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E S Robinson

Publications and source records attributed to E S Robinson.

At least 55 records · Page 3Linked to original sources

X chromosome inactivation in female embryos of a marsupial mouse (Antechinus stuartii).

Blastocysts and late gestation stages of the marsupial mouse, Antechinus stuartii, were examined cytologically and electrophoretically to investigate X chromosome activity during embryogenesis. A late replicating X chromosome was identified in the protoderm cells of female unilaminar blastocysts and in the cells of embryonic and extra-embryonic regions of older blastocysts. Sex chromatin bodies were also observed in female bilaminar and trilaminar blastocysts. The X linked enzyme alpha-galactosidase showed no evidence of paternal allele expression in the extra-embryonic region of bilaminar blastocysts or in the yolk sac and embryonic tissue of known heterozygotes. It is concluded that the late replicating X chromosome is paternal in origin and that unlike the laboratory mouse, X inactivation is not correlated with cell differentiation in Antechinus.

Animals↗

X-linked gene expression and X-chromosome inactivation: marsupials, mouse, and man compared.

The existence of paternal X inactivation in Australian and American marsupial species suggests that this feature of X-chromosome dosage compensation is not a recent adaptation, but probably predates the evolutionary separation of the Australian and American marsupial lineages. Although it is theoretically possible that the marsupial system is one of random X inactivation with p greater than 0.99 and q less than 0.01 and dependent on parental source, no instance of random X inactivation (p = q or p not equal to q) has ever been verified in any tissue or cell type of any marsupial species. Therefore, we conclude that the most fundamental difference in X inactivation of marsupials and eutherians is whether the inactive X is the paternal one or is determined at random (with p = q in most but not all cases). The only other unequivocal difference between eutherians and marsupials is that both X chromosomes are active in mice and human oocytes, but not in kangaroo oocytes. Apparently, the inactive X is reactivated at a later meiotic stage or during early embryogenesis in kangaroos. X-chromosome inactivation takes place early in embryogenesis of eutherians and marsupials. Extraembryonic membranes of mice exhibit paternal X inactivation, whereas those of humans seem to exhibit random X inactivation with p greater than q (i.e., preferential paternal X inactivation). In general, extraembryonic membranes of marsupial exhibit paternal X inactivation, but the Gpd locus is active on both X chromosomes in at least some cells of kangaroo yolk sac. It is difficult to draw any general conclusion because of major differences in embryogeny of mice, humans, and marsupials, and uncertainties in interpreting the data from humans. Other differences between marsupials and eutherians in patterns of X-linked gene expression and X-chromosome inactivation seem to be quantitative rather than qualitative. Partial expression of some genes on the inactive X is characteristic of marsupials, with species variation in the behavior of specific loci; some X-linked human genes on the inactive chromosome also are known to exhibit partial activity in vivo and in cultured cells. The X chromosomes of marsupials do not behave as units with respect to transcriptional activity, nor does the human X chromosome. In addition, Barr bodies have recently been detected at interphase in some marsupials, establishing that this manifestation of X chromosome inactivity is not restricted to eutherians.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Lack of correlation between Gpd expression and X chromosome late replication in cultured fibroblasts of the kangaroo Macropus robustus.

Cultured fibroblasts and lymphocytes from M. robustus females heterozygous for the X-linked Gpd gene were examined electrophoretically and cytologically. Gpd expression in lymphocytes was restricted to the maternal allele while in fibroblasts there was also partial expression of the paternal allele. The Gpd gene is thought to be located on the long arm of the X chromosome. However, in fibroblasts the long arm of the paternal X chromosome showed no indication of an early replicating segment.

Animals↗

Influence of dietary tryptophan on the induction of gamma-glutamyltranspeptidase-positive foci in the livers of rats treated with hepatocarcinogen.

The ingestion of an elevated level (2%) of L-tryptophan (TRP) in a purified diet was investigated to determine whether it would influence the induction of gamma-glutamyltranspeptidase (GGT)-positive foci in the livers of rats exposed to a hepatocarcinogen. Subtotal hepatectomies were performed, and 18 h later, the rats were given injections i.p. of diethylnitrosamine (30 mg/kg). Ten days later, groups of male rats were placed on choline-supplemented (CS), CS + TRP, choline-deficient (CD), or CD + TRP diets for 10 wk. In two separate experiments, the rats fed the CS + TRP diet or the CD diet developed more and larger GGT + foci than did rats fed the CS diet. Rats fed the CD + TRP diet revealed similar changes to those found in rats fed the CD diet. The liver weights of the rats fed the CD or the CD + TRP diet were greater than those of rats fed the CS or the CS + TRP diet. Hepatic GGT activity was somewhat elevated in rats fed the CS + TRP diet and markedly elevated in rats fed the CD or the CD + TRP diet. Hepatic ornithine decarboxylase activity was increased in rats fed the CD + TRP diet. The results suggest that increased dietary tryptophan has a promoting effect on liver carcinogenesis as measured by the induction of GGT + foci in the livers of rats exposed to diethylnitrosamine. A potentiating effect by tryptophan was not observed in the livers of rats fed a CD diet.

Amino Acids↗

Dictyate oocytes of a kangaroo (Macropus robustus) show paternal inactivation at the X-linked Gpd locus.

Purified samples of large numbers of dictyate oocytes from 13 M. robustus pouch young heterozygous for glucose-6-phosphate dehydrogenase type and six homozygous controls were examined electrophoretically to determine activity states at the Gpd locus. Like somatic cortical and medullary cells, oocytes expressed only the maternal phenotype irrespective of the direction of the cross. No evidence was found of reactivation of the inactive (paternal) allele or inactivation of both maternal and paternal alleles. It was therefore concluded that unlike eutherian dictyate oocytes, only a single (maternal) allele is active in each dictyate oocyte in M. robustus. The stage of reactivation of the paternal allele remains to be determined.

Animals↗

Glucose-6-phosphate dehydrogenase and lactate dehydrogenase activity in kangaroo and mouse oocytes.

Glucose-6-phosphate dehydrogenase and lactate dehydrogenase activity in oocytes and follicle cells of the Australian marsupial, Macropus eugenii (the tammar), and the mouse were determined by a simplified microelectrophoresis method. Mouse oocytes have approx 285 times more G6PD activity per picolitre of cytoplasm than tammar oocytes and about 10 times more LDH. The ratio of LDH to G6PD in mouse follicle cells is close to 3 whereas in the tammar it approaches unity. The very low levels of activity of G6PD in tammar oocytes may be due to transcriptional, translational or metabolic differences compared with the mouse.

Animals↗

Germ cell development in female pouch young of the tammar wallaby (Macropus eugenii).

The cytology, timing, numbers and arrangement of germ cells in gonads of 35 female tammar wallaby pouch young aged from newborn to 210 days post partum were determined from serial sections. Germ cell cytology was essentially similar to that in eutherians. The total number of germ cells per ovary reached a peak of about 4.5 X 10(5) at 50 days and was reduced to about 25% of the maximum value by 210 days. The first oogonia developed from primordial germ cells soon after birth; the first meiotic oocytes were formed between 24 and 30 days; follicularization of diplotene oocytes began at about 50 days, and by 210 days normal, prefollicular germ cell development had ended. Successive stages of germ cells were initiated at the inner margin of the cortical region of the gonad and spread as a series of waves to the tunica albuginea. At all stages of oogonial and oocyte development many cells showed characteristic degenerative changes leading to atresia. The timing of female germ cell development in relation to gestation in tammars and humans is comparable if tammar gestation is taken to include pouch life.

Aging↗

X-chromosome inactivation and evolution in marsupials and other mammals.

Sex chromosome dosage compensation in Australian marsupials exhibits several characteristics not ordinarily manifested in eutherian mammals. They are: 1) preferential inactivation of the paternally derived X-chromosome in somatic tissues; 2) a state of partial inactivity at some paternally derived X-linked gene loci in some cells; 3) absence of dosage compensation in some cell types; 4) absence of nuclear sex dimorphism at interphase; 5) discordance between late DNA replication and gene inactivation in some cell types; 6) failure of the X-chromosome to behave as a single unit. Despite these seemingly major differences between the dosage compensation systems of Australian marsupials and eutherians, there are also many underlying similarities. We propose that the two systems arose from a common ancestral mechanism in which the X-chromosome was regulated in a piecemeal fashion, rather than as a whole. To determine more precisely the sequence and timing of evolutionary events, we have initiated an investigation of dosage compensation in American marsupials, which diverged from Australian marsupials soon after the divergence of marsupials from eutherian mammals.

Animals↗

Polyploidy in the australian leptodactylid frog genus Neobatrachus.

Karyotypic analysis of six species of the Australian leptodactylid frog genus Neobatrachus showed that N. pictus, N. centralis, N. pelobatoides and N. wilsmorei are diploid (2n = 24) while N. sudelli and N. sutor are tetraploid (4n = 48). Polyploidy has not been reported previously among Australian anurans. Idiograms of the six species indicate that they are similar to the other Australian leptodactylids so far discribed. DNA values of the tetraploids are approximately double the values for diploids. Tetraploid nuclear and cell sizes are greater compared with diploids but total body size shows no increase. At diakinesis in primary spermatocytes of tetraploids, mainly tetravalents together with a few bivalents are present. Silver straining of metaphase spreads clearly demonstrates the location of NORs at the secondary constrictions and their frequent association in the tetraploid N. sutor. Nucleolar number in interphase nuclei provides a reliable guide for distinguishing tetraploid from diploid frogs in the absence of chromosome analysis and can be determined for both living and preserved specimens. The possible origins and relationships of the tetraploid species are discussed.

Animals↗

Acute stress and the brain norepinephrine uptake mechanism in the rat.

The kinetic constants for norepinephrine uptake in cerebral cortical homogenates were determined in vitro immediately following an acute stress consisting of either forced immobilization, cold-wet exposure, combined cold-plus-restraint, swim stress, or electric footshock in the rat. The kinetic constants, apparent Km and Vmax, for uptake of 3H-l-norepinephrine were significantly increased only following 10 min swim at 22 degrees or following 5 min electric footshock. When severe hypothermia accompanied the stress, the findings suggested that a profound reduction in body temperature was associated with depressed responsiveness of brain noradrenergic mechanisms to stress including decreased uptake kinetic constants. In a series in which the duration of electric footshock was varied from 2 to 30 min, it was noted that the NE uptake kinetic constants were increased at 5 min, but were similar to paired controls at 2, 10 and 30 min following the onset of footshock. It was concluded that various acute stresses did not elicit a generalized response of the cortical NE uptake mechanism to stress in the rat. Furthermore, when uptake kinetic constants did change with stress, the values were often within the range of normal values seen in the rat.

Animals↗