Format

Send to

Choose Destination
Methods Cell Biol. 2019;153:205-229. doi: 10.1016/bs.mcb.2019.04.008. Epub 2019 May 17.

Analysis of primary cilia in renal tissue and cells.

Author information

1
Department of Anatomy and Cell Biology, The Jared Grantham Kidney Institute, University of Kansas Medical Center, Kansas City, KS, United States.
2
Department of Anatomy and Cell Biology, The Jared Grantham Kidney Institute, University of Kansas Medical Center, Kansas City, KS, United States. Electronic address: ptran@kumc.edu.

Abstract

Primary cilia are singular, sensory organelles that extend from the plasma membrane of most quiescent mammalian cells. These slender, microtubule-based organelles receive and transduce extracellular cues and regulate signaling pathways. Primary cilia are critical to the development and function of many tissue types, and mutation of ciliary genes causes multi-system disorders, termed ciliopathies. Notably, renal cystic disease is one of the most common clinical features of ciliopathies, highlighting a central role for primary cilia in the kidney. Additionally, acute kidney injury and chronic kidney disease are associated with altered primary cilia lengths on renal epithelial cells, suggesting ciliary dynamics and renal physiology are linked. Here we describe methods to examine primary cilia in kidney tissue and in cultured renal cells. We include immunofluorescence and scanning electron microscopy to determine ciliary localization of proteins and cilia structure. Further, we detail cellular assays to measure cilia assembly and disassembly, which regulate cilia length.

KEYWORDS:

Cilia length; Gene knock-down; Immunofluorescence; Primary cilia; Renal epithelial cells; Scanning electron microscopy

PMID:
31395380
DOI:
10.1016/bs.mcb.2019.04.008

Supplemental Content

Full text links

Icon for Elsevier Science
Loading ...
Support Center