Formation of two microtubule-nucleating sites which perform differently during centrosomal reorganization in a mouse cochlear epithelial cell.

This report provides evidence for the formation of a cell surface-associated centrosome with two spatially discrete microtubule-nucleating sites that perform differently; the minus ends of microtubules remain anchored to one site but escape from the other. Centrosomal reorganization in the cells in question, outer pillar cells of the organ of Corti, indicates that its pericentriolar material becomes intimately associated with the plasma membrane at the two nucleating sites. Two large microtubules bundles assemble in each cell.

The effects of calyculin A upon calcium-, guanine nucleotides- and phorbol 12-myristate 13-acetate-stimulated ACTH secretion from AtT-20 cells.

1. The mouse AtT-20/D16-16 anterior pituitary tumour cell line was used as a model system for the study of protein phosphatase involvement in the late stages of the secretory pathway for adrenocorticotrophin (ACTH) secretion. The effects of the type 1 and 2 phosphatase inhibitor calyculin A upon calcium-, guanine nucleotide- and phorbol 12-myristate 13-acetate (PMA)-stimulated ACTH secretion from electrically-permeabilized AtT-20 cells were studied.

Studies on a stabilisation of ubisemiquinone by Escherichia coli quinol oxidase, cytochrome bo.

The Escherichia coli quinol oxidase, cytochrome bo, is closely related to the cytochrome c oxidase, cytochrome aa3 in all aspects of its structure and function except for the replacement of the cytochrome-c-binding site and its attendant CuA prosthetic group with a quinone-binding site. The putative oxidation of quinol by ferrihaem (cytochrome b) at this site in sequential one-electron steps requires the stabilisation of semiquinone. We have observed, by electron paramagnetic resonance, the properties of a ubisemiquinone radical in appropriately poised samples of purified enzyme reconstituted with excess ubiquinone.

Involvement of brainstem serotonergic interneurons in the development of a vertebrate spinal locomotor circuit.

Brainstem neurons modulate the rhythmic output of spinal locomotor circuitry in adult vertebrates, but how these influences develop is largely unknown. We demonstrate that the ingrowth of serotonergic axons to the spinal cord of Xenopus tadpoles plays a critical role in locomotor burst development by transforming the output of embryonic amphibian swimming circuitry into a more mature and flexible form. Our experiments show that exposure to a monoamine neurotoxin (5,7 dihydroxytryptamine) deletes serotonergic raphespinal projections and prevents the normal maturation of larval swimming.

Muscarinic agonists modulate calcium-dependent outward currents in an identified insect motoneurone.

The soma membrane of the fast coxal depressor (Df) motoneurone of the cockroach, Periplaneta americana possesses a population of acetylcholine receptors which respond to both nicotinic and muscarinic ligands. Activation of these 'mixed' acetylcholine receptors by McN-A-343 or oxotremorine results in the generation of an inward current at potentials positive to - 40 mV. A large proportion of the outward current induced by membrane depolarization in Df somata is due to a calcium-activated potassium conductance (IKCa) which shows a characteristic N-shape voltage-dependence.

Low genetic diversity in the Scottish endemic Primula scotica Hook.

Primula scotica Hook., a rare Scottish endemic, is threatened with extinction due to habitat destruction. A survey of genetic variability within the species was conducted on five populations from the Orkney islands and nine from the Scottish mainland.

Three microtubule-organizing centres collaborate in a mouse cochlear epithelial cell during supracellularly coordinated control of microtubule positioning.

Large cell surface-associated microtubule bundles that include about 3,000 microtubules assemble in certain epithelial cells called inner pillar cells in the mouse organ of Corti. Microtubule-organizing centres (MTOCs) at both ends and near the middle of each cell act in concert during control of microtubule positioning. In addition, the three cell surface-associated microtubule-organizing centres are involved in coordinating the connection of bundle microtubules to cytoskeletal components in neighbouring cells and to a basement membrane.

Properties of an identified dopamine-containing neurone in culture from the snail Helisoma.

The giant neurone in the left pedal ganglion of the snail Helisoma trivolvis is homologous with the giant dopaminergic neurone of Planorbis corneus, because the neurones have a very similar location and morphology, and react similarly with glyoxylic acid to produce an intense blue fluorescence, indicating the presence of dopamine. Each of these neurones is therefore referred to as a giant dopaminergic neurone, or GDN. Conditions for the extension of neurites and formation of chemical junctions in culture have been determined for the H.

NO, nitrosonium ions, nitroxide ions, nitrosothiols and iron-nitrosyls in biology: a chemist's perspective.

The multiplicity of biological functions thus far attributed to NO has led to suggestions that some effects might be mediated by other, related species instead. The radical nature of NO cannot account for its cytotoxicity, but its reaction with superoxide to form peroxynitite and highly reactive hydroxyl radicals may be important in this context. The ease with which NO can react with and destroy Fe-S clusters is also an important factor.

Spontaneous bursting induced by convulsant agents in an identified insect neurone.

1. Using an in vitro preparation, intracellular recordings were made from the cell body of the "fast" coxal depressor motoneurone (Df) from the cockroach, Periplaneta americana. 2.

A role for Y-box proteins in cell proliferation.

Members of the Y-box (YB) family of transcription factors are expressed in a wide range of cell types and are implicated in the regulation of a rapidly increasing number of genes. Although the biological activities of YB proteins appear to be varied, distinct patterns, relating to the timing of their expression and the identity of their target genes, are beginning to emerge. A recent report by Ito et al.

Binding of Y-box proteins to RNA: involvement of different protein domains.

Eukaryotic Y-box proteins are reported to interact with a wide variety of nucleic acid structures to act as transcription factors and mRNA masking proteins. The modular structure of Y-box proteins includes a highly conserved N-terminal cold-shock domain (CSD, equivalent to the bacterial cold-shock proteins) plus four basic C-terminal domains containing arginine clusters and aromatic residues. In addition, the basic domains are separated by acidic regions which contain several potential sites for serine/threonine phosphorylation.

Evidence that the paramyxovirus simian virus 5 can establish quiescent infections by remaining inactive in cytoplasmic inclusion bodies.

Following infection of BALB/c fibroblastic (BF) cells with simian virus 5 (SV5) only low levels of infectious virus were produced and the majority of cells survived the infection. However at 1 day post-infection (p.i.

Adenovirus protein-protein interactions: hexon and protein VI.

A variety of mastadenoviruses were denatured, their polypeptides separated by electrophoresis on SDS-polyacrylamide gels and transferred to nitrocellulose. The immobilized polypeptides were washed, incubated with buffers containing hexons from human adenoviruses (Ad) types 2, 5 and 12 and the location of bound hexons was detected with anti-hexon antibodies. It was found that hexons from any of the three human adenovirus types bound to protein VI from all the mastadenoviruses examined.

Enhanced chromosomal response of ataxia-telangiectasia cells to specific types of DNA double-strand breaks.

The chromosomal response of two ataxia-telangiectasia (A-T) lymphoblastoid cell lines (A-T-PA and A-T-KM) to restriction endonucleases (RE) is compared with that of a normal (N-SW) lymphoblastoid cell line. The RE used were PvuII (generating DNA double-strand breaks with blunt termini), BamHI (cohesive termini with 4 base, 5' overhangs) and PstI (cohesive termini with 4 base 3' overhangs). Chromatid aberrations were analysed in cells 5 h after treatment.

Glucocorticoid-induced changes in liver: effect of dexamethasone administration on DNA topoisomerase I and II activities and distribution of histone H1 subtypes.

The activities of DNA topoisomerase I and II and the relative proportions of the histone H1 subtypes were investigated in rat liver which was undergoing hypertrophy and exhibiting increased transcriptional activity following the administration of dexamethasone. There was a rise in the level of activity of DNA topoisomerase I and a slight fall in that of DNA topoisomerase II. The relative proportions of the H1 subtypes were altered due to a preferential increase in H1.

Radiation-induced transformation of SV40-immortalized human thyroid epithelial cells by single and fractionated exposure to gamma-irradiation in vitro.

Radiation-induced transformation of a human thyroid epithelial cell line (HTori-3) has been investigated following exposure to single and fractionated doses of gamma-irradiation. The human epithelial cells were irradiated in vitro and following passaging, transplanted to the athymic nude mouse. Following a single exposure to gamma-irradiation in the range 0.

Neural development. Chemoattractants for navigating axons.

Newly identified proteins that seem to act as diffusible attractants for circumferentially growing axons in the vertebrate embryonic spinal cord are related to a protein that directs circumferential axon growth in the nematode.

N-terminally extended FMRFamide-related peptides of Helix aspersa: processing of the precursor protein and distribution of the released peptides.

Sequencing of cDNA clones reveals a precursor protein that can be processed into 10 different hepta-FaRPs. Two of the peptides are previously undescribed and are N-terminally extended forms of-YMRFamide, making them the only methionine-containing peptides in the precursor. They are separated from the main cluster of hepta-FaRPs by a recognition site (RQKR) for the Golgi-resident proteolytic enzyme furin.

Neurons from mouse embryos with a null mutation in the tumour suppressor gene p53 undergo normal cell death in the absence of neurotrophins.

Cell death plays an important role in regulating cell numbers in a wide variety of tissues during development and throughout life. Cell death can be triggered by changes in the levels of hormones and growth factors and is regulated by the expression of the tumour suppressor gene p53 in many cells. To determine if p53 plays a role in neuronal death resulting from neurotrophin deprivation, we studied the survival of neurons obtained from normal mouse embryos and embryos with a null mutation in the p53 gene.

Phorbol ester activation of protein kinase C inhibits CNP-stimulated cyclic GMP production in the mouse AtT-20 pituitary tumour cell line.

Preincubation of AtT-20 mouse pituitary tumour cells with the phorbol ester PMA resulted in a concentration-dependent inhibition of CNP-stimulated cyclic GMP production. The phorbol ester analogue 4 alpha phorbol had no inhibitory effect and 24 h preincubations with PMA resulted in a characteristic down-regulation of the response indicating that the inhibitory actions were mediated via the activation of protein kinase C. Forskolin in the presence of the phosphodiesterase inhibitor IBMX stimulated intracellular cyclic AMP concentrations by up to eight fold, but did not alter basal nor CNP-stimulated cyclic GMP production.