Clcn6 mRNA and ClC-6 protein expression in mouse central nervous system

Abstract

To investigate the expression patterns of Clcn6 mRNA and ClC-6 protein in postnatal mouse of distinct region of central nervous system (CNS) and developmental stages. Distinct CNS regions, including cerebral cortex, hippocampus, cerebellum, brainstem, olfactory bulb, hypothalamus, corpus striatum, thalamus and spinal cord, were dissected from C57BL/6 wildtype mice at different developmental stages, including P0, P7, P14, P30, and P60. Total RNA and protein were extracted. Quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) and western blot analysis were performed to evaluate Clcn6 mRNA and ClC-6 protein expression levels. In addition, publicly available datasets, including the Human Protein Atlas (HPA) mouse brain RNA-seq dataset and the Allen Mouse Brain ISH dataset were analyzed for comparative assessment of regional Clcn6 mRNA expression patterns. Clcn6 mRNA and ClC-6 protein were detected in all examined brain regions and spinal cord throughout postnatal development and into adulthood. Relatively higher Clcn6 mRNA expression levels were observed at early postnatal stages in the cerebral cortex, hippocampus, cerebellum, hypothalamus, and olfactory bulb. ClC-6 protein expression remained relatively stable in the cerebral cortex, hippocampus, cerebellum, hypothalamus, and olfactory bulb during postnatal development. In adult mice, relatively higher Clcn6 mRNA expression was observed in the hypothalamus, whereas relatively lower expression was observed in the corpus striatum. ClC-6 protein levels were relatively higher in the cerebellum and hypothalamus and lower in the corpus striatum and brainstem. Collation and analysis of two public databases further supported regional differences in Clcn6 mRNA expression. Clcn6 is widely expressed throughout the mouse CNS during postnatal development and exhibits region- and developmental stage-related expression patterns. These findings provide a descriptive expression profile of Clcn6 in the mouse CNS and may contribute to future studies investigating the physiological role of ClC-6 in neural development and function.