Hyaluronan synthesis by developing cortical neurons in vitro

Hyaluronan is a linear glycosaminoglycan that forms the backbone of perineuronal nets around neurons in the cerebral cortex. However, it remains controversial whether neurons are capable of independent hyaluronan synthesis. Herein, we examined the expression of hyaluronan and hyaluronan synthases (H...

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Published in:Scientific reports 2017-03, Vol.7 (1), p.44135-44135, Article 44135
Main Authors: Fowke, Tania M., Karunasinghe, Rashika N., Bai, Ji-Zhong, Jordan, Shawn, Gunn, Alistair J., Dean, Justin M.
Format: Article
Language:English
Subjects:
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Axonogenesis
Cell culture
Cerebral cortex
Dendrites
Filopodia
Growth cones
Humanities and Social Sciences
Hyaluronic acid
Immunocytochemistry
Lamellipodia
mRNA
multidisciplinary
Nets
Neuronal-glial interactions
Neurons
Perineuronal nets
Rodents
Science
Synapses
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Summary:Hyaluronan is a linear glycosaminoglycan that forms the backbone of perineuronal nets around neurons in the cerebral cortex. However, it remains controversial whether neurons are capable of independent hyaluronan synthesis. Herein, we examined the expression of hyaluronan and hyaluronan synthases (HASs) throughout cortical neuron development in vitro . Enriched cultures of cortical neurons were established from E16 rats. Neurons were collected at days in vitro (DIV) 0 (4 h), 1, 3, 7, 14, and 21 for qPCR or immunocytochemistry. In the relative absence of glia, neurons exhibited HAS1–3 mRNA at all time-points. By immunocytochemistry, puncta of HAS2–3 protein and hyaluronan were located on neuronal cell bodies, neurites, and lamellipodia/growth cones from as early as 4 h in culture. As neurons matured, hyaluronan was also detected on dendrites, filopodia, and axons, and around synapses. Percentages of hyaluronan-positive neurons increased with culture time to ~93% by DIV21, while only half of neurons at DIV21 expressed the perineuronal net marker Wisteria floribunda agglutinin. These data clearly demonstrate that neurons in vitro can independently synthesise hyaluronan throughout all maturational stages, and that hyaluronan production is not limited to neurons expressing perineuronal nets. The specific structural localisation of hyaluronan suggests potential roles in neuronal development and function.
ISSN:2045-2322
2045-2322
DOI:10.1038/srep44135