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PubMedDate not supplied

A novel single nucleotide polymorphism in the 3' untranslated region of human glutathione peroxidase 4 influences lipoxygenase metabolism.

Selenium (Se) is an essential micronutrient for human health. The biological roles of the essential micronutrient Se are attributed to its presence in a range of 20-30 selenoproteins including the cytosolic and phospholipid hydroperoxide glutathione peroxidases (GPX1 and GPX4). It has been suggested that GPX4 may play a role in regulation of leukotriene biosynthesis and thus inflammation. In eukaryotes Se is incorporated into selenoproteins as the amino acid selenocysteine in a process requiring a stem-loop within the 3' untranslated region (3'UTR) of the mRNA. In this study the region of the GPX4 gene corresponding to the 3'UTR was scanned for mutations in a group of 66 volunteers. The data show a T/C variant at position 718. The distribution of this SNP in our population was 34% CC, 25% TT and 41% TC; i.e., it is in Hardy-Weinberg equilibrium. Individuals of different genotypes exhibited significant differences in the levels of lymphocyte 5-lipoxygenase total products, with C718 showing increased levels of those products compared to T718 and T/C718 (36% and 44% increases, respectively). The data suggest that the SNP718 that we have identified has functional effects and support the hypothesis that GPX4 plays a regulatory role in leukotriene biosynthesis.

3' Untranslated Regions↗
PubMed1995

Genetic regulation of shoot development in Arabidopsis: role of the EMF genes.

To investigate the genetic mechanism regulating Arabidopsis shoot maturation and development, we characterized eight emf mutants that bypassed the vegetative phase of the life cycle. Genetic complementation studies identified two EMF loci; both mapped to chromosome five. Double mutant analysis showed that the early- and late-flowering mutants, co, fb, elf1, elf2, and elf3, could not rescue vegetative development in the emf mutants, confirming the need for both EMF gene activities for rosette development. A series of phenotypes involving successive loss of reproductive organs was also observed in emf single mutants, in emf1-1/emf1-2 transheterozygotes, and in emf1 emf2 double mutants, suggesting that the EMF genes not only specify the rosette (vegetative) but also are involved in inflorescence and flower (reproductive) development. Phenotypic analysis of double mutants between emf and tfl1, lfy, and ag indicated interactions between EMF and genes regulating inflorescence meristem development and floral organ identity. A model depicting the role of the EMF genes in regulating shoot maturation and their interaction with genes that affect phase transitions is presented.

Alleles↗
PubMed1999

NT4/5 mutant mice have deficiency in gustatory papillae and taste bud formation.

Neurotrophins are key determinants for controlling the survival of peripheral neurons during development. Brain-derived neurotrophic factor (BDNF) and neurotrophin-4/5 (NT4/5) exert their action through a common trkB receptor but independently support gustatory sensory neurons. To assess the role of NT4/5 during development, we examined the postnatal development and maintenance of fungiform taste buds in mice carrying a deletion of NT4/5. The absence of NT4/5 results in embryonic deficits in gustatory innervation and a reduced number of fungiform papillae at birth. No degenerative deficits of fungiform papillae were observed for the first 3 weeks of postnatal development. However, these remaining fungiform papillae were smaller in appearance and many did not contain taste pores. By postnatal day 60, there was 63% decrease in the number of fungiform papillae, and remaining papillae were smaller in size or modified into filiform-like spines. These papillae had either no taste bud or a taste bud with a reduced number of taste cells compared to controls. These findings demonstrate that the NT4/5 gene functions in the maintenance of fungiform gustatory papillae and raises the possibility for an earlier role in development.

Animals↗
PubMed1997

Involvement of heparan sulfate proteoglycans in the binding step for phagocytosis of latex beads by Chinese hamster ovary cells.

Chinese hamster ovary (CHO) K1 cells, typical nonprofessional phagocytes, exhibited intense phagocytosis of latex beads when incubated under serum-free conditions. Under the serum-free conditions, the recognition mechanism of latex beads by cells was investigated. Exogenous heparin and heparan sulfate but not chondroitin sulfate effectively inhibited the binding of latex beads to cells. The binding of latex beads to cells was also inhibited by treatment of cells with heparitinase more effectively than by treatment of cells with chondroitinase. Furthermore, CHO mutant cells defective in biosyntheses of both heparan sulfate and chondroitin sulfate proteoglycans almost completely lacked binding activity of latex beads. Another mutant, which is deficient in heparan sulfate proteoglycans but rather overproduces chondroitin sulfate proteoglycans, also showed lower binding activity, compared with wild-type cells. Coculture of these proteoglycan-less mutants and the wild-type cells did not restore the binding activity of the mutant cells, suggesting that membrane-bound rather than secretory proteoglycans were responsible for the binding of latex beads. These results indicated that heparan sulfate proteoglycans at the cell surface were involved in the binding step for phagocytosis of latex beads by CHO cells.

Animals↗
PubMed1997

Widespread neuronal ectopia associated with secondary defects in cerebrocortical chondroitin sulfate proteoglycans and basal lamina in MARCKS-deficient mice.

Mice deficient in MARCKS, a prominent neural substrate for protein kinase C (PKC), die before or shortly after birth. They exhibit high frequencies of exencephaly, universal agenesis of forebrain commissures, and abnormalities of cerebral cortical and retinal lamination. We show here that these mice have wide-spread and severe neuronal ectopia in the outer layers of the developing forebrain, manifested by the migration of clusters of developing neuroblasts through the basal lamina and often through the pial membrane and into the subarachnoid space. This abnormality became apparent by Embryonic Day (E) 13 or 14, shortly after the formation of the early marginal zone. MARCKS deficiency was associated with decreased staining for marginal zone chondroitin sulfate proteoglycans; this decrease was detectable earlier in development than the neuronal ectopia. Later in development, there was also marked disruption of the basal lamina at the pial-glial interface, as evidenced by gross abnormalities in laminin and reticulin staining; however, the basal lamina appeared normal at E9.5. These data indicate that MARCKS is required for the prevention of neuronal ectopia during development. Potential mechanisms responsible for the neuronal ectopia in the MARCKS-deficient mice include decreased expression or increased proteolytic destruction of basal lamina proteins and marginal zone chondroitin sulfate proteoglycans in the developing brain.

Animals↗
PubMed1999

Microconidia of Neurospora crassa.

Neurospora crassa produces two types of vegetative spores-relatively small numbers of uninucleate microconidia and very large numbers of multinucleate macroconidia (blastoconidia and arthroconidia). The microconidia can function either as spermatia (male gametes) or as asexual reproductive structures or both. In nature they probably function exclusively in fertilization of protoperithecia. The environmental conditions favoring their formation and the pattern of their development are quite distinct from those of macroconidia. Mutants of N. crassa have been isolated in which macroconidiation is selectively blocked without affecting microconidiation, showing that these two types of conidial differentiation involve distinct developmental pathways. Unlike microconidia of some related ascomycetes, those of Neurospora are capable of germination, providing viable uninucleate haploid cells which are desired in several types of investigations. A technique of selectively removing macroconidia from culture initiated on cellophane overlying agar medium allows pure microconidia to be obtained even from the wild-type strains of Neurospora. The conditional microcyclic strain, mcm, allows either macroconidia or microconidia to be obtained at will, depending on the conditions of culture. The new methods of obtaining pure microconidia from normal laboratory strains will make it quick and easy to purify heterokaryotic transformants following introduction of DNA into multinucleate protoplasts. Moreover, these methods allow the detection of genetic variability that remains hidden within an individual fungus and the estimation of the frequency of nuclear types in laboratory-constructed heterokaryons. The discovery, function, and development of microconidia are described and their research applications are discussed in this review.

Culture Media↗
PubMed2000

Normal phototransduction in Drosophila photoreceptors lacking an InsP(3) receptor gene.

The Drosophila light-sensitive channels TRP and TRPL are prototypical members of an ion channel family responsible for a variety of receptor-mediated Ca(2+) influx phenomena, including store-operated calcium influx. While phospholipase Cbeta is essential, downstream events leading to TRP and TRPL activation remain unclear. We investigated the role of the InsP(3) receptor (InsP(3)R) by generating mosaic eyes homozygous for a deficiency of the only known InsP(3)R gene in Drosophila. Absence of gene product was confirmed by RT-PCR, Western analysis, and immunocytochemistry. Mutant photoreceptors underwent late onset retinal degeneration; however, whole-cell recordings from young flies demonstrated that phototransduction was unaffected, quantum bumps, macroscopic responses in the presence and absence of external Ca(2+), light adaptation, and Ca(2+) release from internal stores all being normal. Using the specific TRP channel blocker La(3+) we demonstrated that both TRP and TRPL channel functions were unaffected. These results indicate that InsP(3)R-mediated store depletion does not underlie TRP and TRPL activation in Drosophila photoreceptors.

Animals↗
PubMed1998

Molecular analysis and prenatal diagnosis of human fumarase deficiency.

Fumarase deficiency is a rare autosomal recessive disorder of the citric acid cycle causing severe neurological impairment. The cDNA for both the rat and human enzymes has been cloned previously and shown to encode a coding region of 1.46 kb. To scan for mutations in fumarase-deficient patients we amplified the coding region of fumarase from fibroblast/lymphoblast cDNA employing the oligonucleotide primers designed from the published human and rat cDNA sequence. We then directly sequenced the polymerase chain reaction product. In seven unrelated patients, we detected four missense mutations (A265T, D383V, F269C, K187R), a nonsense mutation (W458X), a 3-bp AAA insertion that introduces an additional lysine residue at codon 435, and a spontaneous new mutation resulting in a 74-bp deletion (66del74). Seven at-risk pregnancies were monitored with one prenatal diagnosis of fumarase deficiency by molecular analysis and favorable outcome of the other pregnancies as predicted by enzyme assay of cultured fetal cells or molecular analysis.

Amino Acid Metabolism, Inborn Errors↗
PubMed1999

The effect of the pneumococcal toxin, pneumolysin on brain ependymal cilia.

Densely ciliated ependymal cells cover the ventricular surface of the brain and cerebral aqueducts separating cerebrospinal fluid, which is infected in meningitis, from neuronal tissue. We have established an ex vivo model that allows measurement of ependymal ciliary beat frequency, using high-speed video analysis, during incubation with bacterial toxins. Ciliated ependyma, from Wistar rats, was exposed to the pneumococcal toxin, pneumolysin, and a mutant form with markedly reduced cytotoxic activity (;0.1%). Wild-type pneumolysin (1500 HU/ml and 150 HU/ml: 10 and 1 microg/ml) caused rapid ciliary stasis (30-150 s), sloughing of cilia and cytoplasmic extrusion. Ciliary slowing before stasis was seen at 15 HU/ml (0.1 microg/ml); however, no effect on ciliary beat frequency was seen at lower concentrations (1.5 HU/ml and 0.15 HU/ml: 0.01 and 0.001 microg/ml). Mutant pneumolysin, 99.9% deficient in haemolytic activity, caused rapid ciliary stasis at 10 microg/ml but no effect was seen at lower concentrations (1-0.1 microg/ml). Pneumolysin, at levels which may be produced during severe pneumococcal meningitis, may cause rapid ependymal ciliary stasis.

Animals↗
PubMed1991

Nerve cells of Drosophila Notch mutant are differentiated inside amphibian brain: a new approach for the analysis of genetic control of nerve cell differentiation.

Fragments of the neural primordium of a new Notch mutant of Drosophila melanogaster produced in our laboratory were transplanted into the neural tube of embryos of 4 amphibian species (caudate and ecaudate) immediately after completion of neurulation. The grafts were identified by using a light microscope, scanning electron miscroscope, and in situ hybridization with mobile genetic elements of Drosophila and fluorescent dyes as markers. As has been shown, Drosophila nerve cells survive and differentiate inside the neural tube of amphibian embryos. The grafts increase in size by twentyfold and the cell proliferation zones are retained during the period of six months. Differentiated cells of the graft formed axon-dendritic contacts with recipient cells and penetrated into the organisms' brain structures. The effect of Drosophila transplants proved to be different for caudate and ecaudate amphibians. The presence of the graft accelerated the development of Xenopus laevis and it also affected their behavior. This approach can be very useful for the study of genetic basis of development and behavior.

Animals↗
PubMed1979

An SEM AND TEM study of suppression of eye development in eyeless mutant axolotls.

Morphology of the primary optic rudiment of normal eyed and mutant eyeless (e/e) axolotl embryos was studied at light. Tem and SEM levels. The presumptive eyeforming region of eyeless embryos differs from that of normal embryos in several important respects including premature formation of basal lamina, separation from overlying ectoderm by mesenchyme cells and persistence of granules in the interspace surrounding the optic anlage into relatively late developmental stages. These differences suggest that the gene that causes failure of eye formation in the mutant axolotls produces structural differences that interfer with normal physical and/or biochemical inductive interactions between neurectoderm and mesenchyme cells due primarily to the precocious development of basal lamina over the extermal surface of the optic primordia.

Ambystoma↗
PubMed1986

Inhibition of wall autolysis of staphylococci by sodium polyanethole sulfonate "liquoid".

Liquoid (polyanethole sulfonate) was neither capable of influencing the growth nor the viability of staphylococci. But liquoid induced a suppression of the activity of different autolytic wall systems of normally growing staphylococci, i.e., autolysins which participate in cross wall separation as well as autolysins which are responsible for cell wall turnover. Additionally, the lysostaphin-induced wall disintegration of staphylococci was inhibited by liquoid. However, no indication could be found for a direct inhibition of lytic wall enzymes by liquoid; rather an interaction of liquoid with the target structure for the autolytic wall enzymes, the cell wall itself, was postulated. On the basis of the experimental data with the teichoic acid- mutant S. aureus 52A5 the sites of wall teichoic acid were supposed to be an important target for the binding of liquoid to the staphylococcal cell wall.

Bacteriolysis↗
PubMed1991

Ultrastructure and cell wall composition in cell division cycle mutants of Schizosaccharomyces pombe deficient in septum formation.

A number of temperature-sensitive cdc- mutants of Schizosaccharomyces pombe that are affected in septum formation were analyzed with respect to their ultrastructure and the composition of their cell wall polymers. One mutant strain, cdc 16-116, has a cell wall composition similar to the wild type (strain 972 h-). However two other mutants, cdc 4 and cdc 7, show a higher galactomannan content and a lower alpha-glucan content. In all the mutants tested, total glucose incorporation, protein, RNA and DNA synthesis increased similarly to wild type over 3 1/2 h. After 2-3 h of incubation at the non permissive temperature -35 degrees C-, cell numbers remained constant although, increases in optical densities at 600 nm were observed. According to scanning electron microscopy, the mutants had aberrant shapes after 5 h of incubation at 35 degrees C. Transmission electron microscopy showed that cdc 3 is unable to complete septum formation. cdc 4 showed the most varied morphological shapes and aberrant depositions of cell wall material. cdc 8 exhibited a deranged plasma membrane and cell wall regions near of cell poles; an abnormal septum and several nuclei. cdc 7 showed elongated cells with several nuclei and with an apparently normal cell wall completely lacking in septum and septal material. cdc 16 showed more than one septum per cell.

Cell Cycle↗