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Tissue distribution and subcellular localization of hyaluronan synthase isoenzymes

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Abstract

Hyaluronan synthases (HAS) are unique plasma membrane glycosyltransferases secreting this glycosaminoglycan directly to the extracellular space. The three HAS isoenzymes (HAS1, HAS2, and HAS3) expressed in mammalian cells differ in their enzymatic properties and regulation by external stimuli, but clearly distinct functions have not been established. To overview the expression of different HAS isoenzymes during embryonic development and their subcellular localization, we immunostained mouse embryonic samples and cultured cells with HAS antibodies, correlating their distribution to hyaluronan staining. Their subcellular localization was further studied by GFP–HAS fusion proteins. Intense hyaluronan staining was observed throughout the development in the tissues of mesodermal origin, like heart and cartilages, but also for example during the maturation of kidneys and stratified epithelia. In general, staining for one or several HASs correlated with hyaluronan staining. The staining of HAS2 was most widespread, both spatially and temporally, correlating with hyaluronan staining especially in early mesenchymal tissues and heart. While epithelial cells were mostly negative for HASs, stratified epithelia became HAS positive during differentiation. All HAS isoenzymes showed cytoplasmic immunoreactivity, both in tissue sections and cultured cells, while plasma membrane staining was also detected, often in cellular extensions. HAS1 had brightest signal in Golgi, HAS3 in Golgi and microvillous protrusions, whereas most of the endogenous HAS2 immunoreactivity was localized in the ER. This differential pattern was also observed with transfected GFP–HASs. The large proportion of intracellular HASs suggests that HAS forms a reserve that is transported to the plasma membrane for rapid activation of hyaluronan synthesis.

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Acknowledgments

Special thanks are due to Eija Kettunen, Kari Kotikumpu, Eija Rahunen, and Arja Venäläinen for expert technical assistance. This work was supported by the Academy of Finland, Grant #40807 and #54062 (M.T.), and by the grants from The North Savo Cultural Foundation (K.R.), Kuopio University Foundation (K.R.), The North Savo Cancer Foundation (K.R), Paavo Koistinen Foundation (K.R. and K.T.), The Finnish Cancer Foundation (R.T.), Sigrid Juselius Foundation (RT and MT), The Spearhead Funds of the University of Eastern Finland (Cancer Center of Eastern Finland), and the EVO funds of Kuopio University Hospital (M.T.).

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  1. Institute of Biomedicine, Anatomy, University of Eastern Finland, P.O.B. 1627, 70211, Kuopio, Finland

    Kari Törrönen, Kaisa Nikunen, Riikka Kärnä, Markku Tammi, Raija Tammi & Kirsi Rilla

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  1. Kari Törrönen
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Correspondence to Kirsi Rilla.

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Törrönen, K., Nikunen, K., Kärnä, R. et al. Tissue distribution and subcellular localization of hyaluronan synthase isoenzymes. Histochem Cell Biol 141, 17–31 (2014). https://doi.org/10.1007/s00418-013-1143-4

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