Channel properties of the splicing isoforms of the olfactory calcium-activated chloride channel Anoctamin 2

Abstract

Figure 1. Characterization of Ano2 mRNA variants present in mouse olfactory epithelium. (A) Diagram summarizing the 5 Ano2 exon splicing structure. The green sections indicate the most 5 AUG translation start codons, and the subsequent black bars indicate predicted protein-coding sequence. The variants containing exons 1a and 2 are less abundant in the olfactory epithelium than the variants containing the newly discovered exon 1b (red), as determined in B and C. The five forward (F1-F5) and one reverse (R1) PCR primer-binding sites are indicated as arrows. Given two alternative 5 ends and alternative splicing of exon 4, these mRNA variants may encode up to four ANO2 isoforms, named on the right. (B and C) Ethidium bromide-stained agarose gels showing RT-PCR products with primers specific for each 5 variant of Ano2. Primer F1 is universal for both variants, F2 is specific for exon 1b, F3 and F4 are specific for exon 2, and F5 is specific for exon 1a. The reverse primer (R1) was the same for all PCR reactions. The expected PCR product sizes are 267, 409, 350, 511, and 830 bp, respectively. Quantification of the bands by densitometry is shown in Fig. S1 C.

Honokiol inhibits proliferation of colorectal cancer cells by targeting anoctamin 1/TMEM16A Ca2+‐activated Cl− channels

“…The cDNAs for the Ano1(ac) isoform (NCBI accession number: NP_848757) containing alternative splicing exons 1 and 13 and lacking exons 6b and 15 (Caputo et al, 2008) or the Ano2 isoform (NCBI accession number: XP_006506182) containing alternative splicing exons 4 and 13 and lacking exons 1 and 2 (Ponissery Saidu et al, 2013) were cloned into the pEGFP‐N1 vectors. PCR‐based site‐directed mutagenesis was performed to make mutations using Phanta Max Super‐Fidelity DNA Polymerase (Vazyme, China).…”

Elevated Cytosolic Cl−Concentrations in Dendritic Knobs of Mouse Vomeronasal Sensory Neurons

“…Similar to the Ca 2+ -dependent Cl − conductance known to amplify transduction currents in canonical olfactory neurons (Pifferi et al 2009;Stephan et al 2009;Sagheddu et al 2010;Billig et al 2011;Dauner et al 2012;Ponissery-Saidu et al 2013;Henkel et al 2014), a considerable amount of stimulus-evoked VSN activity seems to be carried by a Ca…”