Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Actin Polymerization01:42

Actin Polymerization

Actin polymerization occurs through the head-to-tail association of binding sites on monomeric actin or G-actin to form filamentous or F-actin. The polymerization can be divided into three phases ̶  nucleation, elongation, and steady-state phase.
The nucleation phase involves forming a stable nucleus consisting of three actin monomers to form a new actin filament. Actin-binding proteins such as formins and Arp2/3 complex help filament growth post-nucleation. The Formins form straight actin...
Actin Filament Depolymerization01:19

Actin Filament Depolymerization

Actin filaments (F-actin) are composed of actin subunits. The dissociation of actin monomers can occur from either end of F-actin. The rate of dissociation is faster from the minus-end or the pointed end, where the actin subunits exist with a bound ADP, together known as ADP-actin. The depolymerization of F-actin is aided by proteins, including the actin-depolymerizing factor (ADF) and cofilin family of proteins, gelsolin, and glia maturation factor (GMF).
In F-actin, the ADF/cofilin proteins...
The Role of Actin and Myosin in Non-muscle Cells01:10

The Role of Actin and Myosin in Non-muscle Cells

Actin and myosin or actomyosin filaments also play a significant role in cells other than those involved in muscle contraction (which occurs within the sarcomere of muscle cells). The mechanism of non-muscle cell contractile bundles was first observed in Dictyostelium and Acanthamoeba. In non-muscle cells, two bundles are commonly found: stress fibers and actomyosin adherence belts. These contractile bundles are smaller and less organized than the ones found in muscle cells. They  are held...
Mechanism of Filopodia Formation01:39

Mechanism of Filopodia Formation

Filopodia are thin, actin-rich cellular protrusions that play an important role in many fundamental cellular functions. They vary in their occurrence, length, and positioning in different cell types, suggesting their diverse roles.
Their main function is to guide migrating cells during normal tissue morphogenesis or cancer metastasis by recognizing and making initial contacts with the extracellular matrix. However, they can also act as stationary cell anchors or help to establish communication...
Cell Motility through Blebbing01:16

Cell Motility through Blebbing

Blebs are a type of membrane protrusion formed by the internal hydrostatic pressure of the cytoplasm. Blebs are observed in several cell types, including fibroblasts, immune cells, and single-celled organisms like the amoeba. The primary function of blebs is cell locomotion and apoptosis, but they are also found during necrosis and cell division. The life cycle of a bleb comprises an initiation phase followed by the expansion and retraction phases.
Blebbing Through the Matrix
In multicellular...
Calmodulin-dependent Signaling01:16

Calmodulin-dependent Signaling

Calmodulin (CaM) is a calcium-binding protein in eukaryotes that controls various calcium-regulated cellular processes. It has four calcium-binding sites that bind calcium to form the calcium-calmodulin ( Ca2+-CaM) complex. GPCR stimulation increases the calcium levels in the cells that bind to CaM and induces a conformational change.
The Ca2+-CaM complex does not have enzymatic activity by itself. Instead, the complex binds downstream target proteins, including membrane proteins or enzymes,...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Inhibitory crosstalk between ERK and AMPK in the growth and proliferation of cardiac fibroblasts.

Biochemical and biophysical research communications·2008
Same author

mTORC1 signaling requires proteasomal function and the involvement of CUL4-DDB1 ubiquitin E3 ligase.

Cell cycle (Georgetown, Tex.)·2008
Same author

Prospective study of liver transplant recipients with HCV infection: evidence for a causal relationship between HCV and insulin resistance.

Liver transplantation : official publication of the American Association for the Study of Liver Diseases and the International Liver Transplantation Society·2008
Same author

Quantitative gel electrophoresis: sources of variation.

Journal of proteome research·2008
Same author

Evidence that the Nijmegen breakage syndrome protein, an early sensor of double-strand DNA breaks (DSB), is involved in HIV-1 post-integration repair by recruiting the ataxia telangiectasia-mutated kinase in a process similar to, but distinct from, cellular DSB repair.

Virology journal·2008
Same author

[Inhibitory effects of Qushi Huayu Decoction on fatty deposition and tumor necrosis factor alpha secretion in HepG2 cells induced by free fatty acid].

Zhongguo Zhong xi yi jie he za zhi Zhongguo Zhongxiyi jiehe zazhi = Chinese journal of integrated traditional and Western medicine·2008

Related Experiment Video

Updated: Jun 14, 2026

Cytosolic Calcium Measurements in Renal Epithelial Cells by Flow Cytometry
10:24

Cytosolic Calcium Measurements in Renal Epithelial Cells by Flow Cytometry

Published on: October 28, 2014

Calyculin A-induced actin phosphorylation and depolymerization in renal epithelial cells.

Luo Gu1, Hui Zhang, Qi Chen

  • 1Department of Life Sciences, Indiana State University, Terre Haute, IN 47809, USA.

Cell Motility and the Cytoskeleton
|February 26, 2003
PubMed
Summary

Renal epithelial cells phosphorylate actin, altering their cytoskeleton. This actin phosphorylation, observed in LLC-PK1 cells, impacts cell structure and may regulate actin organization.

More Related Videos

Single-channel Analysis and Calcium Imaging in the Podocytes of the Freshly Isolated Glomeruli
12:19

Single-channel Analysis and Calcium Imaging in the Podocytes of the Freshly Isolated Glomeruli

Published on: June 27, 2015

Implementing Patch Clamp and Live Fluorescence Microscopy to Monitor Functional Properties of Freshly Isolated PKD Epithelium
08:46

Implementing Patch Clamp and Live Fluorescence Microscopy to Monitor Functional Properties of Freshly Isolated PKD Epithelium

Published on: September 1, 2015

Related Experiment Videos

Last Updated: Jun 14, 2026

Cytosolic Calcium Measurements in Renal Epithelial Cells by Flow Cytometry
10:24

Cytosolic Calcium Measurements in Renal Epithelial Cells by Flow Cytometry

Published on: October 28, 2014

Single-channel Analysis and Calcium Imaging in the Podocytes of the Freshly Isolated Glomeruli
12:19

Single-channel Analysis and Calcium Imaging in the Podocytes of the Freshly Isolated Glomeruli

Published on: June 27, 2015

Implementing Patch Clamp and Live Fluorescence Microscopy to Monitor Functional Properties of Freshly Isolated PKD Epithelium
08:46

Implementing Patch Clamp and Live Fluorescence Microscopy to Monitor Functional Properties of Freshly Isolated PKD Epithelium

Published on: September 1, 2015

Area of Science:

  • Cell Biology
  • Biochemistry
  • Renal Physiology

Background:

  • The actin cytoskeleton is crucial for renal epithelial cell structure and function.
  • Regulation of actin dynamics involves post-translational modifications like phosphorylation.
  • Understanding actin phosphorylation in renal cells is key to deciphering cellular processes.

Purpose of the Study:

  • To investigate actin phosphorylation in the LLC-PK1 renal epithelial cell line.
  • To determine the role of actin phosphorylation in regulating the actin cytoskeleton.
  • To identify conditions and factors involved in actin phosphorylation.

Main Methods:

  • In vitro phosphorylation assays using cell lysate, phosphatase inhibitors, and ATP.
  • Detection of phosphorylated actin and kinase activity in cytoskeletal fractions.
  • In vivo studies using LLC-PK1 cells treated with calyculin A and ATP depletion.
  • Analysis of actin cytoskeleton organization via F-actin staining and microscopy.

Main Results:

  • Actin serine phosphorylation was detected in vitro and in living LLC-PK1 cells.
  • Phosphorylated actin and actin kinase activity were localized to the cytoskeletal fraction.
  • Calyculin A induced actin phosphorylation, leading to actin depolymerization, loss of cell junctions, and cytoskeletal reorganization.
  • ATP depletion abolished actin phosphorylation, while staurosporine did not affect it.

Conclusions:

  • LLC-PK1 renal epithelial cells possess kinase and phosphatase activities capable of phosphorylating actin.
  • Actin phosphorylation is linked to significant alterations in the actin cytoskeleton.
  • Actin phosphorylation likely plays a regulatory role in the organization of the actin cytoskeleton within renal epithelial cells.