Association of excitatory amino acid transporters, especially EAAT2, with cholesterol-rich lipid raft microdomains: importance for excitatory amino acid transporter localization and function.
AuthorsButchbach ME, Tian G, Guo H, Lin CL
JournalJ Biol Chem
PubMed ID15187084
In the present study, we investigated the role of membrane cholesterol in the function of glutamate transporters. Depletion of membrane cholesterol by methyl-beta-cyclodextrin resulted in reduced Na(+)-dependent glutamate uptake in primary cortical cultures. Glial glutamate transporter EAAT2-mediated uptake was more sensitive to this effect. Cell surface biotinylation and immunostaining experiments ... More
Identification of a subpopulation of cells with cancer stem cell properties in head and neck squamous cell carcinoma.
AuthorsPrince ME, Sivanandan R, Kaczorowski A, Wolf GT, Kaplan MJ, Dalerba P, Weissman IL, Clarke MF, Ailles LE
JournalProc Natl Acad Sci U S A
PubMed ID17210912
'Like many epithelial tumors, head and neck squamous cell carcinoma (HNSCC) contains a heterogeneous population of cancer cells. We developed an immunodeficient mouse model to test the tumorigenic potential of different populations of cancer cells derived from primary, unmanipulated human HNSCC samples. We show that a minority population of CD44(+) ... More
Prototype protein assembly as scaffold for time-resolved fluoroimmuno assays.
AuthorsBarnhill HN, Claudel-Gillet S, Ziessel R, Charbonnière LJ, Wang Q
JournalJ Am Chem Soc
PubMed ID17542581
'Turnip yellow mosaic virus (TYMV) is an icosahedral plant virus with an average diameter of 28 nm and can be isolated in gram quantities from turnip or Chinese cabbage inexpensively. In this study, it was selected as a prototype bionanoparticle for time-resolved fluoroimmuno assay (TRFIA). Two types of reactive amino ... More
Protein cargo delivery properties of cell-penetrating peptides. A comparative study.
AuthorsSäälik P, Elmquist A, Hansen M, Padari K, Saar K, Viht K, Langel U, Pooga M
JournalBioconjug Chem
PubMed ID15546190
'Application of cell-penetrating peptides for delivering various hydrophilic macromolecules with biological function into cells has gained much attention in recent years. We compared the protein transduction efficiency of four cell-penetrating peptides: penetratin, Tat peptide, transportan, and pVEC and studied the effects of various medium parameters on the uptake. Depletion of ... More
The role of alpha and beta chains in ligand recognition by beta 7 integrins.
AuthorsHiggins JM, Cernadas M, Tan K, Irie A, Wang J, Takada Y, Brenner MB
JournalJ Biol Chem
PubMed ID10837471
'Integrins alpha(E)beta(7) and alpha(4)beta(7) are involved in localization of leukocytes at mucosal sites. Although both alpha(E)beta(7) and alpha(4)beta(7) utilize the beta(7) chain, they have distinct binding specificities for E-cadherin and mucosal addressin cell adhesion molecule-1 (MAdCAM-1), respectively. We found that mutation of the metal ion-dependent adhesion site (MIDAS) in the ... More
Rapid degeneration of noncoding DNA regions surrounding SlAP3X/Y after recombination suppression in the dioecious plant Silene latifolia.
'Silene latifolia is a dioecious plant with heteromorphic XY sex chromosomes. Previous studies of sex chromosome-linked genes have suggested a gradual divergence between the X-linked and the Y-linked genes in proportion to the distance from the pseudoautosomal region. However, such a comparison has yet to be made for the noncoding ... More
Mast cell-deficient Kit(W-sh)
AuthorsMichel A, Schüler A, Friedrich P, Döner F, Bopp T, Radsak M, Hoffmann M, Relle M, Distler U, Kuharev J, Tenzer S, Feyerabend TB, Rodewald HR, Schild H, Schmitt E, Becker M, Stassen M,
JournalJ Immunol
PubMed ID23636054
'Mast cell-deficient Kit(W-sh) '
Axons giving rise to the palisade endings of feline extraocular muscles display motor features.
AuthorsZimmermann L, Morado-Díaz CJ, Davis-López de Carrizosa MA, de la Cruz RR, May PJ, Streicher J, Pastor ÁM, Blumer R,
JournalJ Neurosci
PubMed ID23407938
'Palisade endings are nerve specializations found in the extraocular muscles (EOMs) of mammals, including primates. They have long been postulated to be proprioceptors. It was recently demonstrated that palisade endings are cholinergic and that in monkeys they originate from the EOM motor nuclei. Nevertheless, there is considerable difference of opinion ... More
Non-covalent delivery of proteins into mammalian cells.
AuthorsLoudet A, Han J, Barhoumi R, Pellois JP, Burghardt RC, Burgess K,
JournalOrg Biomol Chem
PubMed ID19039359
Substances that mediate the import of proteins into cells,
In vivo models for studying mast cell-dependent responses to bacterial infection.
AuthorsShelburne CP, McLachlan JB, Abraham SN,
JournalMethods Mol Biol
PubMed ID16110170
Mast cells are a critical component of host defense against bacterial infections. Activation of these cells during infection induces both innate and adaptive aspects of protective immunity needed for the elimination of the bacteria and survival of the host. These functional roles for the mast cell have been principally characterized ... More
Discovery of aminoacyl-tRNA synthetase activity through cell-surface display of noncanonical amino acids.
The incorporation of noncanonical amino acids into recombinant proteins in Escherichia coli can be facilitated by the introduction of new aminoacyl-tRNA synthetase activity into the expression host. We describe here a screening procedure for the identification of new aminoacyl-tRNA synthetase activity based on the cell surface display of noncanonical amino ... More
VEGF gene delivery to muscle: potential role for vasculogenesis in adults.
AuthorsSpringer ML, Chen AS, Kraft PE, Bednarski M, Blau HM
JournalMol Cell
PubMed ID9844628
Constitutive expression of VEGF after implantation of genetically engineered myoblasts into non-ischemic muscle led to an increase in vascular structures. Previously, effects of VEGF delivery to adult muscle have only been reported in ischemic tissues. The resulting vascular structures were reminiscent of those formed during embryonic vasculogenesis, rather than angiogenesis, ... More
Aging-associated shifts in functional status of mast cells located by adult and aged mesenteric lymphatic vessels.
AuthorsChatterjee V, Gashev AA,
JournalAm J Physiol Heart Circ Physiol
PubMed ID22796537
We had previously proposed the presence of permanent stimulatory influences in the tissue microenvironment surrounding the aged mesenteric lymphatic vessels (MLV), which influence aged lymphatic function. In this study, we performed immunohistochemical labeling of proteins known to be present in mast cells (mast cell tryptase, c-kit, prostaglandin D(2) synthase, histidine ... More
Polyethyleneimine as a transmembrane carrier of fluorescently labeled proteins and antibodies.
AuthorsDidenko VV, Ngo H, Baskin DS,
JournalAnal Biochem
PubMed ID16095551
Polyethyleneimine (PEI) has been used previously as a nonviral DNA transfer vector. In this article, we demonstrate its use as a vehicle for transmembrane delivery of proteins in cell culture conditions. Linking proteins to PEI required no other treatment beyond mixing them with PEI. The bond between PEI and protein ... More
Advanced glycation end products induce a prothrombotic phenotype in mice via interaction with platelet CD36.
AuthorsZhu W, Li W, Silverstein RL,
JournalBlood
PubMed ID22431576
Diabetes mellitus has been associated with platelet hyperreactivity, which plays a central role in the hyperglycemia-related prothrombotic phenotype. The mechanisms responsible for this phenomenon are not established. In the present study, we investigated the role of CD36, a class-B scavenger receptor, in this process. Using both in vitro and in ... More
Phenotypic characterization of gastric sensory neurons in mice.
AuthorsBielefeldt K, Zhong F, Koerber HR, Davis BM,
JournalAm J Physiol Gastrointest Liver Physiol
PubMed ID16728726
Recent studies suggest that the capsaicin receptor [transient receptor potential vanilloid (TRPV)1] may play a role in visceral mechanosensation. To address the potential role of TRPV1 in vagal sensory neurons, we developed a new in vitro technique allowing us to determine TRPV1 expression directly in physiologically characterized gastric sensory neurons. ... More
Heat sensitization in skin and muscle nociceptors expressing distinct combinations of TRPV1 and TRPV2 protein.
AuthorsRau KK, Jiang N, Johnson RD, Cooper BY,
JournalJ Neurophysiol
PubMed ID17287441
Recordings were made from small and medium diameter dorsal root ganglia (DRG) neurons that expressed transient receptor potential (TRP) proteins. Physiologically characterized skin nociceptors expressed either TRPV1 (type 2) or TRPV2 (type 4) in isolation. Other nociceptors co-expressed both TRP proteins and innervated deep tissue sites (gastrocnemius muscle, distal colon; ... More
Direct fluorescent staining and analysis of proteins on microspheres using CBQCA.
AuthorsGraves SW, Woods TA, Kim H, Nolan JP
JournalCytometry A
PubMed ID15812850
BACKGROUND: General methods for accurate determination of microsphere surface protein loading are needed for applications from protein arrays to molecular assembly studies. Current methods include bulk absorption measurements of stained microspheres or use of known fluorescently tagged binding partners, which limit sensitivity and general applicability, respectively. METHODS: Microspheres bearing covalently ... More
Double-label confocal laser-scanning microscopy, image restoration, and real-time three-dimensional reconstruction to study axons in the central nervous system and their contacts with target neurons.
AuthorsWouterlood FG, van Haeften T, Blijleven N, Pérez-Templado P, Pérez-Templado H
JournalAppl Immunohistochem Mol Morphol
PubMed ID11893043
The current double tracing-double confocal laser-scanning method was developed to reconstruct identified nerve fibers and their contacts with identified target neurons in the rat brain in three dimensions. It intends to fill the gap between conventional light microscopic and electron microscopic neuroanatomic tracing. The steps involved are as follows: (1) ... More
An endoplasmic reticulum stress-specific caspase cascade in apoptosis. Cytochrome c-independent activation of caspase-9 by caspase-12.
Activation of caspase-12 from procaspase-12 is specifically induced by insult to the endoplasmic reticulum (ER) (Nakagawa, T., Zhu, H., Morishima, N., Li, E., Xu, J., Yankner, B. A., and Yuan, J. (2000) Nature 403, 98-103), yet the functional consequences of caspase-12 activation have been unclear. We have shown that recombinant ... More