Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Identification and Characterization of microRNAs during Retinoic Acid-Induced Regeneration of a Molluscan Central Nervous System

Version 1 : Received: 6 September 2018 / Approved: 7 September 2018 / Online: 7 September 2018 (03:13:31 CEST)

How to cite: Walker, S.E.; Spencer, G.E.; Necakov, A.; Carlone, R.L. Identification and Characterization of microRNAs during Retinoic Acid-Induced Regeneration of a Molluscan Central Nervous System. Preprints 2018, 2018090123. https://doi.org/10.20944/preprints201809.0123.v1 Walker, S.E.; Spencer, G.E.; Necakov, A.; Carlone, R.L. Identification and Characterization of microRNAs during Retinoic Acid-Induced Regeneration of a Molluscan Central Nervous System. Preprints 2018, 2018090123. https://doi.org/10.20944/preprints201809.0123.v1

Abstract

Retinoic acid (RA) is the biologically active metabolite of vitamin A and has become a well-established factor that induces neurite outgrowth and regeneration in both vertebrates and invertebrates. However, the underlying regulatory mechanisms that may mediate RA-induced neurite sprouting remain unclear. In the past decade, microRNAs have emerged as important regulators of nervous system development and regeneration, and have been shown to contribute to processes such as neurite sprouting. However, few studies have demonstrated the role of miRNAs in RA-induced neurite sprouting. By miRNA-Sequencing analysis, we identify 482 miRNAs in the regenerating CNS of the mollusc Lymnaea stagnalis, 219 of which represent potentially novel miRNAs. Of the remaining conserved miRNAs, 38 show a statistically significant up or downregulation in regenerating CNS as a result of RA treatment. We further characterized the expression of one neuronally-enriched miRNA upregulated by RA, miR-124. We demonstrate for the first time that miR-124 is expressed within the cell bodies and neurites of regenerating motorneurons. Moreover, we identify miR-124 expression within the growth cones of cultured ciliary motorneurons (Pedal A), whereas expression in the growth cones of another class of respiratory motorneurons (RPA) was absent in vitro. These findings support our hypothesis that miRNAs are important regulators of retinoic acid-induced neuronal outgrowth and regeneration in regeneration-competent species.

Keywords

retinoic acid; microRNA; RNA sequencing; neuronal regeneration; growth cone; Lymnaea

Subject

Biology and Life Sciences, Anatomy and Physiology

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