Journal: Investigative Ophthalmology & Visual Science

Article Title: Mice Deficient in TAZ (Wwtr1) Demonstrate Clinical Features of Late-Onset Fuchs’ Endothelial Corneal Dystrophy

doi: 10.1167/iovs.64.4.22

Figure Lengend Snippet: Wwtr1 deficient mice exhibit reduced corneal endothelial cell density (ECD) and abnormal morphology when compared with WT controls. ( A ) In vivo confocal microscopy revealed reduced ECD in both Wwtr1 +/ − and Wwtr1 − / − mice at 2 months compared with WT mice. Despite age-related corneal endothelial cell (CEnC) loss, Wwtr1 deficient mice continued to have further declines in ECD at 6 and 12 months of age. Additionally, CEnCs from Wwtr1 deficient mice demonstrated abnormal morphology, lacking the normal hexagonal shape and even distribution, and altered reflectivity when compared with WT mice. ( B ) Each dot represents the mean ECD from three representative regions in a single eye ( n ≥ 9 eyes in each group), horizonal line represents mean of the group and error bars reflect standard deviation. A 2-way ANOVA was performed to detect statistical differences, *, **, ***, **** represent P < 0.05, P < 0.01, P < 0.001, P < 0.0001, respectively. Scale bar equivalent to 50 µm.

Article Snippet: Fourier-domain optical coherence tomography (FD-OCT; RTVue 100, software version 6.1; Optovue, Inc., Fremont, CA, USA) with a CAM-S lens (3-mm scan length) and in vivo confocal microscopy (IVCM; Heidelberg Retinal Tomography 3 in conjunction with the Rostock Cornea Module; Heidelberg Engineering, Heidelberg, Germany) were utilized to measure central corneal thickness (CCT) and endothelial cell density (ECD), respectively, with three representative regions counted for each eye.

Techniques: In Vivo, Confocal Microscopy, Standard Deviation

Journal: Investigative Ophthalmology & Visual Science

Article Title: Mice Deficient in TAZ (Wwtr1) Demonstrate Clinical Features of Late-Onset Fuchs’ Endothelial Corneal Dystrophy

doi: 10.1167/iovs.64.4.22

Figure Lengend Snippet: Wwtr1 deficient mice demonstrate abnormal tissue biomechanics of Descemet's membrane and irregular protein localization in corneal endothelial cells (CEnCs) compared with WT controls. ( A ) Atomic force microscopy (AFM) performed on Descemet's membrane revealed decreased elastic modulus (softer) in Wwtr1 deficient mice, with Wwtr1 − / − mice having the lowest measurement, followed by Wwtr1 +/ − mice and finally WT mice (stiffest). Each dot represents a single eye ( n ≥ 9 eyes in each group with each dot representing an average elastic modulus over five locations and five force curves were generated for each location), horizonal line represents mean of the group and error bars reflect standard deviation. A 2-way ANOVA was performed to detect statistical differences, *, **, ***, **** represent P < 0.05, P < 0.01, P < 0.001, P < 0.0001, respectively. ( B , C ) Representative CEnC flatmount preparations from WT, Wwtr1 +/ − , and Wwtr1 − / − mice at 2, 6, and 12 months of age, staining for Na/K-ATPase ( B , green ) and ZO-1 ( C , red ), and nuclei (DAPI, blue ). Both WT and Wwtr1 +/ − demonstrated discrete localization of Na/K-ATPase and ZO-1 to the cell periphery, whereas CEnCs from Wwtr1 − / − mice had diffuse and lobulated staining for Na/K-ATPase and disrupted to absent ZO-1 staining. Scale bar equivalent to 20 µm.

Article Snippet: Fourier-domain optical coherence tomography (FD-OCT; RTVue 100, software version 6.1; Optovue, Inc., Fremont, CA, USA) with a CAM-S lens (3-mm scan length) and in vivo confocal microscopy (IVCM; Heidelberg Retinal Tomography 3 in conjunction with the Rostock Cornea Module; Heidelberg Engineering, Heidelberg, Germany) were utilized to measure central corneal thickness (CCT) and endothelial cell density (ECD), respectively, with three representative regions counted for each eye.

Techniques: Membrane, Microscopy, Generated, Standard Deviation, Staining

Journal: Investigative Ophthalmology & Visual Science

Article Title: Mice Deficient in TAZ (Wwtr1) Demonstrate Clinical Features of Late-Onset Fuchs’ Endothelial Corneal Dystrophy

doi: 10.1167/iovs.64.4.22

Figure Lengend Snippet: Wwtr1 deficient mice demonstrated impaired corneal endothelial wound healing after phototherapeutic keratectomy (PTK). An endothelial wound was created with an excimer laser (20 µm depth) in 6-month-old WT ( n = 6) and Wwtr1 +/− ( n = 6) mice. Optical coherence tomography (OCT) ( A, B ), slit lamp biomicroscopy ( C ), and in vivo confocal microscopy (IVCM) ( D ) were performed on post-injury days 1, 3, 5, 7, 10, and 14. Central corneal thickness (CCT) measured with OCT did not show statistical difference between the two genotypes ( B ), however, CCT reached its maximal thickness on post-injury days (PIDs) 7 to 10 and corneal edema was more marked on PID 10 in Wwtr1 +/− mice ( C ). IVCM revealed the regenerating course of injured endothelial cells throughout the study period ( D ). Following euthanasia on PID 14, endothelial wholemounts were stained with alizarin red ( E ), and endothelial cell density (ECD) was measured. ECD was significantly reduced in Wwtr1 +/− mice versus WT, suggesting impaired endothelial healing in Wwtr1 deficient mice. CCT was analyzed using a 2-way ANOVA, ECD was analyzed using a Mann-Whitney U test, * P < 0.05. OCT scale bars equivalent to 250 µm, IVCM and Alizarin red microscopy scale bars equivalent to 50 µm.

Article Snippet: Fourier-domain optical coherence tomography (FD-OCT; RTVue 100, software version 6.1; Optovue, Inc., Fremont, CA, USA) with a CAM-S lens (3-mm scan length) and in vivo confocal microscopy (IVCM; Heidelberg Retinal Tomography 3 in conjunction with the Rostock Cornea Module; Heidelberg Engineering, Heidelberg, Germany) were utilized to measure central corneal thickness (CCT) and endothelial cell density (ECD), respectively, with three representative regions counted for each eye.

Techniques: Tomography, In Vivo, Confocal Microscopy, Staining, MANN-WHITNEY, Microscopy