TY - JOUR
T1 - Scanning electron microscopy of human islet cilia
AU - Polino, Alexander J.
AU - Sviben, Sanja
AU - Melena, Isabella
AU - Piston, David W.
AU - Hughes, Jing W.
N1 - Funding Information:
ACKNOWLEDGMENTS. We are indebted to Dr. Ursula Goodenough along with Dr. Bob Schmidt, Robyn Roth, and the WashU Cilia Group for help with data interpretation. We acknowledge the assistance of Drs. James Fitzpatrick and Praveen Krishnamoorthy at the Washington University Center for Cellular Imaging in electron and light microscopy studies, supported by Washington University School of Medicine, The Children’s Discovery Institute of Washington University and St. Louis Children’s Hospital (CDI-CORE-2015-505 and CDI-CORE-2019-813),the Foundation for Barnes-Jewish Hospital (3770) and the Washington University Diabetes Research Center (DK020579). Human pancreatic islets were provided by the NIDDK-funded Integrated IsletDistribution Program (IIDP) (RRID:SCR_014387) at City of Hope,NIH grant #2UC4DK098085 and the JDRF-funded IIDP Islet Award Initiative.A.J.P.is supported by the Diabetes and Related Metabolic Diseases training grant T32DK007120. Other funding included NIH grants DK115795 and DK127748 to J.W.H.,Doris Duke Charitable Foundation grant DDFRCS to J.W.H., and DK115972 and DK123301 to D.W.P. Graphics were created with BioRender.com.
Publisher Copyright:
Copyright © 2023 the Author(s). Published by PNAS.
PY - 2023/5/30
Y1 - 2023/5/30
N2 - Human islet primary cilia are vital glucose-regulating organelles whose structure remains uncharacterized. Scanning electron microscopy (SEM) is a useful technique for studying the surface morphology of membrane projections like cilia, but conventional sample preparation does not reveal the submembrane axonemal structure, which holds key implications for ciliary function. To overcome this challenge, we combined SEM with membrane-extraction techniques to examine primary cilia in native human islets. Our data show well-preserved cilia subdomains which demonstrate both expected and unexpected ultrastructural motifs. Morphometric features were quantified when possible, including axonemal length and diameter, microtubule conformations, and chirality. We further describe a ciliary ring, a structure that may be a specialization in human islets. Key findings are correlated with fluorescence microscopy and interpreted in the context of cilia function as a cellular sensor and communications locus in pancreatic islets.
AB - Human islet primary cilia are vital glucose-regulating organelles whose structure remains uncharacterized. Scanning electron microscopy (SEM) is a useful technique for studying the surface morphology of membrane projections like cilia, but conventional sample preparation does not reveal the submembrane axonemal structure, which holds key implications for ciliary function. To overcome this challenge, we combined SEM with membrane-extraction techniques to examine primary cilia in native human islets. Our data show well-preserved cilia subdomains which demonstrate both expected and unexpected ultrastructural motifs. Morphometric features were quantified when possible, including axonemal length and diameter, microtubule conformations, and chirality. We further describe a ciliary ring, a structure that may be a specialization in human islets. Key findings are correlated with fluorescence microscopy and interpreted in the context of cilia function as a cellular sensor and communications locus in pancreatic islets.
KW - human islets
KW - primary cilia
KW - scanning electron microscopy
UR - http://www.scopus.com/inward/record.url?scp=85159646820&partnerID=8YFLogxK
U2 - 10.1073/pnas.2302624120
DO - 10.1073/pnas.2302624120
M3 - Article
C2 - 37205712
AN - SCOPUS:85159646820
SN - 0027-8424
VL - 120
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 22
M1 - e2302624120
ER -