TY - JOUR
T1 - Imaging centrosomes and cilia in the mouse kidney
AU - Hoshi, Masato
AU - Wang, Jinzhi
AU - Jain, Sanjay
AU - Mahjoub, Moe R.
N1 - Publisher Copyright:
© 2015. Published by Elsevier Inc.
PY - 2015
Y1 - 2015
N2 - The centrosome and cilium are evolutionarily conserved components of the microtubule cytoskeleton, and act as a cellular signaling center that regulates the activity of numerous developmental signaling pathways. Several genetic syndromes, called the ciliopathies, are associated with defects in the structure or function of the centrosome-cilium complex. In the mammalian kidney, these organelles are found at the apical surface of renal epithelial cells lining the various segments of the nephron, where they relay information from the extracellular environment to the interior of the cell. Cilium-based signaling plays an important role in the development and homeostasis of mammalian kidneys, and ciliary dysfunction is implicated in the pathogenesis of cystic kidney disease. Given the importance of centrosomes and cilia in renal function, techniques used to visualize these organelles, analyze their composition, and test their functionality have become essential in many studies of kidney development and disease. Fluorescence microscopy is a powerful, widely used technique that has enhanced our understanding of molecular mechanisms that regulate the assembly, maintenance, and function of these organelles in various organs. Here, we present detailed steps for the isolation of kidneys from adult and embryonic mice, describe protocols to label centrosomes and cilia in renal tissues, and methods used to culture and image kidneys ex vivo.
AB - The centrosome and cilium are evolutionarily conserved components of the microtubule cytoskeleton, and act as a cellular signaling center that regulates the activity of numerous developmental signaling pathways. Several genetic syndromes, called the ciliopathies, are associated with defects in the structure or function of the centrosome-cilium complex. In the mammalian kidney, these organelles are found at the apical surface of renal epithelial cells lining the various segments of the nephron, where they relay information from the extracellular environment to the interior of the cell. Cilium-based signaling plays an important role in the development and homeostasis of mammalian kidneys, and ciliary dysfunction is implicated in the pathogenesis of cystic kidney disease. Given the importance of centrosomes and cilia in renal function, techniques used to visualize these organelles, analyze their composition, and test their functionality have become essential in many studies of kidney development and disease. Fluorescence microscopy is a powerful, widely used technique that has enhanced our understanding of molecular mechanisms that regulate the assembly, maintenance, and function of these organelles in various organs. Here, we present detailed steps for the isolation of kidneys from adult and embryonic mice, describe protocols to label centrosomes and cilia in renal tissues, and methods used to culture and image kidneys ex vivo.
KW - Centriole
KW - Centrosome
KW - Cilium
KW - Polycystic kidney disease
KW - Renal
UR - http://www.scopus.com/inward/record.url?scp=84955407695&partnerID=8YFLogxK
U2 - 10.1016/bs.mcb.2014.12.008
DO - 10.1016/bs.mcb.2014.12.008
M3 - Article
C2 - 25837383
AN - SCOPUS:84955407695
SN - 0091-679X
VL - 127
SP - 1
EP - 17
JO - Methods in cell biology
JF - Methods in cell biology
ER -