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Analysis of Human Hyaluronan Synthase Gene Transcriptional Regulation and Downstream Hyaluronan Cell Surface Receptor Mobility in Myofibroblast Differentiation

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Glycosaminoglycans

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2303))

Abstract

The ubiquitous extracellular glycosaminoglycan hyaluronan (HA) is a polymer composed of repeated disaccharide units of alternating D-glucuronic acid and D-N-acetylglucosamine residues linked via alternating β-1,4 and β-1,3 glycosidic bonds. Emerging data continue to reveal functions attributable to HA in a variety of physiological and pathological contexts. Defining the mechanisms regulating expression of the human hyaluronan synthase (HAS) genes that encode the corresponding HA-synthesizing HAS enzymes is therefore important in the context of HA biology in health and disease. We describe here methods to analyze transcriptional regulation of the HAS and HAS2-antisense RNA 1 genes. Elucidation of mechanisms of HA interaction with receptors such as the cell surface molecule CD44 is also key to understanding HA function. To this end, we provide protocols for fluorescent recovery after photobleaching analysis of CD44 membrane dynamics in the process of fibroblast to myofibroblast differentiation, a phenotypic transition that is common to the pathology of fibrosis of large organs such as the liver and kidney.

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Acknowledgments

The authors are supported by funding from Kidney Research UK, Medical Research Council UK, Wales Kidney Research Unit, Wellcome Trust, UCB Pharma and the National Natural Science Foundation of China (NSFC) International Young Scholar Research Fellowship (81850410552).

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Authors and Affiliations

  1. Key Laboratory of Bioactive Materials for the Ministry of Education, Rongxiang Xu Center for Regenerative Life Science, College of Life Sciences, Nankai University, Tianjin, China

    Adam C. Midgley

  2. Wales Kidney Research Unit, Division of Infection and Immunity, School of Medicine, College of Biomedical and Life Sciences, Cardiff University, Cardiff, UK

    Timothy Bowen

Authors
  1. Adam C. Midgley
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  2. Timothy Bowen
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Corresponding author

Correspondence to Timothy Bowen .

Editor information

Editors and Affiliations

  1. Departments of Biology, Bioengineering, and Medicinal Chemistry, University of Utah, Salt Lake City, UT, USA

    Kuberan Balagurunathan

  2. Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN, USA

    Hiroshi Nakato

  3. Department of Medicinal Chemistry and Institute for Structural Biology, Drug Discovery and Development, Virginia Commonwealth University, Richmond, VA, USA

    Umesh Desai

  4. Department of Neurobiology & Department of Nutrition & Integrative Physiology, University of Utah, Salt Lake City, UT, USA

    Yukio Saijoh

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Midgley, A.C., Bowen, T. (2022). Analysis of Human Hyaluronan Synthase Gene Transcriptional Regulation and Downstream Hyaluronan Cell Surface Receptor Mobility in Myofibroblast Differentiation. In: Balagurunathan, K., Nakato, H., Desai, U., Saijoh, Y. (eds) Glycosaminoglycans. Methods in Molecular Biology, vol 2303. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1398-6_36

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  • DOI: https://doi.org/10.1007/978-1-0716-1398-6_36

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1397-9

  • Online ISBN: 978-1-0716-1398-6

  • eBook Packages: Springer Protocols

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