Sharon F. Suchy, Robert L. Nussbaum 

Slides:



Advertisements
Similar presentations
Ezrin Is Down-Regulated in Diabetic Kidney Glomeruli and Regulates Actin Reorganization and Glucose Uptake via GLUT1 in Cultured Podocytes  Anita A. Wasik,
Advertisements

Volume 15, Issue 3, Pages (March 2007)
Volume 70, Issue 7, Pages (October 2006)
Figure 8. Colocalization of β-Arrestin2-GFP with Internalized wt and S(483–498)A Mutant PTHRs HEK-wt PTHR and HEK-S(483–498)A PTHR cells were transiently.
Okadaic-Acid-Induced Inhibition of Protein Phosphatase 2A Produces Activation of Mitogen-Activated Protein Kinases ERK1/2, MEK1/2, and p70 S6, Similar.
Volume 128, Issue 1, Pages (January 2005)
Dongfang Liu, Mary E. Peterson, Eric O. Long  Immunity 
Pericentrosomal Localization of the TIG3 Tumor Suppressor Requires an N-Terminal Hydrophilic Region Motif  Tiffany M. Scharadin, Gautam Adhikary, Kristin.
Visualization of ER-to-Golgi Transport in Living Cells Reveals a Sequential Mode of Action for COPII and COPI  Suzie J. Scales, Rainer Pepperkok, Thomas.
The renal Na+/phosphate cotransporter NaPi-IIa is internalized via the receptor- mediated endocytic route in response to parathyroid hormone  D. Bacic,
Fibroblast Activation Protein: Differential Expression and Serine Protease Activity in Reactive Stromal Fibroblasts of Melanocytic Skin Tumors  Margit.
A Nonsense Mutation in the Human Homolog of Drosophila rogdi Causes Kohlschutter– Tonz Syndrome  Adi Mory, Efrat Dagan, Barbara Illi, Philippe Duquesnoy,
Linda Vi, Stellar Boo, Samar Sayedyahossein, Randeep K
Actomyosin Tension Exerted on the Nucleus through Nesprin-1 Connections Influences Endothelial Cell Adhesion, Migration, and Cyclic Strain-Induced Reorientation 
Volume 70, Issue 7, Pages (October 2006)
Volume 126, Issue 7, Pages (June 2004)
Requirement of VPS33B, a member of the Sec1/Munc18 protein family, in megakaryocyte and platelet α-granule biogenesis by Bryan Lo, Ling Li, Paul Gissen,
Palladin is a dynamic actin-associated protein in podocytes
Mutations in SLC33A1 Cause a Lethal Autosomal-Recessive Disorder with Congenital Cataracts, Hearing Loss, and Low Serum Copper and Ceruloplasmin  Peter.
Carlos Rio, Heather I Rieff, Peimin Qi, Gabriel Corfas  Neuron 
Dongfang Liu, Mary E. Peterson, Eric O. Long  Immunity 
Volume 68, Issue 6, Pages (December 2005)
Plectin-Isoform-Specific Rescue of Hemidesmosomal Defects in Plectin (–/–) Keratinocytes  Kerstin Andrä, Iris Kornacker, Almut Jörgl, Michael Zörer, Daniel.
Volume 92, Issue 8, Pages (April 2007)
Pdlim2 is a novel actin-regulating protein of podocyte foot processes
Volume 11, Issue 18, Pages (September 2001)
Emma L. Baple, Reza Maroofian, Barry A. Chioza, Maryam Izadi, Harold E
Ann-Marie Broome, Richard L. Eckert, PhD 
Localization of Calcineurin/NFAT in Human Skin and Psoriasis and Inhibition of Calcineurin/NFAT Activation in Human Keratinocytes by Cyclosporin A  Wael.
Kinesin and Kinectin Can Associate with the Melanosomal Surface and Form a Link with Microtubules in Normal Human Melanocytes1  Garnet Vancoillie, Jo.
Colocalization of Kindlin-1, Kindlin-2, and Migfilin at Keratinocyte Focal Adhesion and Relevance to the Pathophysiology of Kindler Syndrome  J.E. Lai-Cheong,
Volume 18, Issue 2, Pages (January 2008)
Marie-Thérèse Leccia  Journal of Investigative Dermatology 
Lipid Rafts Establish Calcium Waves in Hepatocytes
Volume 7, Issue 6, Pages (December 2004)
Cell-Cell Adhesion and RhoA-Mediated Actin Polymerization are Independent Phenomena in Microtubule Disrupted Keratinocytes  Sun-Ho Kee, Shyh-Ing Jang,
Volume 11, Issue 18, Pages (September 2001)
Volume 14, Issue 1, Pages (January 2004)
Segregation of COPI-rich and anterograde-cargo-rich domains in endoplasmic- reticulum-to-Golgi transport complexes  David T. Shima, Suzie J. Scales, Thomas.
S100A7 (Psoriasin) Interacts with Epidermal Fatty Acid Binding Protein and Localizes in Focal Adhesion-Like Structures in Cultured Keratinocytes  Monica.
A PtdIns(3)P-specific probe cycles on and off host cell membranes during Salmonella invasion of mammalian cells  K. Pattni, M. Jepson, H. Stenmark, G.
X-Chromosome Inactivation Patterns Are Unbalanced and Affect the Phenotypic Outcome in a Mouse Model of Rett Syndrome  Juan I. Young, Huda Y. Zoghbi 
The Role of the Calcium Sensing Receptor in Regulating Intracellular Calcium Handling in Human Epidermal Keratinocytes  Chia-Ling Tu, Wenhan Chang, Daniel.
Contribution of Src-FAK signaling to the induction of connective tissue growth factor in renal fibroblasts  A. Graness, I. Cicha, M. Goppelt-Struebe 
Rapid Actin-Based Plasticity in Dendritic Spines
Expression of T-Cadherin in Basal Keratinocytes of Skin
Interactions of Human Myosin Va Isoforms, Endogenously Expressed in Human Melanocytes, Are Tightly Regulated by the Tail Domain  Wendy Westbroek, Jo Lambert,
Volume 15, Issue 7, Pages (April 2005)
Guidance of Cell Migration by Substrate Dimension
Septins Regulate Actin Organization and Cell-Cycle Arrest through Nuclear Accumulation of NCK Mediated by SOCS7  Brandon E. Kremer, Laura A. Adang, Ian.
Amanda Helip-Wooley, Wendy Westbroek, Heidi M
Intranuclear Localization of Insulin-Like Growth Factor Binding Protein-3 (IGFBP-3) During Cell Division in Human Keratinocytes  Christopher J. Wraight,
Effects of tumor necrosis factor-alpha on porcine oocyte meiosis progression, spindle organization, and chromosome alignment  Cai-Hong Ma, M.D., Li-Ying.
Yuri Oleynikov, Robert H. Singer  Current Biology 
Differential Effect of Wounding on Actin and its Associated Proteins, Paxillin and Gelsolin, in Fetal Skin Explants  Allison J. Cowin, Nicholas Hatzirodos,
Volume 138, Issue 4, Pages (August 2009)
Volume 131, Issue 3, Pages (September 2006)
Cadherin-Directed Actin Assembly
Rab11 Is Associated with Epidermal Lamellar Granules
Volume 68, Issue 6, Pages (December 2005)
Myosin V Colocalizes with Melanosomes and Subcortical Actin Bundles Not Associated with Stress Fibers in Human Epidermal Melanocytes  Jo Lambert, Yves.
Volume 11, Issue 13, Pages (July 2001)
Oxysterol-Binding Protein-Related Protein 1L Regulates Cholesterol Egress from the Endo-Lysosomal System  Kexin Zhao, Neale D. Ridgway  Cell Reports 
Drug-Induced Changes of Cytoskeletal Structure and Mechanics in Fibroblasts: An Atomic Force Microscopy Study  Christian Rotsch, Manfred Radmacher  Biophysical.
Yann Leverrier, Anne J. Ridley  Current Biology 
The bacterial actin nucleator protein ActA of Listeria monocytogenes contains multiple binding sites for host microfilament proteins  Susanne Pistor,
Volume 11, Issue 13, Pages (July 2001)
Rac1 Mediates Dendrite Formation in Response to Melanocyte Stimulating Hormone and Ultraviolet Light in a Murine Melanoma Model  Glynis A. Scott, Linda.
EGF induces HPSE nucleolar localization in human BMBC cells.
Yun-Gui Yang, Tomas Lindahl, Deborah E. Barnes  Cell 
Presentation transcript:

The Deficiency of PIP2 5-Phosphatase in Lowe Syndrome Affects Actin Polymerization  Sharon F. Suchy, Robert L. Nussbaum  The American Journal of Human Genetics  Volume 71, Issue 6, Pages 1420-1427 (December 2002) DOI: 10.1086/344517 Copyright © 2002 The American Society of Human Genetics Terms and Conditions

Figure 1 Actin stress-fiber staining is reduced in Lowe fibroblasts. F-actin staining was assessed in Lowe and normal fibroblasts, using Alexa 488 conjugated phalloidin to detect F-actin. Stained cells were classified into one of four types of actin staining: examples of each type (a–d), are shown in normal cells. Scale bar=0.016 μm. The histograms (e and f) show the results of two experiments with different Lowe and normal fibroblast lines, in which 100 (e) and 220 cells (f) of each type were scored by an observer masked to the genotype. Lowe syndrome fibroblasts had fewer cells with type a and b staining patterns and more cells with type c and d staining patterns, as compared with control fibroblasts (χ2=38.00, df 7, P<10−11). These results demonstrate that Lowe fibroblasts have a reduced number of long actin stress fibers. The American Journal of Human Genetics 2002 71, 1420-1427DOI: (10.1086/344517) Copyright © 2002 The American Society of Human Genetics Terms and Conditions

Figure 2 The F-actin staining pattern of Lowe cells shows a punctate pattern in the center of the cell, essentially absent from control cells. Lowe (a) and normal (b) fibroblasts were stained for F-actin using a fluorescent conjugated phalloidin, as described in figure 1. Scale bar=0.02 μm. The American Journal of Human Genetics 2002 71, 1420-1427DOI: (10.1086/344517) Copyright © 2002 The American Society of Human Genetics Terms and Conditions

Figure 3 Lowe fibroblasts had a more rapid response than controls to the actin depolymerizing agents, cytochalasin D and latrunculin A. Normal (a–c) and two Lowe fibroblast lines (d–f andg–i) were stained for F-actin using fluorescent conjugated phalloidin, as described in figure 1. Fibroblasts are shown untreated (a, d, and g) or incubated 5 min with 1 μmol cytochalasin D (b, e, and h) or with 0.24 μmol latrunculin A (c, f, andi). Lowe cells (e, f, h, and i) showed a greater response to depolymerizing agents than did control cells (b and c). Scale bar=0.02 μm. The American Journal of Human Genetics 2002 71, 1420-1427DOI: (10.1086/344517) Copyright © 2002 The American Society of Human Genetics Terms and Conditions

Figure 4 Gelsolin staining is altered in Lowe fibroblasts. Control and two Lowe fibroblast cultures were fixed, permeabilized, and immunostained with a monoclonal antibody to gelsolin, followed by an Alexa 594 conjugated secondary antibody. In control cells (a), gelsolin staining was distributed along stress fibers. In Lowe fibroblasts (b and c), there was an accumulation of punctate staining in the perinuclear area that was virtually absent in normal fibroblasts. Scale bar=0.02 μm. The American Journal of Human Genetics 2002 71, 1420-1427DOI: (10.1086/344517) Copyright © 2002 The American Society of Human Genetics Terms and Conditions

Figure 5 Punctate gelsolin staining in Lowe fibroblasts coincides with the punctate F-actin filament staining. Lowe fibroblasts were fixed and incubated with a monoclonal antibody to gelsolin followed by an Alexa 594 conjugated anti-rabbit secondary antibody, along with Alexa 488 conjugated phalloidin. Fluorescence was visualized by confocal microscopy. The punctate gelsolin staining (a) and punctate F-actin staining (b) coincided, as shown in the merged image (c). Scale bar=0.02 μm. The American Journal of Human Genetics 2002 71, 1420-1427DOI: (10.1086/344517) Copyright © 2002 The American Society of Human Genetics Terms and Conditions

Figure 6 Distribution of the punctate gelsolin staining does not colocalize with the TGN but does overlap with that of an ER marker. Lowe fibroblasts (a–c) were fixed and incubated with a monoclonal antibody to gelsolin and a polyclonal antibody to the mannose 6-phosphate receptor (M6-PR), followed by an Alexa 594 conjugated anti-mouse secondary antibody and an Alexa 488 conjugated anti-rabbit secondary antibody. The gelsolin staining (a) did not overlap with that of the TGN marker, M6-PR (b), as seen in the merged image (c). Scale bar=0.06 μm. The localization of the punctate gelsolin staining was further studied (d–f) by double-labeling Lowe fibroblasts, as described above, with anti-gelsolin antibody and with Alexa 488 conjugated concanavalin A, a probe for the endoplasmic reticulum. Fluorescence was visualized by confocal microscopy (d–f), scale bar=0.063 μm. The gelsolin staining (d) and the concanavalin A staining (e) partially overlapped, as shown in the merged image (f). The American Journal of Human Genetics 2002 71, 1420-1427DOI: (10.1086/344517) Copyright © 2002 The American Society of Human Genetics Terms and Conditions

Figure 7 Lowe cells transfected with ocrl1 cDNA are lacking the punctate gelsolin staining seen in most untransfected fibroblasts. Images show gelsolin staining of transfected (arrows) and untransfected cells. Following transfection, cells were plated, fixed, and double-labeled with an antibody to ocrl1, to identify transfected cells, and with gelsolin antibody. Scale bar=0.05 μm. The American Journal of Human Genetics 2002 71, 1420-1427DOI: (10.1086/344517) Copyright © 2002 The American Society of Human Genetics Terms and Conditions

Figure 8 Immunofluorescence staining of α-actinin is altered in Lowe fibroblasts. Control and two Lowe fibroblast cultures were fixed, permeabilized, and immunostained with a monoclonal antibody to α-actinin followed by an Alexa 488 conjugated secondary antibody. In control cells (a), α-actinin staining was present at focal adhesions and along stress fibers. In Lowe fibroblasts (b and c), there was an accumulation of punctate staining in the perinuclear area that was virtually absent in normal fibroblasts. Scale bar=0.05 μm. The American Journal of Human Genetics 2002 71, 1420-1427DOI: (10.1086/344517) Copyright © 2002 The American Society of Human Genetics Terms and Conditions