Cilia: From Mechanisms to Disease–Part B
- 1st Edition, Volume 176 - April 21, 2023
- Editors: Jose Manuel Bravo-San Pedro, Lorenzo Galluzzi
- Language: English
- Hardback ISBN:9 7 8 - 0 - 4 4 3 - 1 8 5 8 8 - 5
- eBook ISBN:9 7 8 - 0 - 4 4 3 - 1 8 5 8 9 - 2
Cilia: From Mechanisms to Disease Part B, Volume 176 of the Methods in Cell Biology series, presents interesting chapters on topics such as The challenge of dissecting gene… Read more
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Request a sales quoteCilia: From Mechanisms to Disease Part B, Volume 176 of the Methods in Cell Biology series, presents interesting chapters on topics such as The challenge of dissecting gene function in model organisms: tools to characterize genetic mutants and assess transcriptional adaptation in zebrafish, Human LUHMES and NES cells as models for studying primary cilia in neurons, Fixation methods and immunolabeling for cilia proteins in ciliary and extraciliary locations, Single-molecule imaging in the primary cilium, Methods to analyze primary cilia in mouse cardiac lesion model, Methods to study motile ciliated cell types in a zebrafish brain mode, and more.
Other sections cover Clinical and Molecular diagnosis in BBS (Bardet-Biedl syndrome), Modeling ciliopathies in patient-derived primary cells, Analysis of motility and mucociliary function of tracheal epithelial cilia, High-speed Super-resolution SPEED Microscopy to study Live Primary Cilium, Methods for siRNA delivery in retina explants, Methods to study primary cilia and autophagy in the brain, and more.
- Offers a detailed overview of the protocols used to study cilia structure and various aspects of ciliary function
- Provides an approach to the study of some diseases related to ciliary dysfunction, also known as ciliopathies
- Written in an accessible style by renowned experts in the field
Students and entry-level scientists who are for the first time approaching the study of cilia, as well as experienced researchers.
- Cover image
- Title page
- Table of Contents
- Copyright
- Contributors
- Cilia: From Mechanisms to Disease - Part B
- Acknowledgments
- References
- Chapter 1: The challenge of dissecting gene function in model organisms: Tools to characterize genetic mutants and assess transcriptional adaptation in zebrafish
- Abstract
- 1: Introduction
- 2: Methods
- 3: Conclusion
- Acknowledgments
- References
- Chapter 2: Human LUHMES and NES cells as models for studying primary cilia in neurons
- Abstract
- 1: Introduction
- 2: Concluding remarks
- 3: Notes
- Acknowledgments
- References
- Chapter 3: Fixation methods and immunolabeling for cilia proteins in ciliary and extraciliary locations
- Abstract
- 1: Introduction
- 2: Cell culture
- 3: Immunolabeling cilia proteins based on desired area(s) of analysis
- 4: Conclusion
- Acknowledgments
- References
- Chapter 4: Single-molecule imaging in the primary cilium
- Abstract
- 1: Introduction
- 2: Sample preparation
- 3: Super-resolution imaging
- 4: Single-molecule tracking
- 5: Discussion
- 6: Notes
- Acknowledgments
- References
- Chapter 5: Methods for studying primary cilia in heart tissue after ischemia-reperfusion injury
- Abstract
- 1: Introduction
- 2: Myocardial fibrosis induced by ischemia/reperfusion (I/R) injury in mice
- 3: Analysis of primary cilia in heart tissue after I/R
- 4: Isolation of adult mouse fibroblasts and cardiomyocytes for the study of cilia
- 5: Concluding remarks
- 6: Notes
- References
- Chapter 6: Methods to study motile ciliated cell types in the zebrafish brain
- Abstract
- 1: Introduction
- 2: Methods
- 3: Concluding remarks
- 4: Notes
- References
- Chapter 7: Clinical and molecular diagnosis of Bardet-Biedl syndrome (BBS)
- Abstract
- 1: Introduction
- 2: Clinical diagnosis
- 3: Genetic basis and inheritance pattern
- 4: Molecular analysis
- 5: Current management of BBS and future therapies
- Acknowledgments
- References
- Chapter 8: Modeling ciliopathies in patient-derived primary cells
- Abstract
- 1: Introduction
- 2: Patient-derived dermal fibroblast
- 3: Urine derived renal epithelial cells
- 4: Tubuloid preparation from urine derived renal epithelial cells
- 5: Concluding remarks
- 6: Notes
- References
- Chapter 9: Analysis of motility and mucociliary function of tracheal epithelial cilia
- Abstract
- 1: Introduction
- 2: Semi-organ culture of murine trachea
- 3: Analysis of ciliary motility
- 4: Analysis of cilia-driven flow
- 5: Analysis of the motility of ATP-reactivated ciliary axoneme
- 6: Concluding remarks
- 7: Notes
- Acknowledgments
- References
- Chapter 10: High-SPEED super-resolution SPEED microscopy to study primary cilium signaling in vivo
- Abstract
- 1: Introduction
- 2: Concluding Remarks
- Reference
- Chapter 11: Method for siRNA delivery in retina explants
- Abstract
- 1: Introduction
- 2: Reagents and specifications
- 3: Organotypic culture of rat retina explants
- 4: Reverse magnetofection of siRNA in rat retina explants
- 5: Preparation of cryosections from retina explants
- 6: Fluorescent immunohistochemistry (IHC) staining of retina cryosections
- 7: Imaging and analysis of retina cryosections after IHC staining
- 8: Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) of frozen retina sections
- 9: Imaging and analysis of retinal cryosections stained with TUNEL assay
- 10: Western blot of target protein from retina explants
- 11: Concluding remarks
- References
- Chapter 12: Methods to study primary cilia and autophagy in the brain
- Abstract
- 1: Introduction
- 2: Before you begin
- 3: Step-by-step method details
- 4: Expected outcomes
- 5: Quantification and statistical analysis
- 6: Advantages
- 7: Limitations
- 8: Optimization and troubleshooting
- 9: Safety considerations and standards
- Notes
- References
- Chapter 13: Multi-color live-cell fluorescence imaging of primary ciliary membrane assembly and dynamics
- Abstract
- 1: Introduction
- 2: Lentiviral expression plasmid cloning
- 3: Lentivirus production
- 4: Generation of hTERT RPE-1 stable cell line
- 5: Live-cell imaging of ciliary membrane dynamics
- 6: Image analysis
- 7: Concluding remarks
- 8: Notes
- Acknowledgments
- References
- No. of pages: 270
- Language: English
- Edition: 1
- Volume: 176
- Published: April 21, 2023
- Imprint: Academic Press
- Hardback ISBN: 9780443185885
- eBook ISBN: 9780443185892
JB
Jose Manuel Bravo-San Pedro
Jose Manuel Bravo-San Pedro is currently a researcher at the Department of Physiology of the Complutense University of Madrid thanks to a Ramon y Cajal contract grant. He got his Ph.D. in biochemistry, cellular biology and genetics from the University of Extremadura (Caceres, Spain) in 2011, and he did a post-doctoral stage in the laboratory of Prof. Guido Kroemer. His main research interests have always been linked to autophagy, addressing this cellular process associated with neurodegenerative diseases or cancer and recently obesity and specifically related to problems in the correct functioning of the cilium. He is co-inventor of two patents and co-author of 110 publications indexed in PubMed in prestigious international journals, with h-index 45 and 23768 cites (Dec 2022).
Affiliations and expertise
Departamento de Fisiología, Facultad de Medicina, Universidad Complutense de Madrid, Madrid, SpainLG
Lorenzo Galluzzi
Lorenzo Galluzzi is Assistant Professor of Cell Biology in Radiation Oncology at the Department of Radiation Oncology of the Weill Cornell Medical College, Honorary Assistant Professor Adjunct with the Department of Dermatology of the Yale School of Medicine, Honorary Associate Professor with the Faculty of Medicine of the University of Paris, and Faculty Member with the Graduate School of Biomedical Sciences and Biotechnology of the University of Ferrara, the Graduate School of Pharmacological Sciences of the University of Padova, and the Graduate School of Network Oncology and Precision Medicine of the University of Rome “La Sapienza”. Moreover, he is Associate Director of the European Academy for Tumor Immunology and Founding Member of the European Research Institute for Integrated Cellular Pathology.
Galluzzi is best known for major experimental and conceptual contributions to the fields of cell death, autophagy, tumor metabolism and tumor immunology. He has published over 450 articles in international peer-reviewed journals and is the Editor-in-Chief of four journals:
OncoImmunology (which he co-founded in 2011), International Review of Cell and Molecular Biology, Methods in Cell biology, and Molecular and Cellular Oncology (which he co-founded in 2013). Additionally, he serves as Founding Editor for Microbial Cell and Cell Stress, and Associate Editor for Cell Death and Disease, Pharmacological Research and iScience.
Affiliations and expertise
Assistant Professor of Cell Biology in Radiation Oncology, Department of Radiation Oncology, Weill Cornell Medical College, NY, USARead Cilia: From Mechanisms to Disease–Part B on ScienceDirect