Genetic Males Research Articles

Sexual differentiation traits in functional males with female genital apertures (♂fga) in the woodlice Armadillidium vulgare Latr. (Isopoda, Crustacea)

This study reports the results of examination of the gonadal morphology and ultrastructural features of the androgenic hormone (AH)-producing androgenic gland cells of laboratory stocks of functional male woodlice, Armadillidium vulgare, with female genital apertures (♂fga), with and without experimentally induced infections of the sex-ratio-distorting endobacterial parasite, Wolbachia. Males (♂fga) have been reported in wild populations containing individuals infected with this maternally transmitted sex-ratio-distorting parasite. We report a reduction of testicular segment (utricle) number, androgenic gland cell hypertrophy, and electron-dense ultrastructural cytological features in ♂fga males. The presence of the Wolbachia parasite had no effect on the features we examined. These results suggest that ♂fga males are produced as the result of a delayed expression/action of the male sex-determining AH which causes a “lag-phase” delay in male differentiation in genetic males and is not due to the presence, in genetic females, of a hypothetical, epigenetic “M” gene as suggested by Rigaud and Juchault [Genetics 133 (1993) 247]. Our results favor the interpretation of males as true genetic (ZZ) males in which the delayed AH action appears to involve cellular AH trafficking pathways which may be controlled by an impaired autosomal gene responsible for AH action.

Evidence for a new feminizing Wolbachia strain in the isopod Armadillidium vulgare: evolutionary implications.

Wolbachia are intracellular maternally inherited alpha-Proteobacteria infecting a wide range of arthropods. In the common pill bug Armadillidium vulgare, the known Wolbachia strain is responsible for feminization of genetic males. We have investigated Wolbachia diversity in 20 populations of A. vulgare from west and east Europe, north Africa and north America. A new Wolbachia strain (wVulM) was identified through the variability of the wsp gene, distantly related to that previously known (wVulC) in this host species. No individual with multiple infections was detected. Inoculation experiments indicated that the new wVulM bacterial strain also induces feminization in A. vulgare. However, the wVulC strain showed a higher transmission rate than the wVulM strain and was the most geographically widespread Wolbachia in A. vulgare populations. Mitochondrial 16SrDNA gene sequencing was conducted in Wolbachia-infected individuals, revealing the occurrence of four host lineages. The comparison of bacterial strains and their respective host mitochondrial phylogenies failed to show concordance, indicating horizontal transmission of the Wolbachia strains within populations of A. vulgare.

Mixed-method research for child outcomes in intersex conditions.

Intersex encompasses a range of inborn errors of metabolism and dysmorphisms, having in common anomalous genitalia. The neonatal diagnosis may be unclear but genital atypia is not. Reconstructive surgery has been the standard of care for more than four decades. Although the conditions are ageless, selfdescriptions of long-term effects are rare, as are the anomalies themselves. For the purposes of this discussion, neonatal genital appearance is meant to reflect inadequate androgen effects in genetic males or inappropriate androgen effects in genetic females. This discussion briefly describes what is known about children’s experiences of and with intersex and intersex treatments, and how such experiences are knowable.

Discordant sexual identity in some genetic males with cloacal exstrophy assigned to female sex at birth.

Cloacal exstrophy is a rare, complex defect of the entire pelvis and its contents that occurs during embryogenesis and is associated with severe phallic inadequacy or phallic absence in genetic males. For about 25 years, neonatal assignment to female sex has been advocated for affected males to overcome the issue of phallic inadequacy, but data on outcome remain sparse. We assessed all 16 genetic males in our cloacal-exstrophy clinic at the ages of 5 to 16 years. Fourteen underwent neonatal assignment to female sex socially, legally, and surgically; the parents of the remaining two refused to do so. Detailed questionnaires extensively evaluated the development of sexual role and identity, as defined by the subjects' persistent declarations of their sex. Eight of the 14 subjects assigned to female sex declared themselves male during the course of this study, whereas the 2 raised as males remained male. Subjects could be grouped according to their stated sexual identity. Five subjects were living as females; three were living with unclear sexual identity, although two of the three had declared themselves male; and eight were living as males, six of whom had reassigned themselves to male sex. All 16 subjects had moderate-to-marked interests and attitudes that were considered typical of males. Follow-up ranged from 34 to 98 months. Routine neonatal assignment of genetic males to female sex because of severe phallic inadequacy can result in unpredictable sexual identification. Clinical interventions in such children should be reexamined in the light of these findings.

Male pseudohermaphroditism: long-term quality of life outcome in five 46,XY individuals reared female.

We assessed the adult quality of life of five medical chart-selected genetic males (ages 29-34 years) assigned and reared as females due to ambiguity of the external genitalia. All five were treated following the traditional method proposed by John Money and colleagues in 1955, commonly referred to as the "optimal gender policy". The adult follow-up assessment included physical and endocrinological evaluation, completion of self-report questionnaires, and a semi-structured interview assessing gender identity, sexual experience and orientation. Quality of life domains assessed by questionnaire included health-related issues, satisfaction with health-care management, emotional distress, and relationship satisfaction. Vaginoplasty in four out of five patients was initially unsuccessful. Four patients had periodic lapses in adherence to hormone replacement therapy. Gender role behavior across development was masculine relative to norms for women. All participants reported a female gender identity without a history of gender dysphoria. The majority of participants (four of five) reported being sexually active and in long-term relationships (three heterosexual, one homosexual). Current emotional adaptation and health-related quality of life are within the normal range for four participants. Sex assignment of 46,XY individuals with ambiguous genitalia as females is compatible with a positive quality of life.

Sexual mosaics induced by tetracycline treatment in the Wolbachia-infected adzuki bean borer, Ostrinia scapulalis.

Wolbachia-infected Ostrinia scapulalis (Lepidoptera: Crambidae) females generate all-female or nearly all-female broods. Curing the infection by tetracycline treatment during larval stages results in the generation of all-male broods in the next generation. Here we show that sexually mosaic offspring are produced by Wolbachia-infected females treated with tetracycline at the adult stage. The sexual mosaics had wings that were composed of distinctive female and male sectors. Besides wings, the sexually dimorphic mid tibiae displayed an intermediate morphology in some of the mosaics. Many of the mosaic individuals had an abnormal structure of the external genitalia as well, a combination of the male uncus and the female ovipositor. We assume that Wolbachia has a feminizing effect on O. scapulalis genetic males and, hence, incomplete curing of the Wolbachia infection results in the generation of sexually mosaic individuals.

Inborn errors of adrenal steroidogenesis

Congenital adrenal hyperplasia (CAH) refers to a family of inherited disorders of adrenal steroidogenesis in which each disorder is characterized by a specific enzyme deficiency that impairs cortisol production by the adrenal cortex. The enzymes most commonly affected are 21-hydroxylase (21-OH), 11β-hydroxylase, 3β-hydroxysteroid dehydrogenase, and less often, 17α-hydroxylase/17,20-lyase and cholesterol desmolase. Many of the corresponding genes for the described enzymes have been isolated and characterized, and specific mutations causing CAH have been identified. In classical CAH (simple virilizing and salt wasting forms), androgen excess causes external genital ambiguity in newborn females and progressive postnatal virilization in both sexes. In nonclassical CAH, 21-OHD is partial and occurs with milder symptoms. A deficiency of 11β-Hydroxylase deficiency results in ambiguous genitalia in the newborn genetic female and androgen excess and hypertension in both males and females. In 3β-hydroxysteroid deficiency adrenal and gonadal androgen production is deficient resulting in incomplete genital development in genetic males and limited androgen affect in females. Two less frequent causes of CAH 17α-Hydroxylase/17,20-lyase and cholesterol desmolase result in external female genitalia in both sexes. Hormonal diagnosis is described for each disorder.

XX MALE: A RARE CAUSE OF SHORT STATURE, INFERTILITY, GYNAECOMASTIA AND CARCINOMA OF THE BREAST

SUMMARYWe present a case of an XX male with carcinoma of the breast and primary infertility. He was admitted to hospital with recurrent chest pains, but a history of surgery for breast carcinoma, gynaecomastia and the finding of bilaterally atrophied testes, coupled with the fact that he had never fathered children, necessitated further investigations. Chromosomal analysis showed a 46, XX male genotype with a normal X chromosome and an abnormal X chromosome formed by translocation between the short arm of one X chromosome and the Y chromosome. By using fluorescence in situ hybridisation, the patient proved to be SRY positive, the sex‐determining region of the Y chromosome. In this rare genetic abnormality males retain normal phenotype but they are generally of short stature, have gynaecomastia, and may have genital anomalies. They are infertile and at increased risk of developing carcinoma of the breast. This seems to be the first documented case of carcinoma of the breast in an SRY positive XX male. This particular case illustrates the need for all cases of male breast cancer to undergo full endocrinological assessment, especially in the presence of genital anomaly or infertility. (Int J Clin Pract 2003; 57(9): 844–845)

Microorganisms and parthenogenesis

Endosymbionts are being discovered in unprecedented numbers of invertebrate species. The bacterium Wolbachia pipientis alone is carried by some 20% of all insect species, in which it is responsible for such reproductive manipulations as cytoplasmic incompatibility, feminization of genetic males, male-killing and parthenogenesis. The discovery of Wolbachia-induced thelytoky in hymenopteran parasitoids has initiated the successful search in other groups of haplodiploid species. Haplodiploidy has arisen at least 17 times during animal evolution and intracellular bacteria that effect sexual development have been found in many of these haplodiploid groups, rekindling the question regarding the origin of haplodiploidy. The detection of Wolbachia in parthenogenetic thrips (Thysanoptera). booklice (Psocoptera), springtails (Collembola), mites (Acari) and Xiphinematobacter in dragger nematodes (Dorylaimida) raises the possibility of bacterial-induced parthenogenesis in diplodiploid species as well. Recent studies indicate that Wolbachia might be just one of a rapidly increasing list of intracellular bacteria capable of inducing parthenogenesis in their host. The search for novel endosymbionts will rely heavily on previous taxonomic work, but the possibilities for discovery remain boundless.

Dietary ethinyl estradiol exposure during development causes increased voluntary sodium intake and mild maternal and offspring toxicity in rats

Exogenous estrogen exposure during development often results in behavioral masculinization and/or defeminization of genetic females. Genetic males may be defeminized, hypermasculinized or even demasculinized after similar treatment. Here, pregnant Sprague–Dawley rats consumed phytoestrogen-free diets containing 0, 1, 5 or 200 ppb EE 2 beginning on gestational day (GD) 7. Offspring were weaned to the same maternal diet and maintained gonadally intact. There were mild effects on body weight and food consumption in dams of the 200 ppb group and their offspring weighed less at birth than those of the control group; however, gross assessments of nursing behavior were normal in all dietary groups. Postweaning, offspring of the 200 ppb group weighed less and consumed less food than controls. There were no EE 2-related effects on open-field activity (tested at postnatal days (PND) 22–24, 43–45 and 64–66), play behavior (tested at PND 35), running wheel activity (PND 63–77) or intake of a 0.3% saccharin-flavored solution (PND 69–71). Intake of a 3.0% sodium chloride-flavored solution on PND 73–75 was increased in both male and female offspring of the 200 ppb group relative to same-sex controls, an effect that is reportedly estrogen mediated. Sodium chloride-flavored solution intake is a sexually dimorphic behavior for which female rats consume more than males. Here, while EE 2 exposure had few effects on the conventional tests of sexually dimorphic behaviors, exposure to 200 ppb in the diet appeared to feminize genetic males and hyperfeminize genetic females with regard to sodium intake.

Neonatal sex steroids affect responses to Seoul virus infection in male but not female Norway rats

Previous studies illustrate that after inoculation with Seoul virus (i.e., the naturally occurring hantavirus found in Norway rats), adult male rats produce higher antibody responses, exhibit higher Th1 responses (i.e., IgG2a, IL-2, and IFNγ), and shed virus longer than females, but these difference are not altered by manipulation of sex steroids in adulthood. To determine whether sex steroid hormones organize adult responses to hantavirus infection, at 2–4 days of age, male rats were gonadectomized and female rats were injected with testosterone. As adults, animals were inoculated with 10 4pfu of Seoul virus. Neonatally gonadectomized males (NGM), control females (CF), and neonatal testosterone-treated females (NTF) had lower anti-Seoul virus IgG and IgG2a responses than control males (CM) 20, 30, and 40 days post-inoculation. Neonatal testosterone treatment had no effect on female antibody responses to infection. NGM, CF, and NTF shed virus in saliva and feces for a shorter duration than CM. There was no effect of neonatal hormone manipulation either on the percentage of animals with detectable virus or on the number of virus copies within each target organ. Genetic males, regardless of hormone manipulation, had higher virus replication in lung tissue than did genetic females. Neonatal sex steroids influence the sexual dimorphism in host immune function but do not modify virus replication in target tissues.

Y-chromosomal DNA markers for discrimination of chemical substance and effluent effects on sexual differentiation in salmon.

Chinook salmon alevins were exposed during their labile period for sex differentiation to different concentrations of bleached kraft mill effluent (BKME), primary sewage effluent, secondary sewage effluent (SE), 17ss-estradiol, testosterone, and nonylphenol. After exposure for 29 days post hatching (DPH), fish were allowed to grow until 103 and 179 DPH, at which time their genetic sex was determined using Y-chromosomal DNA markers and their gonadal sex was determined by histology. Independent of treatment, all fish identified as genetic females (XX) in these experiments possessed normal female gonads. Only the highest dose of some treatments affected the development of gonads in genetic XY males. At 103 DPH, some genetic males treated with 1 micro g estradiol/L, BKME 100%,and SE 30% developed as physiological females, presenting ovaries identical to genetic females in the control group. The physiological female condition in XY fish was also observed in these treatments groups at 179 DPH, which suggests that the effect is permanent, whereas in other groups the effect changed between sampling periods. Identification of the genetic sex of individual animals using sex-linked DNA markers provides a useful tool for investigating environmental factors influencing sex determination and differentiation.

Incidence of Phenotypic Female Chinook Salmon Positive for the Male Y-Chromosome-Specific MarkerOtY1in the Central Valley, California

Abstract Fall-run chinook salmon Oncorhynchus tshawytscha collected during 1999 carcass surveys of seven locations within the Sacramento River basin and six locations within the San Joaquin River basin, California, were screened using the Y-chromosome-specific marker OtY1 to evaluate inconsistencies between sexual genotype and phenotype. Of 287 phenotypic females screened, 46 (16% overall) tested positive for the Y-chromosome marker. Stream populations had higher frequencies of sex-reversed males (American River, 20%; Battle Creek, 35%; Feather River, 20%; Merced River, 24%; Mokelumne River, 38%; and Yuba River, 25%) than hatchery populations (Feather River Hatchery, 0%; Nimbus Hatchery, 12%; Merced River Hatchery, 14%; and Mokelumne River Hatchery, 4%), and several other streams had intermediate frequencies. All 150 male fall-run chinook salmon from the sampling locations tested positive for the Y-chromosome marker. These results present evidence that some genetic males have been sex reversed and have th...

Feminizing Wolbachia in an insect, Ostrinia furnacalis (Lepidoptera: Crambidae).

Wolbachia, which forms a group of maternally inherited bacteria in arthropods, often cause reproduction alterations in their hosts, such as cytoplasmic incompatibility, parthenogenesis, male-killing, hybrid breakdown and feminization. To date, Wolbachia-induced feminization has been reported only in isopods. Here we report that a Wolbachia strain feminizes an insect host, Ostrinia furnacalis. Among 79 wild females of O. furnacalis examined, Wolbachia infection was detected in 13 females. Twelve of the 13 infected females produced all-female progenies, and this trait was maternally inherited. Tetracycline treatment of thelygenic matrilines resulted in the production of all-male progenies. The present findings indicate that the Wolbachia infection induces feminization of genetic males in O. furnacalis. Differences in the Wolbachia-induced feminization in O. furnacalis and that in isopods are discussed along with the differences in sex determination mechanisms between insects and isopods. Phylogenetic analysis of the wsp gene sequence of Wolbachia suggests independent evolutionary origins for the Wolbachia-induced feminizations in O. furnacalis and in isopods. Our findings over 5 years suggest that the infection has been maintained at a low prevalence in the O. furnacalis population.

Feminization of genetic males by a symbiotic bacterium in a butterfly, Eurema hecabe (Lepidoptera: Pieridae).

Wolbachia are symbiotic bacteria found in many arthropods and filarian nematodes. They often manipulate the reproduction of host arthropods. In the present study, female-biased sex-ratio distortion in the butterfly Eurema hecabe was investigated. Breeding experiments showed that this distorted sex ratio is maternally inherited. When treated with tetracycline, adult females of the thelygenic line produced male progeny only. After PCR using Wolbachia-specific primers for the ftsZ gene a positive result was seen in the thelygenic females, but not in male progeny from tetracycline-treated females, or individuals from a Tokyo population with normal sex ratio and reproduction. Cytological observations showed that thelygenic females lack the sex chromatin body (W chromosome). The results strongly suggest that the sex-ratio distortion in E. hecabe is due to feminization of genetic males by Wolbachia.

Haploid all the way: a new style of asexuality revealed in animals.

Weeks et al(1) recently reported that they had found a species of mites where the parthenogenetic females are haploid. They show that this is caused by intracellular bacteria that turn genetic haploid males into haploid females. I discuss these findings and attempt to place these observations in evolutionary context.

Congenital adrenal hyperplasia due to 3beta-hydroxysteroid dehydrogenase/Delta(5)-Delta(4) isomerase deficiency.

The 3beta-hydroxysteroid dehydrogenase/Delta(5)-Delta(4)isomerase (3beta-HSD) isoenzymes are responsible for the oxidation and isomerization of Delta(5)-3beta-hydroxysteroid precursors into Delta(4)-ketosteroids, thus catalyzing an essential step in the formation of all classes of active steroid hormones. The 3beta-HSD gene family should have evolved to facilitate differential patterns of tissue- and cell-specific expression and regulation involving multiple signal transduction pathways, which are activated by several growth factors, steroids, and cytokines. In humans, there are two 3beta-HSD isoenzymes, which were chronologically designated type I and II encoded by HSD3B1 and HSD3B2 gene, respectively. HSD3B1 gene encodes the almost exclusive 3beta-HSD isoenzyme expressed in the placenta and peripheral tissues, whereas HSD3B2 gene encodes the predominant 3beta-HSD isoenzyme expressed in the adrenal gland, ovary, and testis and its deficiency is responsible for a rare form of congenital adrenal hyperplasia causing various degrees of salt-wasting in both sexes and incomplete masculinization of the external genitalia in genetic males. Although an elevated ratio of Delta(5)-Delta(4)-steroids was considered to be the best biological parameter for the diagnosis of this autosomal recessive disorder, the most accurate criteria now appears to be the plasma levels of 17-OH-pregnenolone greater than 100 nmol/L following ACTH stimulation. To date a total of 34 mutations (including 5 frameshift, 4 nonsense, 1 in-frame deletion, 1 splicing, and 23 missense mutations) have been identified in the HSD3B2 gene in 56 individuals from 44 families suffering from classical 3beta-HSD deficiency. In almost all the cases, the functional characterization of HSD3B2 mutations has provided a molecular explanation for the heterogeneous clinical presentation of this disorder. Indeed these experiments confirm that no functional 3betaHSD type II isoenzyme is expressed in the adrenals and gonads of the patients suffering from a severe salt-wasting form, whereas the non-salt-losing form results from specific missense mutation(s) in the HSD3B2 gene, which causes an incomplete loss of enzymatic activity thus leaving sufficient enzymatic activity to prevent salt wasting. Moreover, various mutations appear to have a drastic effect upon stability of the protein, therefore providing molecular evidence of a new mechanism involved in classical 3beta-HSD deficiency. Thus, the elucidation of the molecular basis of 3beta-HSD deficiency has highlighted the fact that mutations in the HSD3B2 gene can result in a wide spectrum of molecular repercussions, which are associated with the different phenotypic manifestations of classical 3beta-HSD deficiency and also provide valuable information concerning the structure-function relationships of the 3beta-HSD superfamily.

CHOLESTEROL AND DEVELOPMENT: THE RSH ("SMITH-LEMLI-OPITZ") SYNDROME AND RELATED CONDITIONS

The half-century of lipophobia in the United States may be abating with some return of sanity on the discussion of health and dietary fat [Taubes, 2001]. The youngest victims of this collective, decades long madness are those infants deprived for one reason or another of breast milk. They are unable to speak for themselves at a time of greatest need for cholesterol during growth, the most critical period of myelination of central and peripheral nervous system, formation of bone and bile, and of every steroid hormone. Some of the commercial formulas they are fed contain only 1 or 2 mg of cholesterol per 100 g edible portion contrasted with almost 14 mg in breast milk. One can only hope that the confidence in their endogenous ability to synthesize sufficient amounts of cholesterol is not misplaced. Pediatric pathology has learned that when this endogenous ability fails during embryogenesis on the basis of mutations in the postsqualene biosynthesis of cholesterol, a startling variety of developmental pathology may present itself ranging from lethal forms of "idiopathic" hydrops, microcephaly with cerebral dysgenesis and dysmyelinization, agenesis of corpus callosum, cerebellar vermis dysgenesis, cataracts, cleft palate, many different forms of congenital heart defect, pyloric stenosis and/or Hirschsprung dysganglionosis, adrenal (cortical) insufficiency, cholestatic liver disease, limb malformations, and genital ambiguity in genetic males. Population genetic considerations suggest a hypothetical birth prevalence of the RSH (so-called Smith-Lemli-Opitz) syndrome, the commonest of these Garrodian errors of cholesterol biosynthesis, of 1/2500; since only about 1/15,000 to 1/20,000 homozygotes are liveborn and biochemically confirmed, over 80% prenatal or perinatal mortality must occur and deserves the most discerning of services from birth attendants, perinatologists, neonatologists, and fetal/pediatric pathologists. An easy, reliable, economical biochemical test for the presence of 7-dehydrocholesterol is available and the commonest mutation, the IVS8-1G &#77 C mutation, is quickly and reliably tested for molecularly. Thus, the successful diagnosis, even after death, will contribute substantially to correct genetic counseling, carrier detection, prenatal diagnosis, and treatment in those known to be affected prenatally and planned to be liveborn. Thus, developmental pathology plays an integral, vital role in preventive medicine.

Disorders of androgen biosynthesis.

Disorders of androgen biosynthesis are a relatively rare cause of sexual ambiguity in 46,XY genetic males, but genetic disorders characterized by excessive androgen synthesis are a common cause of virilization in 46,XX genetic females. Understanding of these disorders is relatively simple if one is familiar with the components of the various steroidogenic pathways. The biosynthesis of androgens requires the steroidogenic acute regulatory protein (StAR) and the steroidogenic enzymes P450scc, P450c17, 3betaHSDII, 17betaHSDIII, and 5alpha-reductase. Deficiencies have been described in all of these, leading to male pseudohermaphroditism. Other steroidogenic enzymes not involved in androgen biosynthesis, such as P450c21, P450c11beta, and P450aro, must also be considered, as mutations in these can result in overproduction of androgens, resulting in female pseudohermaphroditism.

Cloning and characterization of a gene encoding glutathione-regulated potassium-efflux system protein KefKL from the endosymbiont Wolbachia.

The maternally inherited intracellular symbiont Wolbachia is well known for inducing a variety of reproductive and developmental abnormalities in the diverse arthropod hosts it infects. It has been implicated in causing cytoplasmic incompatibility (CI), parthenogenesis, feminization of genetic males and male killing in different hosts. However, the molecular mechanisms by which this fastidious bacterium causes these abnormalities have not yet been determined. In our study, representational difference analysis (RDA) was used to analyze the genomic difference between different Wolbachia strains. A gene encoding glutathione-regulated potassium-efflux system protein KefKL from Wolbachia in Drosophila simulans Riverside (w Ri) was isolated. The homologous genes from Wolbachia in Drosophila melanogaster yw67c23 (wMel) and Wolbachia in Drosophila melanogaster CantonS (wMelCS) were also cloned and sequenced. Sequence analysis showed that these deduced amino acid sequences contained two important motifs: Na+/H+ antiportor and NAD binding domain, which shared conserved sequences among different strains. Considering the crucial function of KefKL for ionic homeostasis, this gene might play an important role in Wolbachia physiology. Further study indicated that there was no homologue detected from Wolbachia in Drosophila simulans DSW/Mau (wMa) and Wolbachia in Drosophila simulans Noumea (wNo). Whether Wolbachia contained KefKL (or the homologous gene) was consistent with the phylogenetic studies using wsp sequences, which showed that wMa and wNo were grouped into one branch, while w Ri, wMel and wMelCS were more closely related.