Beta-tubulin mutants of the unicellular green alga Chlamydomonas reinhardtii.

Two beta-tubulin mutants of the unicellular green alga Chlamydomonas reinhardtii have been isolated on the basis of altered sensitivity to the growth-inhibitory effect of colchicine. The two mutations of colR4 and colR15 have been found to be tightly linked, mapping to a previously unmarked site in linkage group XII. The drug-resistance phenotypes of both mutations segregated in genetic crosses with the presence of distinct, acidic variant beta-tubulin isoforms found assembled into the microtubules of the flagella. Analysis of the in vitro translation products of total poly(A)+ RNA from the mutants provided evidence that the variant proteins are altered primary beta-tubulin gene products. Compared to wild type, strains carrying the mutations expressed an increased resistance to the inhibitory effects of colchicine in clonal growth, flagellar assembly, and germination of meiotic products, suggesting that the beta-tubulin altered in the mutants participates in multiple microtubule functions.     

Two-dimensional analysis of flagellar proteins from wild-type and paralyzed mutants of Chlamydomonas reinhardtii.

Flagellar polypeptides of Chlamydomonas reinhardtii were analyzed in two-dimensions by isoelectric focusing and electrophoresis in the presence of sodium dodecyl sulfate. In addition to flagellar tubulin, over 130 polypeptides were resolved and 100 of these were identified as axonemal components in wild-type organisms. Flagella of two nonconditional paralyzed mutants, pf 14 and pf 1, were also analyzed and, at the same time, electron microscopic studies were carried out. pf 14 flagella, which completely lack radial spokes and associated spokeheads, are missing 12 polypeptides. Six of these polypeptides are also missing from pf 1 flagella in which spokes are clearly present but spoke heads appear to be absent.     

Mammalian sexual differentiation: lessons from the spotted hyena.

Female spotted hyenas (Crocuta crocuta) are the only female mammals that lack an external vaginal opening. Mating and birth take place through a urogenital canal that exits at the tip of a hypertrophied clitoris. This 'masculine' phenotype spurred a search for an alternate source of fetal androgens. Although androstenedione from the maternal ovary is readily metabolized to testosterone by the hyena placenta, formation of the penile clitoris and scrotum appear to be largely androgen independent. However, secretions from the fetal testes underlie sex differences in the genitalia and central nervous system that are essential for male reproduction. Naturally circulating androgens, acting prenatally, reduce reproductive success in adult female spotted hyenas. Effects on aggression and dominance might offset these reproductive 'costs' of female androgenization in utero.     

On the obligatory role of the hypophysis in sexual differentiation hepatic metabolism in rats.

The hepatic metabolism of 4-[4-14C]androstene-3,17-dione and 5alpha-[4-14C]androstane-3alpha,17beta-diol was studies in castrated and hypophysectomized male rats with a transplanted pituitary under the kidney capsule. The effects of testosterone propionate and estradiol benzoate on liver metabolism were also studied in these experimental animals. It was found that the autonomous pituitary secreted a "feminizing factor" that transformed the male type of steroid metabolism characteristic of hypophysectomized rats into a female type of metabolism Hypophysectomized rats were unresponsive towards androgen action on the liver and did not respond with feminized hepatic metabolism after treatment with estradiol benzoate. It is concluded that estrogenic action on liver enzymes is mediated via modulation of the secretion of a central (probably hypothalamic) "feminizing factor inhibiting factor" and the sex hormonal effects of hepatic metabolism only occur in the presence of a pituitary in situ.     

Neonatal estrogen treatment alters sexual differentiation of hepatic histidase.

Hepatic histidase was used as an enzyme marker for the study of neonatal programming in intact rats. Diethylstilbestrol (DES) or 17 beta-estradiol (E2) treatment for days, 2, 4, and 6 post partum resulted in decreased histidase activities in the adult female, but no effect was seen in prepubertal male and female rats or in adult males. In contrast, similar neonatal doses of testosterone propionate (TP) had no effect on histidase. Dose-response experiments demonstrate a 3-fold greater neonatal sensitivity to DES than to E2. The action of neonatal estrogen treatment is demonstrated to be permanent and irreversible. Neonatal treatment with E2, DES, or TP resulted in decreased uterine wet weights in adult females (E2 less than DES less than TP less than controls). Circulating sera estrogen levels were lower in adult E2- and DES-treated females than in TP-treated and control females. Our results suggest that these alterations may be due to direct toxic effects on the postnatal development of the female reproductive tract and endocrine system and/or to organizational effects on nerve endings in the hypothalamus that result in programming for altered sexual differentiation of hepatic metabolism.     

A simple chemosensory response in Drosophila and the isolation of acj mutants in which it is affected.

Although the Drosophila visual system has been described extensively, little is known about its olfactory system. A major reason for this discrepancy has been the lack of simple, reliable means of measuring response to airborne chemicals. This paper describes a jump response elicited by exposing Drosophila to chemical vapors. This behavior provides the basis for a single-fly chemosensory assay. The behavior exhibits dose dependence and chemical specificity: it is stimulated by exposure to ethyl acetate, benzaldehyde, and propionic acid but not ethanol. Animals can respond repeatedly at short intervals to ethyl acetate and propionic acid. The response relies on the third antennal segments. To illustrate the use of this behavior in genetic analysis of chemosensory response, nine acj mutants defective in response are isolated (acj = abnormal chemosensory jump), and their responses to two chemicals are characterized. All of the acj mutants are normal in giant fiber system physiology, and two exhibit defects in visual system physiology.     

Effect of estrogen plasma binding on sexual differentiation of the rat fetus.

alpha-Fetoprotein, the estradiol-binding plasma protein (EBP), binds estradiol but not R 2858 (11beta-methoxy-17-ethynyl-estradiol) specifically. R 2858 interferes more markedly than estradiol with the sexual differentiation of the male rat fetus following treatment of the mother during the final stages of gestation. Moreover, its tissular uptake is higher. These facts suggest that alpha-fetoprotein protects the fetus from the high circulating hormone concentrations present in the pregnant mother. The hormone, once transferred to the fetus, is retained in its vascular bed by EBP.     

�Ա�������Էֻ��쳣

表1影响性腺分化的有关基因及其突变后表型特征基因突变后表型特征WT1性腺发育不良 肾小球硬化,肾病,肾功能衰竭,伴发Wilms瘤风险,有间性性发育异常(Denys-Drash及Frasier综合征)SF1性腺发育不良,肾上腺功能低下DAX1基因复制:XY性腺发育不良基因突变:先天肾上腺发育不良(AHC)(X     

Epigenetic mechanisms are involved in sexual differentiation of the brain

Sexual differentiation of the brain can be considered as a process during which effects of sex steroid hormones secreted during early development is maintained into adulthood. Epigenetic regulation is emerging as a potentially important mechanism of conveyance of long-lasting effects of the hormonal and environmental milieu in the developing brain. Evidence has accumulated to show that epigenetic regulation is involved in the control of sexual differentiation of the brain. In the preoptic area (POA), which is important for male sexual behavior, histones associated with the estrogen receptor (ER) α and aromatase (Arom) gene promoters are differentially acetylated between the sexes, and two subtypes of histone deacetylase (HDAC2 and 4) are associated with the same promoters at higher frequencies in males in the early postnatal period. Since ERα and Arom are essential genes in masculinization of the brain, these findings suggest that histone deacetylation in the early postnatal period is involved in masculinization of the brain. Indeed, inhibition of HDAC activity in males during this period abrogates brain masculinization: structural sexual dimorphism of the bed nucleus of the stria terminalis is eliminated and expression of male sexual behavior is reduced in adulthood. Previous reports have demonstrated that ERα gene expression in the POA is higher in females during the developmental and pubertal periods and in adulthood, indicating that sexually dimorphic ERα expression that appears in early postnatal development is maintained until adulthood by epigenetic programming. The ERα promoter is also more sparsely methylated in females, with an inverse correlation with ERα expression. In addition to the hormonal effect, the amount of maternal care received during postnatal development has a lasting effect on ERα expression mediated by DNA methylation of its promoter. Taken together, these results suggest that epigenetic mechanisms play a central role in the transduction and maintenance of early hormonal and social cues to organize sexually differentiated brain functions.     

The amygdaloid complex in the sexual differentiation of the brain]

Presents the results of many-year research of the systemic and structural functional arrangement of the amygdaloid complex of the rat brain as the neuroendocrine center involved in the regulation of the reproductive processes. Special attention was paid to the sexual dependence of the structural and functional arrangement of this section of the brain. The amygdaloid complex was found involved in the critical period of brain development in the processes of sexual differentiation, this fact resulting in sexual dimorphism, that manifests in sexual differences of this complex structural and functional arrangement and its histophysiology. Sexual dimorphism areas in each of the neuronal ensembles of the rat amygdaloid complexes singled out by the authors, are described.     

Monensin-resistant LLC-PK1 cell mutants are affected in recycling of the adenylate cyclase-stimulating vasopressin V2-receptor.

The ionophore monensin was found to markedly reduce the rate of return of vasopressin V2-receptors to the membrane following down-regulation with [Arg8]vasopressin (AVP), as well as hormone dissociation (unloading) from cells following ligand binding and internalization in LLC-PK1 renal epithelial cells. Monensin-resistant LLC-PK1 mutants were isolated and characterized for V2-receptor recycling. Whilst the MN-41 mutant appeared to be impaired in [3H]AVP internalization, the MN-11 and MN-21 mutants exhibited parental V2-receptor binding and internalization, but markedly impaired receptor recycling subsequent to ligand-dependent receptor down-regulation. Unloading subsequent to ligand binding and internalization at 37 degrees C was also much slower in the mutants either at 37 degrees C or 23 degrees C. In contrast, unloading subsequent to binding at 23 degrees C, or to binding at 37 degrees C in the presence of NH4Cl, was comparable in LLC-PK1 and mutant cells implying the active nature of the recycling process impaired in the mutants. The mutations conferring resistance to monesin thus concomitantly impaired V2-receptor recycling in the mutants. Results argue for a monensin-sensitive endosomal/lysosomal pathway for the renal V2-receptor, representing the first such report for an adenylate cyclase stimulating receptor.     

Cytoduction in Chlamydomonas reinhardtii.

After conjugation between Chlamydomonas gametes of opposite mating type, a transient dikaryon is formed. The two nuclei fuse within 4-6 hr after mating. The young diploid zygote differentiates into dormant zygospore competent to complete meiosis, or more rarely (2-10% of cases) it undergoes mitosis to produce a stable diploid progeny. We here bring genetical, biochemical, and cytological evidence that among the mitotic zygotes, a large proportion of them undergo cytokinesis without fusion of the nuclei-a process that has been termed "cytoduction." By using appropriate genetic markers, haploid cytoductants that possess the nuclear genotype of one parent and the chloroplast marker of the other parent can easily be isolated. Genetical analysis and hybridization experiments moreover show that many haploid cytoductants transmit the chloroplast DNA molecules of both parents and that, as in diploids, these DNA copies occasionally recombine. This process of cytoduction extends the life cycle of Chlamydomonas and provides new tools for its genetic analysis.     

Pteridines as nonretinal regulators of light-dependent melatonin biosynthesis.

6-Formylpterin was purified by paper chromatography, and bovine pineal hydroxyindole-O-methyltransferase (S-adenosyl-L-methionine:N-acetylserotonin O-methyl-transferase; EC 2.1.1.4) was purified by flat-bed gel electrofocusing. Fractions of the methyltransferase with isoelectric points at pH 5.0, 5.7, and 5.2 were tested in the presence of 6-formylpterin, and decreased activities were found. Because 6-formylpterin is one of the photolytic products of folic acid and decreased serum folic acid has been reported in light-exposed men, we suggest that pteridines may be involved in nonretinal effects of light on pineal melatonin biosynthesis. It may be that this metabolic pathway is an evolutionary relic of primitive light responses of pigmentation mechanism.     

Correspondence between sexual isolation and allozyme differentiation: a test in the salamander Desmognathus ochrophaeus.

Ethological reproductive isolation and genetic divergence across 26 protein loci were measured among populations of the salamander Desmognathus ochrophaeus in the southern Appalachian Mountains. Levels of ethological isolation varied from none to complete and were statistically significant for all but two pairings between populations inhabiting different mountain ranges. When geographic and genetic distances were treated as independent variables in multiple correlation analyses, they accounted for about half the variance in levels of ethological isolation. When genetic distance is held constant, the remaining relationship between ethological isolation and geographic distance is still statistically significant. When geographic distance is held constant, the remaining relationship between genetic distance and levels of ethological isolation is nonsignificant, as is the relationship between geographic distance and genetic distance when ethological isolation is held constant. Ethological isolation and genetic divergence evidently both reflect the gradual divergence of allopatric populations, but genetic distance is a poor predictor of ethological isolation in these salamanders.     

Ectopic germ cells: natural model for the study of germ cell sexual differentiation.

In the course of a study on the morphogenesis of the adrenal gland in random-bred Swiss albino mice, we noted the presence of ectopic germ cells in the adrenal cortexes and medullas in animals of both sexes, from day 12 1/2 of fetal development to postnatal day 12. Up to day 15 of fetal development, the cells exhibited characteristics of primordial germ cells. At day 17, and irrespective of the sex of the fetus, they all entered meiosis in synchrony with those in the ovary. Postnatally, in females as well as males, all ectopic germ cells displayed morphologic characteristics identical to those of young oocytes in unilaminar ovarian follicles. No germinal elements were seen in the adrenal glands past day 12 of life. Our study shows that mammalian germ cells are capable of undergoing sustained differentiation outside the gonads and that, in ectopic sites, they all differentiate into oocytes as they normally would in the ovary, even in males.