Transcriptomic Profiling of Ca2+ Transport Systems During the Formation of the Cerebral Cortex in Mice

Abstract

Cytosolic calcium (Ca²⁺) transients control key neural processes, including neurogenesis, migration, the polarization and growth of neurons, and the establishment and maintenance of synaptic connections. They are thus involved in the development and formation of the neural system. In this study, a publicly available whole transcriptome sequencing (RNA-Seq) dataset was used to examine the expression of genes coding for putative plasma membrane and organellar Ca²⁺-transporting proteins (channels, pumps, exchangers, and transporters) during the formation of the cerebral cortex in mice. Four ages were considered: embryonic days 11 (E11), 13 (E13), and 17 (E17), and post-natal day 1 (PN1). This transcriptomic profiling was also combined with live-cell Ca²⁺ imaging recordings to assess the presence of functional Ca²⁺ transport systems in E13 neurons. The most important Ca²⁺ routes of the cortical wall at the onset of corticogenesis (E11-E13) were TACAN, GluK5, nAChR β2, Cav3.1, Orai3, transient receptor potential cation channel subfamily M member 7 (TRPM7) non-mitochondrial Na⁺/Ca²⁺ exchanger 2 (NCX2), and the connexins CX43/CX45/CX37. Hence, transient receptor potential cation channel mucolipin subfamily member 1 (TRPML1), transmembrane protein 165 (TMEM165), and Ca²⁺ "leak" channels are prominent intracellular Ca²⁺ pathways. The Ca²⁺ pumps sarco/endoplasmic reticulum Ca²⁺ ATPase 2 (SERCA2) and plasma membrane Ca²⁺ ATPase 1 (PMCA1) control the resting basal Ca²⁺ levels. At the end of neurogenesis (E17 and onward), a more numerous and diverse population of Ca²⁺ uptake systems was observed. In addition to the actors listed above, prominent Ca²⁺-conducting systems of the cortical wall emerged, including acid-sensing ion channel 1 (ASIC1), Orai2, P2X2, and GluN1. Altogether, this study provides a detailed view of the pattern of expression of the main actors participating in the import, export, and release of Ca²⁺. This work can serve as a framework for further functional and mechanistic studies on Ca²⁺ signaling during cerebral cortex formation.

Keywords: Na/Ca exchanger; calcium channels; calcium pumps; calcium stores; cerebral cortex; endoplasmic reticulum; neurogenesis.