Abstract
Kidneys are highly aerobic organs and their function is tightly coupled to mitochondrial energy production. Renal tubular cells, particularly the proximal tubule (PT), require an abundance of mitochondria to provide sufficient energy for regulating fluid and electrolyte balance. Meanwhile, mitochondrial defects are implicated in a range of different kidney diseases. Multiphoton microscopy (MP) is a powerful tool that allows detailed study of mitochondrial morphology, dynamics, and function in kidney tissue. Here, we describe how MP can be used to image mitochondria in kidney tubular cells, either ex vivo in tissue slices or in vivo in living rodents, using both endogenous and exogenous fluorescent molecules. Moreover, changes in mitochondrial signals can be followed in real time in response to different insults or stimuli, in parallel with other important readouts of kidney tubular function, such as solute uptake and trafficking.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Bagnasco S, Good D, Balaban R, Burg M (1985) Lactate production in isolated segments of the rat nephron. Am J Physiol 248:F522–F526
Fujita K (2007) Two-photon laser scanning fluorescence microscopy. Tanpakushitsu Kakusan Koso 52:1778–1779
Hall AM, Unwin RJ, Parker N, Duchen MR (2009) Multiphoton imaging reveals differences in mitochondrial function between nephron segments. J Am Soc Nephrol 20:1293–1302
Weigert R, Sramkova M, Parente L et al (2010) Intravital microscopy: a novel tool to study cell biology in living animals. Histochem Cell Biol 133:481–491
Molitoris BA, Sandoval RM (2005) Intravital multiphoton microscopy of dynamic renal processes. Am J Physiol 288:1084–1089
Peti-Peterdi J, Toma I, Sipos A, Vargas SL (2009) Multiphoton imaging of renal regulatory mechanisms. Physiology 24:88–96
Hall AM, Crawford C, Unwin RJ et al (2011) Multiphoton imaging of the functioning kidney. J Am Soc Nephrol 22:1297–1304
Bugarski M, Martins JR, Haenni D, Hall AM (2018) Multiphoton imaging reveals axial differences in metabolic autofluorescence signals along the kidney proximal tubule. Am J Physiol Renal Physiol 315:F1613–F1625
Scaduto RC, Grotyohann LW (1999) Measurement of mitochondrial membrane potential using fluorescent rhodamine derivatives. Biophys J 76:469–477
Bassnett S, Reinisch L, Beebe DC (1990) Intracellular pH measurement using single excitation-dual emission fluorescence ratios. Am J Physiol 258:C171–C178
Chance B, Schoener B, Oshino R et al (1979) Oxidation-reduction ratio studies of mitochondria in freeze-trapped samples. NADH and flavoprotein fluorescence signals. J Biol Chem 254:4764–4771
Mayevsky A, Chance B (2007) Oxidation-reduction states of NADH in vivo: from animals to clinical use. Mitochondrion 7:330–339
Dunn KW, Sutton TA, Sandoval RM (2007) Live-animal imaging of renal function by multiphoton microscopy. Curr Protoc Cytom 41:12.9.1–12.9.18
Gekle M, Mildenberger S, Freudinger R, Silbernagl S (1995) Endosomal alkalinization reduces J(max) and K(m) of albumin receptor- mediated endocytosis in OK cells. Am J Physiol 268:F899–F906
Wang E, Sandoval RM, Campos SB, Molitoris BA (2010) Rapid diagnosis and quantification of acute kidney injury using fluorescent ratio-metric determination of glomerular filtration rate in the rat. Am J Physiol Renal Physiol 299:1048–1055
Perry SW, Norman JP, Barbieri J et al (2011) Mitochondrial membrane potential probes and the proton gradient: a practical usage guide. BioTechniques 50:98–115
Owen S (1992) Comparison of Spectrum-shifting intracellular pH. Anal Biochem 204:65–71
Acknowledgments
This work was supported by The Swiss National Centre for Competence in Research (NCCR) Kidney Control of Homeostasis and by a project grant from the Swiss National Science Foundation. The authors also acknowledge support from The Zurich Centre for Microscopy and Image Analysis and The Zurich Centre for Integrative Human Physiology.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2021 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Bugarski, M., Ghazi, S., Hall, A.M. (2021). Live Imaging of Mitochondria in Kidney Tissue. In: Weissig, V., Edeas, M. (eds) Mitochondrial Medicine . Methods in Molecular Biology, vol 2275. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1262-0_25
Download citation
DOI: https://doi.org/10.1007/978-1-0716-1262-0_25
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-1261-3
Online ISBN: 978-1-0716-1262-0
eBook Packages: Springer Protocols