Chondrocytes, the single resident cell type of cartilage, common to both articular and tracheal cartilage, express a number of different voltage- and ligand-gated ion channels, together with ion-selective pumps and exchangers. The expression profile and physiological roles of ion channels in articular chondrocytes are well characterised compared to tracheal chondrocytes, where little is known. Recently, the CaCC, ano1 and ano2, were shown to be essential to tracheal cartilage formation; deletion of ano1 resulted in deformed cartilage rings and premature death in mice [1]. miRNA (miR-30c and miR-125b) have been implicated in the deformation of cartilage rings and the pathogenesis of tracheomalacia, which is characterised by the collapsibility of the trachea [1]. Here, we have characterised the ion channel expression profile of tracheal chondrocytes and compared it to that of articular chondrocytes with a focus on chloride ion channels and their interaction with miRNA. Chondrocytes from rat tracheal and articular cartilage were isolated and the ion channel genes were unbiasedly detected with next generation sequencing (NGS). These datasets was analysed with Ingenuity Pathway Analysis (IPA) and validated with qPCRs. miRNA expression was analysed with qPCR. Patch-clamp electrophysiology was used to characterise the single channel gating of constitutively active ion channel subtypes and pharmacological intervention allowed us to identify VRAC currents as well as CaCC currents. NGS data revealed significant differences between the tracheal and articular chondrocyte ion channel expression profile, with distinct differences in the expression levels of several members of the CaCC, VGCC and VRAC channels, for example, ano1 (A: 494.5, T: 3816.5; p<0.00005; n=8), ano2 (A: 0, T: 18.7; p<0.001; n=8), cacna1g (A: 17060.9, T: 69224.4; p<0.00005; n=8), lrrc8c (A: 16991.1, T: 49684.7; p<0.00005; n=8) and lrrc8d (A: 10636.4, T: 19263.2; p<0.006; n=8). These findings were validated with patch-clamp experiments, where we identified and characterised the single-channel gating of several ion channel subtypes. Using a pharmacological approach we validated the functional expression of VRAC channels with DCPIB (40µM), which blocked VRAC current at ± 100mV (KD: 3.28µM; p<0.02; n=3). The expression of select miRNAs was also analysed, revealing significant differences between the two chondrocyte types, with miR-30c (A: 0.15, T: 0.23; p<0.01; n=6) and miR-141-3p (A: 0.0034, T: 0.0016; p<0.03; n=6) being significantly different. Results are given as mean ± SEM, statistical significance was assessed by one-way ANOVA or Student’s t-tests as appropriate. The ion channel fingerprints between tracheal and articular chondrocytes share several common similarities; yet there were distinct differences observed between the main ion channel families. Future experiments will explore the interplay of the chloride ion channels with miRNA and investigate their impact on chondrocyte function.