Volume 196, Issue 1, Pages (July 2016)

Slides:



Advertisements
Similar presentations
Volume 184, Issue 4, Pages (October 2010)
Advertisements

A Rat Model for Choroidal Neovascularization Using Subretinal Lipid Hydroperoxide Injection  Takayuki Baba, Imran A. Bhutto, Carol Merges, Rhonda Grebe,
From: Engraftment of Adult Neural Progenitor Cells Transplanted to Rat Retina Injured by Transient Ischemia Invest. Ophthalmol. Vis. Sci ;44(7):
Volume 9, Issue 5, Pages (November 2017)
Involvement of Wnt Signaling in Dermal Fibroblasts
Bioengineered vascular access maintains structural integrity in response to arteriovenous flow and repeated needle puncture  Bryan W. Tillman, MD, PhD,
Wound healing around and within saphenous vein bypass grafts
Studies on the Growth of Cells
Volume 10, Issue 1, Pages (January 2012)
Supplementary figure 1 D
Volume 199, Issue 1, Pages (January 2018)
Volume 134, Issue 5, Pages (May 2008)
Volume 65, Issue 1, Pages (January 2004)
Proliferation, Cell Cycle Exit, and Onset of Terminal Differentiation in Cultured Keratinocytes: Pre-Programmed Pathways in Control of C-Myc and Notch1.
Ken Suzuki, MD, PhD, Nigel J. Brand, PhD, Sean Allen, PhD, Mahboob A
Epstein–Barr Virus Infection Induces Aberrant TLR Activation Pathway and Fibroblast– Myofibroblast Conversion in Scleroderma  Antonella Farina, Mara Cirone,
Volume 132, Issue 2, Pages (February 2007)
VEGF Gene Delivery to Muscle
Volume 74, Issue 7, Pages (October 2008)
Volume 4, Issue 3, Pages (March 2015)
Volume 8, Issue 6, Pages (June 2017)
Vida K. Stark, MS, Thomas F. Warner, MD, John R. Hoch, MD 
Approach for the Derivation of Melanocytes from Induced Pluripotent Stem Cells  Tamihiro Kawakami, Tatsuro Okano, Sora Takeuchi, Kayoko Osumi, Yoshinao.
Protease-Activated Receptor-2 (PAR-2) Expression in Human Fibroblasts is Regulated by Growth Factors and Extracellular Matrix  Barry L. Gruber, Mary J.
Homing of intravenously infused embryonic stem cell-derived cells to injured hearts after myocardial infarction  Jiang-Yong Min, MD, Xuling Huang, MD,
Loss of EPC-1/PEDF Expression During Skin Aging In Vivo
Calcium Ion Gradients and Dynamics in Cultured Skin Slices of Rat Hindpaw in Response to Stimulation with ATP  Moe Tsutsumi, Sumiko Denda, Kaori Inoue,
Volume 18, Issue 2, Pages (February 2016)
Volume 3, Issue 2, Pages (August 2014)
The Spatial and Temporal Expression Patterns of Integrin α9β1 and One of Its Ligands, the EIIIA Segment of Fibronectin, in Cutaneous Wound Healing  Purva.
Scanning Near-Field Fluorescence Resonance Energy Transfer Microscopy
Volume 14, Issue 1, Pages (July 2011)
Volume 34, Issue 5, Pages (May 2011)
Volume 84, Issue 4, Pages (April 2003)
Volume 36, Issue 2, Pages (January 2016)
Volume 10, Issue 2, Pages (February 2018)
Lineage Tracing Using Cux2-Cre and Cux2-CreERT2 Mice
Volume 10, Issue 5, Pages (May 2012)
Calmodulin-Like Protein Upregulates Myosin-10 in Human Keratinocytes and Is Regulated during Epidermal Wound Healing In Vivo  Richard D. Bennett, Amy.
Definitive Hematopoiesis Requires the Mixed-Lineage Leukemia Gene
Volume 41, Issue 2, Pages (August 2014)
A Murine Living Skin Equivalent Amenable to Live-Cell Imaging: Analysis of the Roles of Connexins in the Epidermis  Eve E. Kandyba, Malcolm B. Hodgins,
Volume 61, Issue 6, Pages (June 2002)
Response Diversity and the Timing of Progenitor Cell Maturation Are Regulated by Developmental Changes in EGFR Expression in the Cortex  Robert C Burrows,
Volume 17, Issue 6, Pages (June 2009)
Volume 8, Issue 2, Pages (February 2017)
Derivation and FACS-Mediated Purification of PAX3+/PAX7+ Skeletal Muscle Precursors from Human Pluripotent Stem Cells  Bianca Borchin, Joseph Chen, Tiziano.
Molecular and Fluorescence In Situ Hybridization Characterization of the Breakpoints in 46 Large Supernumerary Marker 15 Chromosomes Reveals an Unexpected.
Role of VEGF-A in Vascularization of Pancreatic Islets
Local Arrangement of Fibronectin by Myofibroblasts Governs Peripheral Nuclear Positioning in Muscle Cells  William Roman, João P. Martins, Edgar R. Gomes 
Figure 1 VGCC antibody uptake in cerebellar slice culture
Volume 12, Issue 6, Pages (December 2005)
Susana Gomis-Rüth, Corette J. Wierenga, Frank Bradke  Current Biology 
Volume 4, Issue 3, Pages (March 2015)
Volume 38, Issue 4, Pages (April 2013)
John E. Olerud, Marcia L. Usui, Deniz Seckin, Diane S. Chiu, Claire L
Volume 99, Issue 8, Pages (October 2010)
Promotion of Human Dermal Fibroblast Migration, Matrix Remodelling and Modification of Fibroblast Morphology within a Novel 3D Model by Lucilia sericata.
Volume 10, Issue 3, Pages (March 2018)
Lentiviral Vector-Mediated Gene Transfer to Human Hair Follicles
Jun Asai, Hideya Takenaka, Norito Katoh, Saburo Kishimoto 
Alterations in Fibroblast α1β1 Integrin Collagen Receptor Expression in Keloids and Hypertrophic Scars  Greg Szulgit  Journal of Investigative Dermatology 
Organization of Stem Cells and Their Progeny in Human Epidermis
Volume 13, Issue 3, Pages (March 2006)
James Gailit, Mary J. Marchese, Richard R. Kew, Barry L. Gruber 
Volume 6, Issue 4, Pages (July 2013)
Comparison of Mouse Matrix Metalloproteinase 13 Expression in Free-Electron Laser and Scalpel Incisions During Wound Healing  Nanjun Wu, E. Duco Jansen,
Y. Albert Pan, Joshua R. Sanes  Journal of Investigative Dermatology 
Marrow stromal cells for cellular cardiomyoplasty: Feasibility and potential clinical advantages  Jih-Shiuan Wang, MDa,b, Dominique Shum-Tim, MDa, Jacques.
Characterization of Epidermal Wound Healing in a Human Skin Organ Culture Model: Acceleration by Transplanted Keratinocytes1  Ingrid Moll, Pia Houdek,
Presentation transcript:

Volume 196, Issue 1, Pages 270-278 (July 2016) Fetal Rat Gubernaculum Mesenchymal Cells Adopt Myogenic and Myofibroblast-Like Phenotypes  Alan K. Robbins, Abigail B. Mateson, Ashutosh Khandha, Joan E. Pugarelli, Thomas S. Buchanan, Robert E. Akins, Julia Spencer Barthold  The Journal of Urology  Volume 196, Issue 1, Pages 270-278 (July 2016) DOI: 10.1016/j.juro.2015.12.081 Copyright © 2016 American Urological Association Education and Research, Inc. Terms and Conditions

Figure 1 Immunostaining of E17 (A and B), E19 (C) and E21 (D) rat GCC sections for desmin (green areas). Distal GCC is shown on left side and more proximal GCC is shown on right side of each frame. Differentiated muscle layers stained brightly for desmin. Single thin peripheral muscle layer was visible at E17 with opening present at GCC tip (left side) (A). Desmin expression was much lower in central mesenchymal core but enriched at tip. More proximal image of same section shows second, inner muscle layer developing in proximal half of GCC and enrichment of desmin positive mesenchymal cells between muscle layers as apparent myogenic zone (bracket) (B). Thickening of both muscle layers was visible at E19 (C). Further development by E21 obliterated space between muscle layers (top) (D). Scale bar indicates 200 (A and B) and 400 μm (C and D). The Journal of Urology 2016 196, 270-278DOI: (10.1016/j.juro.2015.12.081) Copyright © 2016 American Urological Association Education and Research, Inc. Terms and Conditions

Figure 2 Immunostaining of E17 (A), E19 (B) and E21 (C) GCC sections for αSMA (red areas) and myogenin (green areas). Sections are adjacent to those in figure 3. Distal GCC is on right side and more proximal GCC is on left side of each frame. Differentiated muscle layers stained brightly for αSMA. DAPI counterstaining (Merge) shows position of inner mesenchyme relative to muscle layers. At E17 cells with nuclear myogenin staining were primarily in but occasionally between differentiated muscle layers (A). At E19 additional myogenin positive cells were visible between developing muscle layers (right side) that defined myogenic zone (B). Scale bar (C) indicates 200 μm. The Journal of Urology 2016 196, 270-278DOI: (10.1016/j.juro.2015.12.081) Copyright © 2016 American Urological Association Education and Research, Inc. Terms and Conditions

Figure 3 Immunostaining of E17 (A), E19 (B) and E21 (C) GCC sections for desmin (green areas) and PAX7 (red areas). Sections are adjacent to those in figure 2. Distal GCC is on right side and more proximal GCC is on left side of each frame. Differentiated muscle layers stained brightly for desmin. DAPI counterstaining (Merge) shows position of inner mesenchyme relative to muscle layers. At E17 cells with nuclear PAX7 staining were primarily within but occasionally between differentiated muscle layers (A). At E19 additional PAX7 positive cells were visible between developing muscle layers (right side) that defined myogenic zone (B). Scale bar (C) indicates 200 μm. The Journal of Urology 2016 196, 270-278DOI: (10.1016/j.juro.2015.12.081) Copyright © 2016 American Urological Association Education and Research, Inc. Terms and Conditions

Figure 4 Immunostaining of E19 GCC sections for αSMA (red areas) (A) and PECAM/CD31 (green areas) (B) shows strong αSMA expression in differentiated muscle (left upper corner) and fainter mesenchymal expression in GCC core (right lower corner). αSMA and PECAM co-expression in blood vessels appeared yellow (Merge). Inset, higher magnification of PECAM negative region in core is enhanced to better visualize mesenchymal αSMA. Scale bar indicates 100 μm. The Journal of Urology 2016 196, 270-278DOI: (10.1016/j.juro.2015.12.081) Copyright © 2016 American Urological Association Education and Research, Inc. Terms and Conditions

Figure 5 CellTracker fluorescent staining (green areas) of mesothelial layer after 5-minute exposure of E17 GCC was followed by immediate fixation at 0 hours (A). Dye uptake by mesothelial cells outside peripheral muscle layer is delineated by MyHC immunostaining (red areas). Note evidence of labeled mesothelial cell (green areas and arrows) incorporation in GCC deeper muscle and mesenchymal regions after 5-minute CellTracker exposure followed by 24-hour organ culture (B). Scale bar indicates 100 μm. The Journal of Urology 2016 196, 270-278DOI: (10.1016/j.juro.2015.12.081) Copyright © 2016 American Urological Association Education and Research, Inc. Terms and Conditions

Figure 6 Low (A) and high (B) power phase contrast images reveal 3-day cultured intact E17 GCC grown on poly-L-lysine/laminin coated plates and immobilized in Matrigel. Progressive peripheral cell migration was observed throughout culture period from all GCC borders. Note anastomosing cellular architecture outside GCC (B). Cells migrating in Matrigel were imaged after fixation in situ followed by immunostaining for myogenic markers (C to E). Note ubiquitous αSMA expression (green areas) in migrating cells with nuclear localization of myogenin in occasional cells (red areas) (C). All migrating cells expressed desmin (green areas) (D). Subset also demonstrated nuclear localization of myogenic commitment marker PAX7 (red areas). All migrating cells expressed ITGA7 (green areas) (E). Differentiated muscle was visualized by MyHC expression (red areas), showing that some migrating cells formed striated myotubes. Scale bar indicates 200 μm (A to E). The Journal of Urology 2016 196, 270-278DOI: (10.1016/j.juro.2015.12.081) Copyright © 2016 American Urological Association Education and Research, Inc. Terms and Conditions

Figure 7 Phase contrast images show passage 4 or 6 GCC cells grown to confluence on collagen-I coated plates visualized after transfer to low serum (2%) medium. Note early appearance of cleared areas created by contraction of cellular monolayer beginning at day 4 in low serum medium (A). More generalized contraction of cellular monolayer created elongated 3D cellular ridges after additional 5-day observation together with medium changes (B). Reduced from ×20. The Journal of Urology 2016 196, 270-278DOI: (10.1016/j.juro.2015.12.081) Copyright © 2016 American Urological Association Education and Research, Inc. Terms and Conditions