Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 3 de 3
Filter
Add more filters










Database
Language
Publication year range
1.
J Peripher Nerv Syst ; 18(1): 54-8, 2013 Mar.
Article in English | MEDLINE | ID: mdl-23521645

ABSTRACT

In order to develop an efficient, reproducible, and well-tolerated protocol for assessing corneal innervation, 11 normal subjects underwent corneal confocal microscopy (CCM) using a Heidelberg Retinal Tomography III microscope. Five standardized locations were sampled in the left eye and one centrally in the right. The protocol was repeated 1-4 weeks later. A blinded technician measured nerve fiber length (NFL) and tortuosity coefficient (TC). The relationship between image location and NFL and TC was assessed using one-way analysis of variance, and reproducibility determined using relative intertrial variability and intraclass correlation coefficients. NFL reproducibility was maximized by averaging four or more images from the left eye, or one central image from both eyes. TC was less reproducible. CCM is a rapid, well-tolerated, and reproducible method for assessing corneal innervation.


Subject(s)
Cornea/innervation , Microscopy, Confocal , Peripheral Nervous System Diseases/diagnosis , Adult , Female , Humans , Male , Middle Aged , Nerve Fibers/pathology , Peripheral Nervous System Diseases/pathology , Reproducibility of Results
2.
J Cell Biol ; 183(3): 543-54, 2008 Nov 03.
Article in English | MEDLINE | ID: mdl-18955554

ABSTRACT

We have found that MLC-dependent activation of myosin IIB in migrating cells is required to form an extended rear, which coincides with increased directional migration. Activated myosin IIB localizes prominently at the cell rear and produces large, stable actin filament bundles and adhesions, which locally inhibit protrusion and define the morphology of the tail. Myosin IIA forms de novo filaments away from the myosin IIB-enriched center and back to form regions that support protrusion. The positioning and dynamics of myosin IIA and IIB depend on the self-assembly regions in their coiled-coil C terminus. COS7 and B16 melanoma cells lack myosin IIA and IIB, respectively; and show isoform-specific front-back polarity in migrating cells. These studies demonstrate the role of MLC activation and myosin isoforms in creating a cell rear, the segregation of isoforms during filament assembly and their differential effects on adhesion and protrusion, and a key role for the noncontractile region of the isoforms in determining their localization and function.


Subject(s)
Actomyosin/physiology , Cell Movement/physiology , Nonmuscle Myosin Type IIB/physiology , Animals , CHO Cells , COS Cells , Cell Line, Tumor , Chlorocebus aethiops , Cricetinae , Cricetulus , Haplorhini , Humans , Melanoma , Myosin Light Chains/physiology , Nonmuscle Myosin Type IIA/physiology , Nonmuscle Myosin Type IIB/isolation & purification , Phosphorylation
3.
Virology ; 376(1): 42-52, 2008 Jun 20.
Article in English | MEDLINE | ID: mdl-18423509

ABSTRACT

The human cytomegalovirus tegument protein pp71 localizes to the nucleus immediately upon infection, and functions to initiate viral gene expression. Analysis of a series of random insertion mutations revealed that sequences within the mid region (MR) of pp71 are important for localization to the nucleus. Fusion of MR sequences with eGFP revealed that amino acids 94 to 300 were sufficient to target proteins to the nucleus. Random substitution mutagenesis within this domain resulted in two double substitution mutants, pp71P203T/T223M and pp71T228M/L275Q, with a predominantly cytoplasmic localization. Disruption of nuclear targeting resulted in relocalization of the fusion proteins to a distinct perinuclear region. Using tandem mass spectrometry, we determined that threonine 223 can be phosphorylated. Mutation of this residue to a phosphomimetic amino acid resulted in abrogation of nuclear targeting. These results strongly suggest that the intracellular trafficking of pp71 is regulated by phosphorylation.


Subject(s)
Cell Nucleus/chemistry , Cytomegalovirus/physiology , Viral Matrix Proteins/metabolism , Amino Acid Sequence , Amino Acid Substitution/genetics , Artificial Gene Fusion , Genes, Reporter , Green Fluorescent Proteins/genetics , Green Fluorescent Proteins/metabolism , Humans , Molecular Sequence Data , Mutagenesis, Insertional , Mutation, Missense , Nuclear Localization Signals , Phosphorylation , Protein Transport , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/metabolism , Tandem Mass Spectrometry , Threonine/metabolism , Viral Matrix Proteins/chemistry
SELECTION OF CITATIONS
SEARCH DETAIL
...