Heteromers of Glutamate Decarboxylase Isoforms Occur in Rat Cerebellum

Abstract: The subunit structure of brain glutamate decarboxylase in cerebellum was investigated by using gel electrophoresis and antisera that specifically recognize the individual isoforms of brain glutamate decarboxylase (termed GAD₆₅ and GAD₆₇). The antisera were prepared against peptides that corresponded to amino acid sequences specific to each isoform. Each antiserum reacted specifically with the appropriate peptide in an ELISA and with the appropriate form of GAD on immunoblots. Nondenaturing gradient gel electrophoresis indicated that GAD is principally multimeric with monomeric forms comprising <3% of the total. Immunoprecipitation and immunoaffinity chromatography experiments were performed with antisera W624 and W883, which were prepared against peptides specific to GAD₆₅ and GAD₆₇, respectively. Immunoprecipitates prepared from cerebellar supernatants with W624 contained both GAD₆₅ and GAD₆₇, whereas some GAD₆₇ was left in the supernatant. In a similar manner, immunoprecipitates prepared with W883 contained both GAD₆₅ and GAD₆₇, whereas some GAD₆₅ remained in the supernatant. In addition, immunoaffinity columns prepared with either W624 or W883 retained both GAD₆₅ and GAD₆₇ even after extensive washing. These results are consistent with the presence of heteromultimers of GAD₆₅ and GAD₆₇ in cerebellum in addition to homomers of each form.     

PIP3-dependent macropinocytosis is incompatible with chemotaxis

In eukaryotic chemotaxis, the mechanisms connecting external signals to the motile apparatus remain unclear. The role of the lipid phosphatidylinositol 3,4,5-trisphosphate (PIP₃) has been particularly controversial. PIP₃ has many cellular roles, notably in growth control and macropinocytosis as well as cell motility. Here we show that PIP₃ is not only unnecessary for Dictyostelium discoideum to migrate toward folate, but actively inhibits chemotaxis. We find that macropinosomes, but not pseudopods, in growing cells are dependent on PIP₃. PIP₃ patches in these cells show no directional bias, and overall only PIP₃-free pseudopods orient up-gradient. The pseudopod driver suppressor of cAR mutations (SCAR)/WASP and verprolin homologue (WAVE) is not recruited to the center of PIP₃ patches, just the edges, where it causes macropinosome formation. Wild-type cells, unlike the widely used axenic mutants, show little macropinocytosis and few large PIP₃ patches, but migrate more efficiently toward folate. Tellingly, folate chemotaxis in axenic cells is rescued by knocking out phosphatidylinositide 3-kinases (PI 3-kinases). Thus PIP₃ promotes macropinocytosis and interferes with pseudopod orientation during chemotaxis of growing cells.     

Morphological indices and otolith microstructure of Atlantic croaker,Micropogonias undulatus,as indicators of habitat quality along an estuarine pollution gradient

Synopsis Atlantic croaker, Micropogonias undulatus, sampled from a transect along a pollution gradient show a trend of declining growth and physical condition. This trend is apparent in the mean size of 0-group croaker, in their recent growth rate measured by marginal otolith increment widths, in longer term growth rate as indicated by relative otolith weights, and in general physical condition as measured by an index of condition of the caudal fin. We suggest that these measures are indicators of stress associated with environmental conditions. Because croaker from different positions along the pollution gradient were distinguishable, it appears that they remain for extended periods within areas of degraded environmental quality.     

Genetic and Structural Analysis of Five Arabidopsis Mutants with Abnormal Root Morphology Generated by the Seed Transformation Method

A number of Arabidopsis thaliana mutants with abnormal rootmorphology have been isolated from a population of Agrobacterium-mediated seed transformants. Five of these mutants identifiedby their abnormal root morphology have been genetically andmorphologically characterised. Two mutants, 5905 and 1767 havedefective cellular elongation, mutant 7203 lacks an endodermalcell layer and mutants 4792 and 7133 exhibit abnormal radialcellular expansion. It is apparent from detailed analysis thatabnormal root morphology is accompanied by abnormal hypocotylphenotype. Genetic analysis has revealed that only one of thesemutants is tagged with a T-DNA insert.Copyright 1994, 1999 AcademicPress Arabidopsis thaliana, root, development, T-DNA insertion mutagenesis, endodermis, elongation