yellow dung fly females possess either three or four sperm storage compartments, a polymorphism with documented influence on sperm precedence. We performed a quantitative genetics study including 12 populations reared at three developmental temperatures complemented by extensive field data to show that warm developmental temperatures increase the frequency of females with four compartments, revealing striking hidden genetic variation for the polymorphism. Systematic genetic differentiation in growth rate and spermathecal number along latitude, and phenotypic covariance between the traits across temperature treatments suggest that the genetic architecture underlying the polymorphism is shaped by selection on metabolic rate. Our findings illustrate how temperature can modulate the preconditions for sexual selection by differentially exposing novel variation in reproductive morphology. This implies that environmental change may substantially alter the dynamics of sexual selection. We further discuss how temperature-dependent developmental plasticity may have contributed to observed rapid evolutionary transitions in spermathecal morphology.     

Lack of mitochondrial genetic structure in hamlets (Hypoplectrus spp.): recent speciation or ongoing hybridization?

Species in the genus Hypoplectrus (hamlet fish) have been recognized primarily on the basis of colour morphology, which varies substantially. Limited differentiation in other morphological characters, however, has led to ongoing debate about their taxonomy. Our analysis of mtDNA sequences demonstrates neither reciprocal monophyly nor significant genetic differentiation among hamlet colour morphs. These data are potentially consistent with a model of recent speciation due to sexual selection on colour morphology. The presence within hamlets of two divergent mtDNA clades, however, suggests a longer history during which hybridization and gene flow have prevented the differentiation of hamlet colour morphs, at least in neutral genetic markers.     

Studies of gonadal sex differentiation

Gonadal differentiation has a determinative influence on sex development in human embryos. Disorders of sexual development (DSD) have been associated with persistent embryonal differentiation stages. Between 1998 and 2015, 139 female patients with various (DSD) underwent operations at the Scientific Center of Obstetrics, Gynaecology and Perynatology in Moscow, Russia. Clinical investigations included karyotyping, ultrasound imaging, hormonal measurement and investigations of gonadal morphology. The male characteristics in the embryo are imposed by testicular hormones. When these are absent or inactive, the fetus may be arrested at between developmental stages, or stay on indifferent stage and become phenotypically female. A systematic analysis of gonadal morphology in DSD patients and a literature review revealed some controversies and led us to formulate a new hypothesis about sex differentiation. Proliferation of the mesonephric system (tubules and corpuscles) in the gonads stimulates the masculinization of gonads to testis. Sustentacular Sertoli cells of the testes are derived from mesonephric excretory tubules, while interstitial Leydig cells are derived from the original mesenchyme of the mesonephros. According of the new hypothesis, the original mesonephric cells (tubules and corpuscles) potentially persist in the ovarian parenchyma. In female gonads, some mesonephric excretory tubules regress and lose the tubular structure, but form ovarian theca interna and externa, becoming analogous to the sustentacular Sertoli cells in the testis. The ovarian interstitial Leydig cells are derived from intertubal mesenchyme of the mesonephros, similar to what occurs in male gonads (testis). Surprisingly, the leading determinative factor in sexual differentiation of the gonads is the mesonephros, represented by the embryonic urinary system.     

Exaggerated male forelegs are not more differentiated than wing morphology in two widespread sister species of black scavenger flies

Sexual selection represents a potent force that can drive rapid population differentiation in traits related to reproductive success. Hence, sexual traits are expected to show greater population divergence than non-sexual traits. We test this prediction by exploring patterns of morphological differentiation of the exaggerated fore femur (a male-specific sexual trait) and the wing (a non-sexual trait) among allopatric and sympatric populations of the widespread sister dung fly species Sepsis neocynipsea and Sepsis cynipsea (Diptera: Sepsidae). While both species occur in Eurasia, S. neocynipsea also abounds in North America, albeit previous studies suggest strong differentiation in morphology, behavior, and mating systems. To evaluate the degree of differentiation expected under neutrality between S. cynipsea, European S. neocynipsea, and North American S. neocynipsea, we genotyped 30 populations at nine microsatellite markers, revealing almost equal differentiation between and minor differentiation among geographic populations within the three lineages. Landmark-based analysis of 18 populations reared at constant 18 and 24°C in a laboratory common garden revealed moderate temperature-dependent phenotypic plasticity and significant heritable differentiation in size and shape of male forelegs and wings among iso-female lines of the three lineages. Following the biological species concept, there was weaker differentiation between cross-continental populations of S. neocynipsea relative to S. cynipsea, and more fore femur differentiation between the two species in sympatry versus allopatry (presumably due to character displacement). Contrary to expectation, wing morphology showed as much shape differentiation between evolutionary independent lineages as fore femora, providing no evidence for faster diversification of traits primarily engaged in mating.     

栝楼不同性别花芽分化形态解剖特征观察

采用体视显微镜、石蜡切片和树脂切片技术对栝楼(Trichosanthes kirilowii Maxim.)不同性别花芽分化发育时期的外部形态和内部解剖结构进行了观察。结果显示,栝楼花为雌雄异株,仅有雌花、雄花两种性别分化,且雄花的发育速度明显快于雌花的发育速度。栝楼雌雄花芽长0.2 mm左右已完成性别分化;栝楼雄花为单性花,分化过程可分为6个时期,整个发育过程仅见雄蕊原基的分化及生长。栝楼雌花为"两性花",分化过程可分为7个时期,存在雌蕊和雄蕊共同发育阶段,后期雄蕊发育败退。本研究明确了不同性别栝楼花芽发育发生的各个阶段、形态变化特点、外部形态变化特征以及雌雄花芽的分化差异,建立了雌雄花芽内部结构分化与外部形态之间相关性,为栝楼早期幼苗鉴定及性别分化研究提供了一定的参考。     

The Doublesex Locus of Drosophila Melanogaster and Its Flanking Regions: A Cytogenetic Analysis

The region of the third chromosome (84D-F) of Drosophila melanogaster that contains the doublesex (dsx) locus has been cytogenetically analyzed. Twenty nine newly induced, and 42 preexisting rearrangements broken in dsx and the regions flanking dsx have been cytologically and genetically characterized. These studies established that the dsx locus is in salivary chromosome band 84E1-2. In addition, these observations provide strong evidence that the dsx locus functions only to regulate sexual differentiation and does not encode a vital function. To obtain new alleles at the dsx locus and to begin to analyze the genes flanking dsx, 59 lethal and visible mutations in a region encompassing dsx were induced. These mutations together with preexisting mutations in the region were deficiency mapped and placed into complementation groups. Among the mutations we isolated, four new mutations affecting sexual differentiation were identified. All proved to be alleles of dsx, suggesting that dsx is the only gene in this region involved in regulating sexual differentiation. All but one of the new dsx alleles have equivalent effects in males and females. The exception, dsxEFH55, strongly affects female sexual differentiation, but only weakly affects male sexual differentiation. The interactions of dsxEFH55 with mutations in other genes affecting sexual differentiation are described. These results are discussed in terms of the recent molecular findings that the dsx locus encodes sex-specific proteins that share in common their amino termini but have different carboxyl termini. The 72 mutations in this region that do not affect sexual differentiation identify 25 complementation groups. A translocation, T(2;3)Es that is associated with a lethal allele in one of these complementation groups is also broken at the engrailed (en) locus on the second chromosome and has a dominant phenotype that may be due to the expression of en in the anterior portion of the abdominal tergites where en is not normally expressed. The essential genes found in the 84D-F region are not evenly distributed throughout this region; most strikingly the 84D1-11 region appears to be devoid of essential genes. It is suggested that the lack of essential genes in this region is due to the region (1) containing genes with nonessential functions and (2) being duplicated, possibly both internally and elsewhere in the genome.     

Application of organization-activation theory to alternative male reproductive strategies: a review

Many species have extreme within-sex morphological and behavioral polymorphisms, most commonly different male phenotypes that practice different reproductive strategies. Although much is known about the role of hormones in sexual differentiation, little is known about what role hormones might play in within-sex differentiation. The relative plasticity hypothesis is derived from the classical organization-activation model of hormone action. It distinguishes between two types of polymorphic systems: a fixed system in which individual males assume one phenotype for their adult lives and a plastic system in which individual males can change phenotypes at least once. By analogy to sexual differentiation, the relative plasticity hypothesis generally predicts that organizational influences of hormones will be more important in fixed systems and activational influences of hormones will be more important in plastic systems. A review of our knowledge of the role of hormones in differentiation of within-sex polymorphisms indicates that the relative plasticity hypothesis accounts for otherwise diverse and contradictory results. This further supports the hypothesis that the organizational-activational model of hormone action derived from sexual differentiation generalizes to within-sex polymorphisms. However, studies of the effects of hormone manipulations on within-sex differentiation are rare but are desperately needed to further our understanding of this problem. Further studies of discontinuous behavioral variation characteristic of polymorphic species may further our understanding of the physiological basis of within-sex behavior variation in all species.     

Gonadal Hormones and Brain Development: Cellular Aspects of Sexual Differentiation

Sexual differentiation of the neural control of reproductivefunction with respect to both gonadotropin secretion and sexualbehavior is thought to result from exposure of the brain totesticular androgens during a very restricted or critical periodof CNS differentiation and development, when the tissue is competentto respond to the hormone, and after which it is refractoryor responds in a reversible manner. This paper reviews the cellularaspects of sexual differentiation with particular emphasis onthe morphological expression of the gonadal hormonal effectsin the adult brain. It presents experimental evidence for themorphogenetic basis for the observed steroid effects by showinghow the addition of steroid to undifferentiated hypothalamiccultures produces a selective neuritic response that is steroid-dependent.These results suggest that since afferent axonal input and temporalfactors are critical for dendritic and synaptic differentiation,steroid-induced variations in neuritic development could resultin gender-specific responses seen in sexual differentiationof reproductive function.     

The post-natal differentiation of sexual behaviour in the Japanese quail (Coturnix coturnix japonica)

An experiment was carried out to study the contribution of endogenous steroids during postnatal life to the sexual differentiation in Japanese quail. Male and female quail were gonadectomized at one week of age and when adult received testosterone silastic capsules. Seventy seven percent of these females showed male-type sexual behaviour and 46 percent even copulated. All males showed sexual behaviour including cloacal contact movements. Males and females grew cloacal glands of nearly identical sizes in response to the testosterone treatment. It is concluded that sexual differentiation in quail is at least in part a postnatal process and more research is needed to elucidate its exact mechanisms and time course.     

Population differentiation in the swordtail characin (Corynopoma riisei ): a role for sensory drive?

Sensory drive, where the efficacy of a sexual signal depends on the environment in which it is employed, is a potential mechanism behind divergent evolution of secondary sexual traits. Male swordtail characins are equipped with a narrow and transparent extension of the gill cover with a flag‐like structure at its tip. This opercular flag mimics a prey item and is employed by males as a ‘lure’ to attract the attention of females during mating attempts. We conducted a study of genetic and morphological differentiation across swordtail characin populations throughout their native range in Trinidad. The morphology of the opercular flag varied across populations and several aspects of this variation match the predicted hallmarks of sensory drive. First, morphological differentiation of the flag across populations was unrelated to genetic similarity at neutral genetic markers. Second, the shape of the flag covaried with those aspects of body shape that should reflect adaptation to different feeding regimes. Third, and most importantly, the shape of the flag covaried across populations with those environmental characteristics that should most closely reflect differences in local prey abundance. Overall, our results are consistent with a scenario where the evolution of this male sexual signal tracks food‐related shifts in female sensory biases across populations, thus providing at least provisional support for a role for sensory drive in population differentiation.