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Charles Watson

Publications and source records attributed to Charles Watson.

9 recordsLinked to original sources
PubMed2006

A stereotaxic MRI template set for the rat brain with tissue class distribution maps and co-registered anatomical atlas: application to pharmacological MRI.

We describe a stereotaxic rat brain MRI template set with a co-registered digital anatomical atlas and illustrate its application to the analysis of a pharmacological MRI (phMRI) study of apomorphine. The template set includes anatomical images and tissue class probability maps for brain parenchyma and cerebrospinal fluid (CSF). These facilitate the use of standard fMRI software for spatial normalisation and tissue segmentation of rat brain data. A volumetric reconstruction of the Paxinos and Watson rat brain atlas is also co-localised with the template, enabling the atlas structure and stereotaxic coordinates corresponding to a feature within a statistical map to be interactively reported, facilitating the localisation of functional effects. Moreover, voxels falling within selected brain structures can be combined to define anatomically based 3D volumes of interest (VOIs), free of operator bias. As many atlas structures are small relative to the typical resolution of phMRI studies, a mechanism for defining composite structures as agglomerations of individual atlas structures is also described. This provides a simple and robust means of interrogating structures that are otherwise difficult to delineate and an objective framework for comparing and classifying compounds based on an anatomical profile of their activity. These developments allow a closer alignment of pre-clinical and clinical analysis techniques.

Animals↗
PubMed2006

Proteomic identification and functional characterization of a novel ARF6 GTPase-activating protein, ACAP4.

ARF6 GTPase is a conserved regulator of membrane trafficking and actin-based cytoskeleton dynamics at the leading edge of migrating cells. A key determinant of ARF6 function is the lifetime of the GTP-bound active state, which is orchestrated by GTPase-activating protein (GAP) and GTP-GDP exchanging factor. However, very little is known about the molecular mechanisms underlying ARF6-mediated cell migration. To systematically analyze proteins that regulate ARF6 activity during cell migration, we performed a proteomic analysis of proteins selectively bound to active ARF6 using mass spectrometry and identified a novel ARF6-specific GAP, ACAP4. ACAP4 encodes 903 amino acids and contains two coiled coils, one pleckstrin homology domain, one GAP motif, and two ankyrin repeats. Our biochemical characterization demonstrated that ACAP4 has a phosphatidylinositol 4,5-bisphosphate-dependent GAP activity specific for ARF6. The co-localization of ACAP4 with ARF6 occurred in ruffling membranes formed upon AIF(4) and epidermal growth factor stimulation. ACAP4 overexpression limited the recruitment of ARF6 to the membrane ruffles in the absence of epidermal growth factor stimulation. Expression of GTP hydrolysis-resistant ARF6(Q67L) resulted in accumulations of ACAP4 and ARF6 in the cytoplasmic membrane, suggesting that GTP hydrolysis is required for the ARF6-dependent membrane remodeling. Significantly the depletion of ACAP4 by small interfering RNA or inhibition of ARF6 GTP hydrolysis by overexpressing GAP-deficient ACAP4 suppressed ARF6-dependent cell migration in wound healing, demonstrating the importance of ACAP4 in cell migration. Thus, our study sheds new light on the biological function of ARF6-mediated cell migration.

ADP-Ribosylation Factor 6↗
PubMed2006

Neuroanatomical affiliation visualization-interface system.

A number of knowledge management systems have been developed to allow users to have access to large quantity of neuroanatomical data. The advent of three-dimensional (3D) visualization techniques allows users to interact with complex 3D object. In order to better understand the structural and functional organization of the brain, we present Neuroanatomical Affiliations Visualization-Interface System (NAVIS) as the original software to see brain structures and neuroanatomical affiliations in 3D. This version of NAVIS has made use of the fifth edition of "The Rat Brain in Stereotaxic coordinates" (Paxinos and Watson, 2005). The NAVIS development environment was based on the scripting language name Python, using visualization toolkit (VTK) as 3D-library and wxPython for the graphic user interface. The following manuscript is focused on the nucleus of the solitary tract (Sol) and the set of affiliated structures in the brain to illustrate the functionality of NAVIS. The nucleus of the Sol is the primary relay center of visceral and taste information, and consists of 14 distinct subnuclei that differ in cytoarchitecture, chemoarchitecture, connections, and function. In the present study, neuroanatomical projection data of the rat Sol were collected from selected literature in PubMed since 1975. Forty-nine identified projection data of Sol were inserted in NAVIS. The standard XML format used as an input for affiliation data allows NAVIS to update data online and/or allows users to manually change or update affiliation data. NAVIS can be extended to nuclei other than Sol.

Animals↗
PubMed2003

Characterization of protein kinase A-mediated phosphorylation of ezrin in gastric parietal cell activation.

Gastric ezrin was initially identified as a phosphoprotein associated with parietal cell activation. To explore the nature of ezrin phosphorylation, proteins from resting and secreting gastric glands were subjected to two-dimensional SDS-PAGE. Histamine triggers acid secretion and a series of acidic isoforms of ezrin on two-dimensional SDS-PAGE. Mass spectrometric analysis of these acidic ezrin spots induced by stimulation suggests that Ser66 is phosphorylated. To determine whether Ser66 is a substrate of protein kinase A (PKA), recombinant proteins of ezrin, both wild type and S66A mutant, were incubated with the catalytic subunit of PKA and [32P]ATP. Incorporation of 32P into wild type but not the mutant ezrin verified that Ser66 is a substrate of PKA. In addition, expression of S66A mutant ezrin in cultured parietal cells attenuates the dilation of apical vacuolar membrane associated with stimulation by histamine, indicating that PKA-mediated phosphorylation of ezrin is necessary for acid secretion. In fact, expression of phosphorylation-like S66D mutant in parietal cells mimics histamine-stimulated apical vacuole remodeling. Further examination of H,K-ATPase distribution revealed a blockade of stimulation-induced proton pump mobilization in S66A but not S66D ezrin-expressing parietal cells. These data suggest that PKA-mediated phosphorylation of ezrin plays an important role in mediating the remodeling of the apical membrane cytoskeleton associated with acid secretion in parietal cells.

Adenosine Triphosphate↗
PubMed2003

Myosin II is present in gastric parietal cells and required for lamellipodial dynamics associated with cell activation.

Nonmuscle myosin II has been shown to participate in organizing the actin cytoskeleton in polarized epithelial cells. Vectorial acid secretion in cultured parietal cells involves translocation of proton pumps from cytoplasmic vesicular membranes to the apical plasma membrane vacuole with coordinated lamellipodial dynamics at the basolateral membrane. Here we identify nonmuscle myosin II in rabbit gastric parietal cells. Western blots with isoform-specific antibodies indicate that myosin IIA is present in both cytosolic and particulate membrane fractions whereas the IIB isoform is associated only with particulate fractions. Immunofluorescent staining demonstrates that myosin IIA is diffusely located throughout the cytoplasm of resting parietal cells. However, after stimulation, myosin IIA is rapidly redistributed to lamellipodial extensions at the cell periphery; virtually all the cytoplasmic myosin IIA joins the newly formed basolateral membrane extensions. 2,3-Butanedione monoximine (BDM), a myosin-ATPase inhibitor, greatly diminishes the lamellipodial dynamics elicited by stimulation and retains the pattern of myosin IIA cytoplasmic staining. However, BDM had no apparent effect on the stimulation associated redistribution of H,K-ATPase from a cytoplasmic membrane compartment to apical membrane vacuoles. The myosin light chain kinase inhibitor 1-(5-iodonaphthalene-1-sulfonyl)-1H-hexahydro-1,4-diazepine (ML-7) also did not alter the stimulation-associated recruitment of H,K-ATPase to apical membrane vacuoles, but unlike BDM it had relatively minor inhibitory effects on lamellipodial dynamics. We conclude that specific disruption of the basolateral actomyosin cytoskeleton has no demonstrable effect on recruitment of H,K-ATPase-rich vesicles into the apical secretory membrane. However, myosin II plays an important role in regulating lamellipodial dynamics and cortical actomyosin associated with parietal cell activation.

Aminopyrine↗
PubMed2003

Polarized distribution of IQGAP proteins in gastric parietal cells and their roles in regulated epithelial cell secretion.

Actin cytoskeleton plays an important role in the establishment of epithelial cell polarity. Cdc42, a member of Rho GTPase family, modulates actin dynamics via its regulators, such as IQGAP proteins. Gastric parietal cells are polarized epithelial cells in which regulated acid secretion occurs in the apical membrane upon stimulation. We have previously shown that actin isoforms are polarized to different membrane domains and that the integrity of the actin cytoskeleton is essential for acid secretion. Herein, we show that Cdc42 is preferentially distributed to the apical membrane of gastric parietal cells. In addition, we revealed that two Cdc42 regulators, IQGAP1 and IQGAP2, are present in gastric parietal cells. Interestingly, IQGAP2 is polarized to the apical membrane of the parietal cells, whereas IQGAP1 is mainly distributed to the basolateral membrane. An IQGAP peptide that competes with full-length IQGAP proteins for Cdc42-binding in vitro also inhibits acid secretion in streptolysin-O-permeabilized gastric glands. Furthermore, this peptide disrupts the association of IQGAP and Cdc42 with the apical actin cytoskeleton and prevents the apical membrane remodeling upon stimulation. We propose that IQGAP2 forms a link that associates Cdc42 with the apical cytoskeleton and thus allows for activation of polarized secretion in gastric parietal cells.

Animals↗
PubMed2002

Australia declared polio free.

For Australia to be declared polio free, evidence of the absence of circulation of wild poliovirus was required by the Regional Commission for the Certification of Eradication of Poliomyelitis in the Western Pacific in August 2000. Data on surveillance of poliomyelitis, acute flaccid paralysis (AFP), vaccine associated paralytic polio and enteroviruses were provided to document the absence of circulation of wild poliovirus. The last wild poliomyelitis virus case in Australia was in 1972. AFP surveillance has improved since it was initiated in 1995 and achieved a rate of 0.94 per 100,000 population in 1999. No wild polioviruses have been isolated from stool samples of AFP cases. Australia has in place a comprehensive network of laboratories for enterovirus surveillance and this provides further evidence for the absence of wild poliovirus infection. The immunisation coverage in the country has been over 80 per cent over the last 3 years. If there were an importation of a case of poliomyelitis into Australia, a national outbreak response would be coordinated through the Communicable Diseases Network Australia. Plans for containment of laboratory stocks of wild poliovirus are being implemented. The evidence provided was sufficient to satisfy the Regional Commission that there was no wild poliovirus circulating in the region and enabled Australia to be declared polio free on October 29, 2000 along with the other 36 countries in the Western Pacific Region. Australia must remain vigilant against importations of wild poliovirus from endemic countries and maintain high immunisation coverage and sensitive surveillance systems.

Adolescent↗