Cast: a novel protein of the cytomatrix at the active zone of synapses that forms a ternary complex with RIM1 and munc13-1

The cytomatrix at the active zone (CAZ) has been implicated in defining the site of Ca2+-dependent exocytosis of neurotransmitter. We have identified here a novel CAZ protein of ∼120 kD from rat brain and named it CAST (CAZ-associated structural protein). CAST had no transmembrane segment, but had four coiled-coil domains and a putative COOH-terminal consensus motif for binding to PDZ domains. CAST was localized at the CAZ of conventional synapses of mouse brain. CAST bound directly RIM1 and indirectly Munc13-1, presumably through RIM1, forming a ternary complex. RIM1 and Munc13-1 are CAZ proteins implicated in Ca2+-dependent exocytosis of neurotansmitters. Bassoon, another CAZ protein, was also associated with this ternary complex. These results suggest that a network of protein–protein interactions among the CAZ proteins exists at the CAZ. At the early stages of synapse formation, CAST was expressed and partly colocalized with bassoon in the axon shaft and the growth cone. The vesicles immunoisolated by antibassoon antibody–coupled beads contained not only bassoon but also CAST and RIM1. These results suggest that these CAZ proteins are at least partly transported on the same vesicles during synapse formation

Harbour Grace Standard

The Harbor Grace Standard was published from December 1859 to 1936[?], providing coverage of the Conception Bay area as well as other news. The frequency was weekly save for a brief semiweekly interlude between 1888-94 and there is a publication gap between 1868 and February 1871. Many issues are missing in the later years.Alternative title from common misspelling, as the city's modern name features a "u.

Tumour-initiating ability of HSAEC_4T53RD cells.

<p>Tumour-initiating ability of HSAEC_4T53RD cells.</p

Expression of differentiation markers in both <i>in vivo</i> xenografts and <i>in vitro</i> differentiation cultures.

<p>Sections of xenografts formed from a single-cell-derived HSAEC_4T53RD clone were subjected to staining for (A) K5 (blue), K8 (green) and p63 (red); and (B) SCGB1A1 (green) and MUC5AC (red). Spheres formed by one of the single-cell-derived HSAEC_4T53RD clones were subjected to staining for (C) K5 (blue), K8 (green) and p63 (red); and (D) SCGB1A1 (green) and MUC5AC (red).</p

Phenotype of HSAEC_4T53RD xenografts.

<p>Formalin-fixed, paraffin-embedded sections (4-μm thick) of xenografts derived from HSAEC_4T53RD cells were subjected to staining with (A) haematoxylin and eosin or (B) alcian blue; or with antibodies against (C) cytokeratins (AE1/AE3), (D) p63, (E) alpha-SMA (blue) and p53 (brown). (F) HSAEC_4T53RD cells cultured <i>in vitro</i> on collagen-coated dishes in serum-free SAGM medium were stained with anti-p63 antibody (green) and phalloidin (red).</p