Sexual differentiation and gonad development in striped mullet (Mugil cephalus L.) from South Carolina estuaries*

This study examined the sexual differentiation and reproductive dynamics of striped mullet (Mugil cephalus L.) in the estuaries of South Carolina. A total of 16,464 specimens were captured during the study and histological examination of sex and maturity was performed on a subsample of 3670 fish. Striped mullet were sexually undifferentiated for the first 12 months, began differentiation at 13 months, and were 90% fully differentiated by 15 to 19 months of age and 225 mm total length (TL). The defining morphological characteristics for differentiating males was the elongation of the protogonial germ tissue in a corradiating pattern towards the center of the lobe, the development of primary and secondary ducts, and the lack of any recognizable ovarian wall structure. The defining female characteristics were the formation of protogonial germ tissue into spherical germ cell nests, separation of a tissue layer from the outer epithelial layer of the lobe-forming ovarian walls, a tissue bud growing from the suspensory tissue that helped form the ovary wall, and the proliferation of oogonia and oocytes. Sexual maturation in male striped mullet first occurred at 1 year and 248 mm TL and 100% maturity occurred at age 2 and 300 mm TL. Female striped mullet first matured at 2 years and 291 mm total length and 100% maturity occurred at 400 mm TL and age 4. Because of the open ocean spawning behavior of striped mullet, all stages of maturity were observed in males and females except for functionally mature females with hydrated oocytes. The spawning season for striped mullet recruiting to South Carolina estuaries lasts from October to April; the majority of spawning activity, however, occurs from November to January. Ovarian atresia was observed to have four distinct phases. This study presents morpholog ical analysis of reproductive ontogeny in relation to size and age in South Carolina striped mullet. Because of the length of the undifferentiated gonad stage in juvenile striped mullet, previous studies have proposed the possibility of protandric hermaphrodism in this species. The results of our study indicate that striped mullet are gonochoristic but capable of exhibiting nonfunctional hermaphroditic characteristics in differentiated mature gonads.

Long-term toxic impact of 2,3,7,8-tetrachlorodibenzo-p-dioxin on the reproduction, sexual differentiation, and development of different life stages of Gobiocypris rarus and Daphnia magna

The sexual ratio of Gobiocypris rarus exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and 17 beta -estradiol from embryo to sexually mature revealed feminization and overdevelopment of connective tissue in male fish gonad in 2-30 pg/L TCDD concentration range. Daphnia magna was not sensitive to the high dose of TCDD (0.1-1000 ng/ml), but the reproduction of D. magna treated with TCDD decreased after the 8th day. 7-Ethoxyresorufin-O-deethylase (EROD) activities in newly fertilized eggs of G. rarus exposed to TCDD dosage groups (1000-100,000 pg/L) were significantly induced and increased with TCDD concentrations at the early life stage, while no difference was found between low TCDD dosage groups (<100 pg/L), but a good relationship between the EROD activity and the TCDD concentration was observed during a long-term developmental stage. There was a pericardial edema formed in a 2-week yolk-sac at the concentration of 1000 pg/L TCDD. But in the exposure group (2 pg/L TCDD for 120 days), the cell nuclei of hepatocytes was far from the center and packed toward the cell membrane; the cristae of most mitochondria in the cell dropped and collapsed; the rough endoplasmic reticulum broke into fragments; and numerous lipid droplets formed in the cell. (C) 2001 Academic Press.

Endocrine Disruptors and Hypothalamic Sexual Differentiation

The so-called endocrine disruptors have been described as compounds which interfere with the estrogen action in their receptors and may exert a crucial role in the development of the reproductive tract and in the brain sexual differentiation. Thus, conducts and/or exposure to these drugs in the perinatal period that apparently do not endanger the neonate may cause side effects. During embrionary development, the gonads, through discharge of a small quantity of reproductive hormones, will guarantee the phenotype of male or female at birth, as well as actuate in specific areas sexual differentiation of the central nervous system. Several experimental models have shown an interference of drugs acting as endocrine disruptors in hypothalamic sexual differentiation. Thus, reproductive function is impaired by exposure to estrogen in the perinatal life of rats and the mechanisms involved in this effect are distinct for males and females. Perinatal exposure to drugs which may be considered endocrine disrupters may induce an incomplete masculinization and defeminization of the central nervous system. Alterations in these processes, if present, generally are perceived only at puberty or adult reproductive life. These later alterations may include anomalies in the process of fertility or in sexual behavior.

Why boys will be boys: two pathways of fetal testicular androgen biosynthesis are needed for male sexual differentiation

Human sexual determination is initiated by a cascade of genes that lead to the development of the fetal gonad. Whereas development of the female external genitalia does not require fetal ovarian hormones, male genital development requires the action of testicular testosterone and its more potent derivative dihydrotestosterone (DHT). The "classic" biosynthetic pathway from cholesterol to testosterone in the testis and the subsequent conversion of testosterone to DHT in genital skin is well established. Recently, an alternative pathway leading to DHT has been described in marsupials, but its potential importance to human development is unclear. AKR1C2 is an enzyme that participates in the alternative but not the classic pathway. Using a candidate gene approach, we identified AKR1C2 mutations with sex-limited recessive inheritance in four 46,XY individuals with disordered sexual development (DSD). Analysis of the inheritance of microsatellite markers excluded other candidate loci. Affected individuals had moderate to severe undervirilization at birth; when recreated by site-directed mutagenesis and expressed in bacteria, the mutant AKR1C2 had diminished but not absent catalytic activities. The 46,XY DSD individuals also carry a mutation causing aberrant splicing in AKR1C4, which encodes an enzyme with similar activity. This suggests a mode of inheritance where the severity of the developmental defect depends on the number of mutations in the two genes. An unrelated 46,XY DSD patient carried AKR1C2 mutations on both alleles, confirming the essential role of AKR1C2 and corroborating the hypothesis that both the classic and alternative pathways of testicular androgen biosynthesis are needed for normal human male sexual differentiation.

Of marsupials and men: "Backdoor" dihydrotestosterone synthesis in male sexual differentiation

Following development of the fetal bipotential gonad into a testis, male genital differentiation requires testicular androgens. Fetal Leydig cells produce testosterone that is converted to dihydrotestosterone in genital skin, resulting in labio-scrotal fusion. An alternative 'backdoor' pathway of dihydrotestosterone synthesis that bypasses testosterone has been described in marsupials, but its relevance to human biology has been uncertain. The classic and backdoor pathways share many enzymes, but a 3α-reductase, AKR1C2, is unique to the backdoor pathway. Human AKR1C2 mutations cause disordered sexual differentiation, lending weight to the idea that both pathways are required for normal human male genital development. These observations indicate that fetal dihydrotestosterone acts both as a hormone and as a paracrine factor, substantially revising the classic paradigm for fetal male sexual development.

The zebrafish, brain-specific, aromatase cyp19a2 is neither expressed nor distributed in a sexually dimorphic manner during sexual differentiation

Differential cyp19 aromatase expression during development leads to sexual dimorphisms in the mammalian brain. Whether this is also true for fish is unknown. The aim of the current study has been to follow the expression of the brain-specific aromatase cyp19a2 in the brains of sexually differentiating zebrafish. To assess the role of cyp19a2 in the zebrafish brain during gonadal differentiation, we used quantitative reverse transcriptase-polymerase chain reaction and immunohistochemistry to detect differences in the transcript or protein levels and/or expression pattern in juvenile fish, histology to monitor the gonadal status, and double immunofluorescence with neuronal or radial glial markers to characterize aromatase-positive cells. Our data show that cyp19a2 expression levels during zebrafish sexual differentiation cannot be assigned to a particular sex; the expression pattern in the brain is similar in both sexes and aromatase-positive cells appear to be mostly of radial glial nature.

In vivo definition of genetic and cellular aspects of mammalian sexual differentiation

The differentiation of the reproductive organs is an essential developmental process required for the proper transmission of the genetic material. Müllerian inhibiting substance (MIS) is produced by testes and is necessary for the regression of the Müllerian ducts: the anlagen of the uterus, fallopian tubes and cervix. In vitro and standard transgenic mouse studies indicate that the nuclear hormone receptor Steroidogenic factor 1 (SF-1) and the transcription factor SOX9 play an essential role in the regulation of Mis. To test this hypothesis, mutations in the endogenous SF-1 and SOX9 binding sites in the mouse Mis promoter were introduced by gene targeting in embryonic stem (ES) cells. In disagreement with cell culture and transgenic mouse studies, male mice homozygous for the mutant SF-1 binding site correctly initiated Mis transcription in the fetal testes, although at significantly reduced levels. Surprisingly, sufficient Mis was produced for complete elimination of the Müllerian duct system. However, when the SF-1 binding site mutation was combined with an Mis -null allele, the further decrease in Mis levels led to a partial retention of uterine tissue, but only at a distance from the testes. In contrast, males homozygous for the mutant SOX9 binding site did not initiate Mis transcription, resulting in pseudohermaphrodites with a uterus and oviducts. These studies suggest an essential role for SOX9 in the initiation of Mis transcription, whereas SF-1 appears to act as a quantitative regulator of Mis transcript levels perhaps for influencing non-Müllerian duct tissues. ^ The Mis type II receptor, a member of the TGF- b superfamily, is also required for the proper regression of the Müllerian ducts. Mis type II receptor-deficient human males and their murine counterparts develop as pseudohermaphrodites. A lacZ reporter cassette was introduced into the mouse Mis type II receptor gene, by homologous recombination in ES cells. Expression studies, based on b -galactosidase activity, show marked expression of the MIS type II receptor in the postnatal Sertoli cells of the testis as well as in the prenatal and postnatal granulosa cells of the ovary. Expression is also seen in the mesenchymal cells surrounding the Müllerian duct and in the longitudinal muscle layer of the uterus. ^

Estrogen receptor-dependent sexual differentiation of dopaminergic neurons in the preoptic region of the mouse

Although it has been known for some time that estrogen exerts a profound influence on brain development a definitive demonstration of the role of the classical estrogen receptor (ERα) in sexual differentiation has remained elusive. In the present study we used a sexually dimorphic population of dopaminergic neurons in the anteroventral periventricular nucleus of the hypothalamus (AVPV) to test the dependence of sexual differentiation on a functional ERα by comparing the number of tyrosine hydroxylase (TH)-immunoreactive neurons in the AVPV of wild-type (WT) mice with that of mice in which the ERα had been disrupted by homologous recombination (ERKOα). Only a few ERα-immunoreactive neurons were detected in the AVPV of ERKOα mice, and the number of TH-immunoreactive neurons was three times that of WT mice, suggesting that disruption of the ERα gene feminized the number of TH-immunoreactive neurons. In contrast, the AVPV contains the same number of TH-immunoreactive neurons in testicular feminized male mice as in WT males, indicating that sexual differentiation of this population of neurons is not dependent on an intact androgen receptor. The number of TH-immunoreactive neurons in the AVPV of female ERKOα mice remained higher than that of WT males, but TH staining appeared to be lower than that of WT females. Thus, the sexual differentiation of dopamine neurons in the AVPV appears to be receptor specific and dependent on the perinatal steroid environment.

Excitatory versus inhibitory GABA as a divergence point in steroid-mediated sexual differentiation of the brain

Whereas adult sex differences in brain morphology and behavior result from developmental exposure to steroid hormones, the mechanism by which steroids differentiate the brain is unknown. Studies to date have described subtle sex differences in levels of proteins and neurotransmitters during brain development, but these have lacked explanatory power for the profound sex differences induced by steroids. We report here a major divergence in the response to injection of the γ-aminobutyric acid type A (GABAA) agonist, muscimol, in newborn male and female rats. In females, muscimol treatment primarily decreased the phosphorylation of cAMP response element binding protein (CREB) within the hypothalamus and the CA1 region of the hippocampus. In contrast, muscimol increased the phosphorylation of CREB in males within these same brain regions. Within the arcuate nucleus, muscimol treatment increased the phosphorylation of CREB in both females and males. Thus, the response to GABA can be excitatory or inhibitory on signal-transduction pathways that alter CREB phosphorylation depending on the sex and the region in developing brain. This divergence in response to GABA allows for a previously unknown form of steroid-mediated neuronal plasticity and may be an initial step in establishing sexually dimorphic signal-transduction pathways in developing brain.

Controle hormonal da diferenciação e inversão sexual na dourata Sparus aurata (L.)

Tese de Doutoramento, Biologia Marinha, Unidade de Ciências e Tecnologias dos Recursos Aquáticos, Universidade do Algarve, 2001

Interação sexual: perspectiva etológica do dimorfismo sexual humano

A interação sexual no sentido de ação conjunta ou recíproca supõe a existência de parceiros dimorficamente caracterizados a partir da estrutura cromossomial X-X ou X-Y. A diferenciação do sexo, entretanto, não depende apenas da condição cromossomial, podendo mesmo ser invertida quando a ação hormonal for alterada. Os dados disponíveis apontam os andrógenos como um dos principais agentes de diferenciação, afetando o organismo "básico" feminino num sentido masculinizante. A medida que se progride na escala animal, contudo, os aspectos estritamente biológicos perdem em importância e as condições ambientais aos poucos se impõem . Nos primatas superiores, caso não se estabeleçam condições adequadas de aprendizagem, o desenvolvimento sexual não se completa e a cópula se torna inviável. No ser humano, além dessa aprendizagem, fatores socio-culturais respondem pelo comportamento considerado masculino ou feminino que supostamente será exibido por cada parceiro da interação. Também as noções de estética são seriamente determinadas pela tradição cultural, com cada povo tendendo a valorizar os próprios traços raciais como ideal de beleza. Do ponto de vista físico, entretanto, pode-se tentar estabelecer algumas constantes, como as formas arredondadas para a mulher e o porte atl6tico ou capacidades gerais , para o homem. Uma vez estabelecida a atração e encontrado local e momento adequado, a interação sexual tende a uma progressiva intensificação e genitalização, que, nas sociedades de tradição judaico-cristã se reflete numa sequência, mais ou menos previsível, envolvendo carícias gerais, beijos, acariciamento de seios e genitais até a união genito-genital o significado e a forma como cada cultura desenvolve os contatos físicos de intercambio erógeno revelam tão grande variedade que já se apontou essa polimorfismo corno aquisição típica do ser humano. Assim, ao contrário do que faz supor o ideal de abstinência, é na capacidade de buscar e estender o prazer além da s1mples atividade orgânico-sexual que o homem, em sexo, se liberta da limitação animal.

Adult partner preference and sexual behavior of male rats exposed prenatally to betamethasone

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)