Abstract
Fluorescent microscope imaging technologies are increasing in their applications and are being used on a wide scale. However methods used to quantify the level of fluorescence intensity are often not utilized—perhaps given the result may be immediately seen, quantification of the data may not seem necessary. However there are a number of reasons given to quantify fluorescent images including the importance of removing potential bias in the data upon observation as well as quantification of large numbers of images gives statistical power to detect subtle changes in experiments. In addition discreet localization of a protein could be detected without selection bias that may not be detectable by eye. Such data will be deemed useful when detecting the levels of HDAC enzymes within cells in order to develop more effective HDAC inhibitor compounds for use against multiple diseased states. Hence, we discuss a methodology devised to analyze fluorescent images using Image J to detect the mean fluorescence intensity of the 11 metal-dependent HDAC enzymes using murine kidney tissue sections as an example.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Chmielewski M, Lindholm B, Stenvinkel P, Ekstrom JT (2011) The role of epigenetics in kidney diseases. Prilozi 32:45–54
Pang M, Kothapally J, Mao H, Tolbert E, Ponnusamy M, Chin YE, Zhuang S (2009) Inhibition of histone deacetylase activity attenuates renal fibroblast activation and interstitial fibrosis in obstructive nephropathy. Am J Physiol Renal Physiol 297:F996–F1005
Bechtel W, McGoohan S, Zeisberg EM et al (2010) Methylation determines fibroblast activation and fibrogenesis in the kidney. Nat Med 16:544–550
Kantharidis P, Wang B, Carew RM, Lan HY (2011) Diabetes complications: the microRNA perspective. Diabetes 60:1832–1837
Mann J, Mann DA (2013) Epigenetic regulation of wound healing and fibrosis. Curr Opin Rheumatol 25:101–107
Perugorria MJ, Wilson CL, Zeybel M, Walsh M, Amin S, Robinson S et al (2012) Histone methyltransferase ASH1 orchestrates fibrogenic gene transcription during myofibroblast transdifferentiation. Hepatology 56:1129–1139
Reddy MA, Natarajan R (2011) Epigenetics in diabetic kidney disease. J Am Soc Nephrol 22:2182–2185
Ververis K, Hiong A, Karagiannis TC, Licciardi PV (2013) Histone deacetylase inhibitors (HDACIs): multitargeted anticancer agents. Biologics 7:47–60
Royce SG, Karagiannis TC (2014) Histone deacetylases and their inhibitors: new implications for asthma and chronic respiratory conditions. Curr Opin Allergy Clin Immunol 14:44–48
Marumo T, Hishikawa K, Yoshikawa M, Hirahashi J, Kawachi S, Fujita T (2010) Histone deacetylase modulates the proinflammatory and -fibrotic changes in tubulointerstitial injury. Am J Physiol Renal Physiol 298:F133–F141
Liu N, He S, Ma L et al (2013) Blocking the class I histone deacetylase ameliorates renal fibrosis and inhibits renal fibroblast activation via modulating TGF-beta and EGFR signaling. PLoS One 8:e54001
Fiji is Just Image J. http://fiji.sc/wiki/index.php/Fiji. Accessed 26 Feb 2015
Gregoretti IV, Lee YM, Goodson HV (2004) Molecular evolution of the histone deacetylase family: functional implications of phylogenetic analysis. J Mol Biol 338(1):17–31
Marks PA, Dokmanovic M (2005) Histone deacetylase inhibitors: discovery and development as anticancer agents. Expert Opin Investig Drugs 14(12):1497–1511
Ververis K, Karagiannis TC (2011) Potential non-oncological applications of histone deacetylase inhibitors. Am J Transl Res 3(5):454–467
Acknowledgements
TDH, CSS, and TK are supported by National Health and Medical Research Council of Australia (NHMRC) Project Grant (APP1078694). TK is an Australian Research Council Future Fellow and CCS a NHMRC Senior Research Fellow (APP1041766).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2016 Springer Science+Business Media New York
About this protocol
Cite this protocol
Ververis, K., Marzully, S., Samuel, C.S., Hewitson, T.D., Karagiannis, T.C. (2016). Qualitative and Quantitative Analysis of Histone Deacetylases in Kidney Tissue Sections. In: Hewitson, T., Smith, E., Holt, S. (eds) Kidney Research. Methods in Molecular Biology, vol 1397. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3353-2_19
Download citation
DOI: https://doi.org/10.1007/978-1-4939-3353-2_19
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3351-8
Online ISBN: 978-1-4939-3353-2
eBook Packages: Springer Protocols