scholarly journals Inhibitory Effects of Eucalyptol on Diabetes-associated Dysfunction of Actin Cytoskeleton and Focal Adhesion Formation in Kidney Podocytes (P06-011-19)

2019 ◽  
Vol 3 (Supplement_1) ◽  
Author(s):  
Dongyeon KIM ◽  
Young-Hee Kang
Keyword(s):  
Actin Cytoskeleton ◽  
Focal Adhesion ◽  
Adhesion Formation ◽  
Kidney Tissue ◽  
Cytoskeletal Proteins ◽  
Rhodamine Phalloidin ◽  
Adhesion Proteins ◽  
Tissue Levels ◽  
Focal Adhesion Proteins ◽  
Focal Adhesion Formation

Abstract Objectives Chronic hyperglycemia causes glomerular podocyte damage that can result in glomerular focal adhesion and cytoskeleton rearrangement. Eucalyptol (1,8-cineole) is a natural organic essential oil and a monoterpenoid with anti-inflammatory and antioxidant properties. Methods Immotalized mouse podocytes were incubated in media containing 33 mM glucose for 4 days in the presence of 1–20 μM eucalyptol. Antibodies of F-actin, ezrin, Arp2/3, cortactin, paxillin, vinculin, talin, and FAK were used for Western blot analysis. Podocytes were stained with rhodamine-phalloidin to stain actin filaments. The db/db mice were orally administrated with 10 mg/kg eucalyptol. Kidney tissue extracts were prepared for Western blotting and immunohistochemically stained. Results Glucose suppressed the induction of the cytoskeletal proteins responsible for the maintenance of podocyte cytoskeleton structural integrity. However, eucalyptol prompted such reduction in diabetic podocytes. Eucalyptol enhanced rhodamine-phalloidin-red staining of podocyte F-actin diminished in glucose-exposed podocytes. In addition, the tissue levels of the cytoskeletal proteins were reduced in diabetic kidneys. Oral administration of eucalyptol to db/db mice augmented the kidney tissue levels of cytoskeletal proteins and boosted glomerular level of F-actin. On the other hand, the induction of focal adhesion proteins was dampened in glucose-loaded podocytes, while the activation of these proteins inductions were highly elevated. The presence of eucalyptol reversed the aforementioned effects of focal adhesion proteins in diabetic podocytes. Furthermore, eucalyptol enhanced the decreased levels of the focal adhesion proteins in diabetic kidneys. Conclusions These results demonstrated that eucalyptol ameliorated actin cytoskeleton integrity and focal adhesion formation in diabetic kidneys. Therefore, eucalyptol may be a potent renoprotective agent counteracting diabetes-associated podocyte detachment and disruption of focal adhesion proteins. Funding Sources This work was supported by the National Research Foundation of Korea (NRF) grants funded by the Korea government (MEST) (NRF-2017R1A6A3A04011473).

scholarly journals SPEG binds with desmin and its deficiency causes defects in triad and focal adhesion proteins

2020 ◽  
Author(s):  
Shiyu Luo ◽  
Qifei Li ◽  
Jasmine Lin ◽  
Quinn Murphy ◽  
Isabelle Marty ◽  
...  
Keyword(s):  
Focal Adhesion ◽  
Skeletal Muscles ◽  
Striated Muscle ◽  
Calcium Handling ◽  
Intermediate Filament Protein ◽  
Β1 Integrin ◽  
Adhesion Proteins ◽  
Focal Adhesion Proteins ◽  
Early Endosomes

Abstract SPEG, a member of the myosin light chain kinase family, is localized at the level of triad surrounding myofibrils in skeletal muscles. In humans, SPEG mutations are associated with centronuclear myopathy and cardiomyopathy. Using a striated muscle specific Speg-knockout (KO) mouse model, we have previously shown that SPEG is critical for triad maintenance and calcium handling. Here we further examined the molecular function of SPEG and characterized the effects of SPEG deficiency on triad and focal adhesion proteins. We used yeast two-hybrid assay, and identified desmin, an intermediate filament protein, to interact with SPEG and confirmed this interaction by co-immunoprecipitation. Using domain-mapping assay, we defined that Ig-like and fibronectin III domains of SPEG interact with rod domain of desmin. In skeletal muscles, SPEG depletion leads to desmin aggregates in vivo and a shift in desmin equilibrium from soluble to insoluble fraction. We also profiled the expression and localization of triadic proteins in Speg-KO mice using western blot and immunofluorescence. The amounts of RyR1 and triadin were markedly reduced, whereas DHPRα1, SERCA1, and triadin were abnormally accumulated in discrete areas of Speg-KO myofibers. In addition, Speg-KO muscles exhibited internalized vinculin and β1 integrin, both of which are critical components of the focal adhesion complex. Further, β1 integrin was abnormally accumulated in early endosomes of Speg-KO myofibers. These results demonstrate that SPEG-deficient skeletal muscles exhibit several pathological features similar to those seen in MTM1 deficiency. Defects of shared cellular pathways may underlie these structural and functional abnormalities in both types of diseases.

The Kindlin protein family: new members to the club of focal adhesion proteins

2009 ◽  
Vol 19 (10) ◽  
pp. 504-513 ◽  
Author(s):  
Alexander Meves ◽  
Christopher Stremmel ◽  
Kay Gottschalk ◽  
Reinhard Fässler
Keyword(s):  
Focal Adhesion ◽  
Protein Family ◽  
New Members ◽  
Adhesion Proteins ◽  
Focal Adhesion Proteins

Abstract 110: Thrombospondin Type-1 Domain-Containing Protein 1 (THSD1), an Intracranial Aneurysm Susceptibility Gene, Mediates Cell Adhesion in Human Umbilical Vein Endothelial Cells

Stroke ◽  
2015 ◽  
Vol 46 (suppl_1) ◽  
Author(s):  
Xiaoqian Fang ◽  
Dong H Kim ◽  
Teresa Santiago-Sim
Keyword(s):  
Endothelial Cells ◽  
Cell Adhesion ◽  
Focal Adhesion ◽  
Umbilical Vein ◽  
Adhesion Formation ◽  
Wild Type ◽  
Focal Adhesion Formation ◽  
Umbilical Vein Endothelial Cells

Introduction: An intracranial aneurysm (IA) is a weak spot in cerebral blood vessel wall that can lead to its abnormal bulging. Previously, we reported that mutations in THSD1 , encoding thrombospondin type-1 domain-containing protein 1, are associated with IA in a subset of patients. THSD1 is a transmembrane molecule with a thrombospondin type-1 repeat (TSR). Proteins with TSR domain have been implicated in a variety of processes including regulation of matrix organization, cell adhesion and migration. We have shown that in mouse brain Thsd1 is expressed in endothelial cells. Hypothesis: THSD1 plays an important role in maintaining the integrity of the endothelium by promoting adhesion of endothelial cells to the underlying basement membrane. Methods: Human umbilical vein endothelial cells are used to investigate the role of THSD1 in vitro . THSD1 expression was knocked-down by RNA interference. Cell adhesion assay was done on collagen I-coated plates and focal adhesion formation was visualized using immunofluorescence by paxillin and phosphorylated focal adhesion kinase (pFAK) staining. THSD1 re-expression is accomplished by transfection with a pCR3.1-THSD1-encoding plasmid. Results: Knockdown of THSD1 caused striking change in cell morphology and size. Compared to control siRNA-treated cells that exhibited typical cobblestone morphology, THSD1 knockdown cells were narrow and elongated, and were significantly smaller ( p <0.01). Cell adherence to collagen I-coated plates was also attenuated in THSD1 knockdown cells ( p <0.01). Consistent with this finding is the observation that the number and size of focal adhesions, based on paxillin and pFAK staining, were significantly reduced after THSD1 knockdown ( p <0.01). These defects in cell adhesion and focal adhesion formation were rescued by re-expression of wild type THSD1 ( p <0.05). In contrast, initial studies indicate that expression of mutated versions of THSD1 as seen in human patients (L5F, R450*, E466G, P639L) could not restore cell adhesion and focal adhesion formation to wild type levels. Conclusions: Our studies provide evidence for a role of THSD1 and THSD1 mutations in endothelial cell adhesion and suggest a possible mechanism underlying THSD1 -mediated aneurysm disease.

Nanoporosity Stimulates Cell Spreading and Focal Adhesion Formation in Cells with Mutated Paxillin

2020 ◽  
Vol 12 (13) ◽  
pp. 14924-14932
Author(s):  
Dainelys Guadarrama Bello ◽  
Aurélien Fouillen ◽  
Antonella Badia ◽  
Antonio Nanci
Keyword(s):  
Focal Adhesion ◽  
Adhesion Formation ◽  
Cell Spreading ◽  
Focal Adhesion Formation ◽  
In Cells
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