Epithelial cells are typically arranged in a honeycomb-like pattern, minimizing their cell-cell contact areas, which suggests that some tension operates for shaping of the cell boundaries. However, the molecular mechanisms that generate such tension remain unknown. We found that Tuba, which is a Cdc42-specific GEF, was concentrated at the apical-most region of cell junctions in simple epithelia via its interaction with ZO-1. RNAi-mediated depletion of Tuba altered the geometrical configuration of cell junctions, resulting in a curved and slack appearance. At the subcellular level, Tuba inactivation modified the assembly pattern of junctional F-actin and E-cadherin. Tuba RNAi also retarded cell junction formation in calcium-switch experiments. Suppression of Cdc42 activity or depletion of N-WASP, which is an effector of Cdc42, mimicked the effects of Tuba depletion. Conversely, overexpression of dominant-active Cdc42 or N-WASP enhanced the junction formation of Tuba-depleted cells. These results suggest that Tuba controls the shaping of cell junctions through the local activation of Cdc42 and its effectors.
Download full-text PDF |
Source |
---|---|
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2064505 | PMC |
http://dx.doi.org/10.1083/jcb.200605012 | DOI Listing |
Methods Mol Biol
January 2025
Quantum-Si, Guilford, CT, USA.
Single-molecule fluorescence resonance energy transfer (smFRET) is a powerful technique for studying the structural dynamics of protein molecules or detecting interactions between protein molecules in real time. Due to the high sensitivity in spatial and temporal resolution, smFRET can decipher sub-populations within heterogeneous native state conformations, which are generally lost in traditional measurements due to ensemble averaging. In addition, the single-molecule reconstitution allows protein molecules to be observed for an extensive period of time and can recapitulate the geometry of the cellular environment to retain biological function.
View Article and Find Full Text PDFJ Vis Exp
December 2024
Department of Pharmacology, School of Medicine, Ajou University; 3D Immune System Imaging Core Center, Ajou University;
Technical hurdles in a culture of epithelial cells include dedifferentiation and loss of function. Biomimetic three-dimensional (3D) cell culture methods can enhance cell culture efficiency. This study introduces an advanced two-layered culture system intended to cultivate epithelial cells as tissue-like layers with the culture of fibroblasts within a 3D environment.
View Article and Find Full Text PDFActa Biomater
January 2025
Ophthalmology Department, The First Affiliated Hospital of Jinan University, Guangzhou, China; Key Laboratory for Regenerative Medicine of Ministry of Education, Jinan University, Guangzhou, China; Institute of Ophthalmology, Medical College, Jinan University, Guangzhou, China; Aier School of Ophthalmology, Central South University, Changsha, China. Electronic address:
The shortage of corneal donors and the limitations in tissue engineering grafts, such as biocompatibility and mechanical properties, pose significant challenges in corneal transplantation. Here, for the first time, we investigate the effect of Rho kinase inhibitor Y-27632 and a dual media culture approach, including proliferative media (M1) and stabilizing media (M2), on rabbit limbal epithelial stem cells (LESCs), aiming to explore the feasibility of constructing corneal cell sheets in vitro through auto-bioprinting and assessing their corneal wound healing capacity in vivo. Y-27632 has primarily demonstrated significantly enhanced LESCs growth, proliferation, and reduced apoptosis.
View Article and Find Full Text PDFBurns
December 2024
Lee Kong Chian School of Medicine, Nanyang Technological University Singapore, Clinical Sciences Building, 11, Mandalay Road, 308232, Singapore; Skin Research Institute Singapore, Level 17, Clinical Sciences Building, 11, Mandalay Road, 308232, Singapore; National Skin Centre Singapore, 1 Mandalay Rd, 308205, Singapore. Electronic address:
Burns are dynamic injuries characterized by an initial zone of necrosis that progresses to compromise surrounding tissue. Acute inflammation and cell death are two main factors contributing to burn progression. These processes are modulated by Connexin43 (Cx43) hemichannels and gap junctions in burns and chronic wounds.
View Article and Find Full Text PDFInt J Mol Sci
January 2025
Vavilov Institute of General Genetics Russian Academy of Sciences, 119333 Moscow, Russia.
The size of viral genomes is limited, thus the majority of encoded proteins possess multiple functions. The main function of tobamoviral movement protein (MP) is to perform plasmodesmata gating and mediate intercellular transport of the viral RNA. MP is a remarkable example of a protein that, in addition to the initially discovered and most obvious function, carries out numerous activities that are important both for the manifestation of its key function and for successful and productive infection in general.
View Article and Find Full Text PDFEnter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!