Immediate-Early Genes Detection in the CNS of Terrestrial Snail
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ORIGINAL RESEARCH
Immediate‑Early Genes Detection in the CNS of Terrestrial Snail Victor N. Ierusalimsky1 · Matvey V. Roshchin1 · Pavel M. Balaban1 Received: 26 November 2019 / Accepted: 2 March 2020 © Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract In the present work, using in situ hybridization, we studied the expression patterns of three molluscan homologs of vertebrate immediate-early genes C/EBP, c-Fos, and c-Jun in the central nervous system (CNS) of terrestrial gastropod snail Helix. The molluscan C/EBP gene was described in literature, while c-Fos and c-Jun were studied in terrestrial snails for the first time. Localization of the expression was traced in normal conditions, and in preparations physiologically activated using stimulation of suboesophageal ganglia nerves. No expression was detected constitutively. In stimulated preparations, all three genes had individual expression patterns in Helix CNS, and the level of expression was stimulus-dependent. The number of cells expressing the gene of interest was different from the number of cells projecting to the stimulated nerve, and thus activated retrogradely. This difference depended on the ganglia studied. At the subcellular level, the labeled RNA was observed as dots (probably small clusters of RNA molecules) and shapeless mass of RNA, often seen as a circle at the internal border of the cell nuclei. The data provide a basis for further study of behavioral role of these putative immediateearly genes in snail behavior and learning. Keywords c-Fos · c-Jun · C/EBP · Gastropods · Terrestrial snail · Neurons
Introduction Immediate-early genes (IEGs) constitute the population of effector genes which are rapidly upregulated by the sensory stimuli, and most of them are involved in the transcriptional regulation of other upregulated genes expressed with a delay (Gonzales et al. 2019; Kim et al. 2018; Pérez-Cadahía et al. 2011; Clayton 2000; Clayton et al. 2019). It is typical that, expression of the IEGs (i) is mostly absent in quiescent state and is induced by stimuli of many modalities, (ii) occurs within minutes, and (iii) takes place in the absence of de novo protein synthesis. A lot of studies devoted to the key role of the IEGs in development, learning and memory were made in vertebrates, and numerous sets of IEGs were identified (for rev., see Korzus 2003; Kaczmarek 2000; Okuno 2011; Clayton et al. 2019). In invertebrates, mollusks in particular, only several transcriptional factors homologous to the vertebrate IEGs were identified. Among them, CREB * Victor N. Ierusalimsky [email protected] 1
Institute of Higher Nervous Activity and Neurophysiology, Russian Academy of Sciences, 5A Butlerova str., Moscow, Russia 117485
was well studied due to its important role in the organization of behavior. It is known to increase transcription of other IEGs (Clayton et al. 2019). CREB1 was identified in Aplysia (Lewin and Walters 1999; Liu et al. 2008) and Lymnaea (Sadamoto et al; 2004a, b; 2010). But till now the role of m
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