The CRE/CREB pathway is transiently expressed in thalamic circuit development and contributes to refinement of retinogeniculate axons.
AuthorsPham TA, Rubenstein JL, Silva AJ, Storm DR, Stryker MP
JournalNeuron
PubMed ID11516398
'The development of precise connections in the mammalian brain proceeds through refinement of initially diffuse patterns, a process that occurs largely within critical developmental windows. To elucidate the molecular pathways that orchestrate these early periods of circuit remodeling, we have examined the role of a calcium- and cAMP-regulated transcriptional pathway. ... More
Development of a delivery vehicle for intracellular transport of botulinum neurotoxin antagonists.
AuthorsGoodnough MC, Oyler G, Fishman PS, Johnson EA, Neale EA, Keller JE, Tepp WH, Clark M, Hartz S, Adler M
JournalFEBS Lett
PubMed ID11904143
'A targeted delivery vehicle (DV) was developed for intracellular transport of emerging botulinum neurotoxin (BoNT) antagonists. The DV consisted of the isolated heavy chain (HC) of BoNT/A coupled to a 10-kDa amino dextran via the heterobifunctional linker 3-(2-pyridylthio)-propionyl hydrazide. The HC served to target BoNT-sensitive cells and promote internalization of ... More
Loss of the chloride channel ClC-7 leads to lysosomal storage disease and neurodegeneration.
AuthorsKasper D, Planells-Cases R, Fuhrmann JC, Scheel O, Zeitz O, Ruether K, Schmitt A, Poët M, Steinfeld R, Schweizer M, Kornak U, Jentsch TJ
JournalEMBO J
PubMed ID15706348
'ClC-7 is a chloride channel of late endosomes and lysosomes. In osteoclasts, it may cooperate with H(+)-ATPases in acidifying the resorption lacuna. In mice and man, loss of ClC-7 or the H(+)-ATPase a3 subunit causes osteopetrosis, a disease characterized by defective bone resorption. We show that ClC-7 knockout mice additionally ... More
Dermal microvascular endothelial cells express the 180-kDa macrophage mannose receptor in situ and in vitro.
AuthorsGröger M, Holnthoner W, Maurer D, Lechleitner S, Wolff K, Mayr BB, Lubitz W, Petzelbauer P
JournalJ Immunol
PubMed ID11067894
'Expression of the 180-kDa mannose receptor (MR) is mainly found on cells of the macrophage lineage. MR mediates the uptake of micro-organisms and host-derived glycoproteins. We demonstrate that endothelium of the human skin in situ and dermal microvascular endothelial cells (DMEC) in vitro expressed MR at both the protein and ... More
Characterization of cellular optoporation with distance.
'We have developed and characterized cellular optoporation with visible wavelengths of light using standard uncoated glass cover slips as the absorptive media. A frequency-doubled Nd:YAG laser pulse was focused at the interface of the glass surface and aqueous buffer, creating a stress wave and transiently permeabilizing nearby cells. Following optoporation ... More
The recycling endosome of Madin-Darby canine kidney cells is a mildly acidic compartment rich in raft components.
AuthorsGagescu R, Demaurex N, Parton RG, Hunziker W, Huber LA, Gruenberg J
JournalMol Biol Cell
PubMed ID10930469
'We present a biochemical and morphological characterization of recycling endosomes containing the transferrin receptor in the epithelial Madin-Darby canine kidney cell line. We find that recycling endosomes are enriched in molecules known to regulate transferrin recycling but lack proteins involved in early endosome membrane dynamics, indicating that recycling endosomes are ... More
Calpain activity promotes the sealing of severed giant axons.
'A barrier (seal) must form at the cut ends of a severed axon if a neuron is to survive and eventually regenerate. Following severance of crayfish medial giant axons in physiological saline, vesicles accumulate at the cut end and form a barrier (seal) to ion and dye diffusion. In contrast, ... More
The stress-induced MAP kinase p38 regulates endocytic trafficking via the GDI:Rab5 complex.
AuthorsCavalli V, Vilbois F, Corti M, Marcote MJ, Tamura K, Karin M, Arkinstall S, Gruenberg J
JournalMol Cell
PubMed ID11239470
Early endocytic membrane traffic is regulated by the small GTPase Rab5, which cycles between GTP- and GDP-bound states as well as between membrane and cytosol. The latter cycle depends on GDI, which functions as a Rab vehicle in the aqueous environment of the cytosol. Here, we report that formation of ... More
Monitoring autophagy in lysosomal storage disorders.
AuthorsRaben N, Shea L, Hill V, Plotz P,
JournalMethods Enzymol
PubMed ID19216919
Lysosomes are the final destination of the autophagic pathway. It is in the acidic milieu of the lysosomes that autophagic cargo is metabolized and recycled. One would expect that diseases with primary lysosomal defects would be among the first systems in which autophagy would be studied. In reality, this is ... More
Lysosomal storage disease upon disruption of the neuronal chloride transport protein ClC-6.
AuthorsPoët M, Kornak U, Schweizer M, Zdebik AA, Scheel O, Hoelter S, Wurst W, Schmitt A, Fuhrmann JC, Planells-Cases R, Mole SE, Hübner CA, Jentsch TJ
JournalProc Natl Acad Sci U S A
PubMed ID16950870
Mammalian CLC proteins function as Cl(-) channels or as electrogenic Cl(-)/H(+) exchangers and are present in the plasma membrane and intracellular vesicles. We now show that the ClC-6 protein is almost exclusively expressed in neurons of the central and peripheral nervous systems, with a particularly high expression in dorsal root ... More
Membrane perforations inhibit lysosome fusion by altering pH and calcium in Listeria monocytogenes vacuoles.
AuthorsShaughnessy LM, Hoppe AD, Christensen KA, Swanson JA
JournalCell Microbiol
PubMed ID16611227
Listeria monocytogenes (Lm) evade microbicidal defences inside macrophages by secreting a pore-forming cytolysin listeriolysin O (LLO), which allows Lm to escape vacuoles. LLO also inhibits Lm vacuole fusion with lysosomes, which indicates LLO alters vacuole chemistry prior to release of Lm into cytoplasm. Using fluorescent probes to measure membrane permeability, ... More
Transmitter-evoked local calcium release stabilizes developing dendrites.
AuthorsLohmann C, Myhr KL, Wong RO
JournalNature
PubMed ID12110889
In the central nervous system, dendritic arborizations of neurons undergo dynamic structural remodelling during development. Processes are elaborated, maintained or eliminated to attain the adult pattern of synaptic connections. Although neuronal activity influences this remodelling, it is not known how activity exerts its effects. Here we show that neurotransmission-evoked calcium ... More
An inhibitory role of Rho in the vasopressin-mediated translocation of aquaporin-2 into cell membranes of renal principal cells.
AuthorsKlussmann E, Tamma G, Lorenz D, Wiesner B, Maric K, Hofmann F, Aktories K, Valenti G, Rosenthal W
JournalJ Biol Chem
PubMed ID11278652
Vasopressin regulates water reabsorption in renal collecting duct principal cells by a cAMP-dependent translocation of the water channel aquaporin-2 (AQP2) from intracellular vesicles into the cell membrane. In the present work primary cultured inner medullary collecting duct cells were used to study the role of the proteins of the Rho ... More
The invagination of excess surface area by shrinking neurons.
AuthorsMorris CE, Wang JA, Markin VS
JournalBiophys J
PubMed ID12829478
Over most of their surface, neurons are surrounded by a narrow extracellular gap across which they make adhesive cell-cell contacts. Thus constrained, how do they regulate their geometry when osmotically perturbed? Specifically, are there any interesting consequences of local osmosis in such conditions? Using confocal imaging of shrinking neurons in ... More
A real-time ratiometric method for the determination of molecular oxygen inside living cells using sol-gel-based spherical optical nanosensors with applications to rat C6 glioma.
AuthorsXu H, Aylott JW, Kopelman R, Miller TJ, Philbert MA
JournalAnal Chem
PubMed ID11569801
The first sol-gel-based, ratiometric, optical nanosensors, or sol-gel probes encapsulated by biologically localized embedding (PEBBLEs), are made and demonstrated here to enable reliable, real-time measurements of subcellular molecular oxygen. Sensors were made using a modified Stöber method, with poly(ethylene glycol) as a steric stabilizer. The radii of these spherical PEBBLE ... More
Crayfish medial giant axons (MGAs) transected in physiological saline form vesicles which interact with each other, pre-existing vesicles, and/or with the plasmalemma to form an electrical and a physical barrier that seals a cut axonal end within 60 min. The formation of this barrier (seal) was assessed by measuring the ... More
Ryanodine receptor type I and nicotinic acid adenine dinucleotide phosphate receptors mediate Ca2+ release from insulin-containing vesicles in living pancreatic beta-cells (MIN6).
AuthorsMitchell KJ, Lai FA, Rutter GA
JournalJ Biol Chem
PubMed ID12538591
We have demonstrated recently (Mitchell, K. J., Pinton, P., Varadi, A., Tacchetti, C., Ainscow, E. K., Pozzan, T., Rizzuto, R., and Rutter, G. A. (2001) J. Cell Biol. 155, 41-51) that ryanodine receptors (RyR) are present on insulin-containing secretory vesicles. Here we show that pancreatic islets and derived beta-cell lines ... More
Glass needle-mediated microinjection of macromolecules and transgenes into primary human mesenchymal stem cells.
AuthorsTsulaia TV, Prokopishyn NL, Yao A, Carsrud ND, Carou MC, Brown DB, Davis BR, Yannariello-Brown J
JournalJ Biomed Sci
PubMed ID12711860
Human mesenchymal stem cells (hMSCs) are multipotent cells that can differentiate into various tissue types, including bone, cartilage, tendon, adipocytes, and marrow stroma, making them potentially useful for human cell and gene therapies. Our objective was to demonstrate the utility of glass needle-mediated microinjection as a method to deliver macromolecules ... More
Spinning disk confocal microscopy of live, intraerythrocytic malarial parasites. 2. Altered vacuolar volume regulation in drug resistant malaria.
In the previous paper [Gligorijevic, B., et al. (2006) Biochemistry 45, pp 12400-12410], we reported on a customized Nipkow spinning disk confocal microscopy (SDCM) system and its initial application to DIC imaging of hemozoin within live, synchronized, intraerythrocytic Plasmodium falciparum malarial parasites. In this paper, we probe the biogenesis as ... More
Role of COPI in phagosome maturation.
AuthorsBotelho RJ, Hackam DJ, Schreiber AD, Grinstein S
JournalJ Biol Chem
PubMed ID10748188
Phagosomes mature by sequentially fusing with endosomes and lysosomes. Vesicle budding is presumed to occur concomitantly, mediating the retrieval of plasmalemmal components and the regulation of phagosomal size. We analyzed whether fission of vesicles from phagosomes requires COPI, a multimeric complex known to be involved in budding from the Golgi ... More
Post-Golgi carriers, not lysosomes, confer lysosomal properties to pre-degradative organelles in normal and dystrophic axons.
Authors
JournalCell Rep
PubMed ID33910020
mTOR regulates lysosomal ATP-sensitive two-pore Na(+) channels to adapt to metabolic state.
Authors
JournalCell
PubMed ID23394946
Impaired lysosomal acidification triggers iron deficiency and inflammation in vivo.
Authors
JournalElife
PubMed ID31793879
A method to experimentally clamp leaf water content to defined values to assess its effects on apoplastic pH.
Authors
JournalPlant Methods
PubMed ID35644610
A family of PIKFYVE inhibitors with therapeutic potential against autophagy-dependent cancer cells disrupt multiple events in lysosome homeostasis.