Acid-sensing ion channel 3 (ASIC3) cell surface expression is modulated by PSD-95 within lipid rafts

Eshcol, Jayasheel; Harding, Anne Marie S.; Hattori, T.; Costa, Vivian Vasconcelos; Welsh, Michael J.; Benson, Christopher J.

doi:10.1152/ajpcell.00514.2007

Cited by 24 publications

(23 citation statements)

References 53 publications

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“…For electrophysiology experiments, cells were plated in 35-mm tissue culture dishes (40,000 cells/plate) as previously described [11]. After 24 h, CHO-K1 cells were transfected (Transfast, Promega, Madison, WI) with one of the miRNA-carrying plasmids (control, miR847, miR844) and ASIC1a, ASIC3, or ASIC1a and ASIC3 cDNA at varying concentrations (Fig.…”

Section: Methodsmentioning

confidence: 99%

Selective targeting of ASIC3 using artificial miRNAs inhibits primary and secondary hyperalgesia after muscle inflammation

Walder

Gautam

Wilson

et al. 2011

Pain

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Acid-sensing ion channels (ASICs) are activated by acidic pH and may play a significant role in the development of hyperalgesia. Earlier studies show ASIC3 is important for induction of hyperalgesia after muscle insult using ASIC3−/− mice. ASIC3−/− mice lack ASIC3 throughout the body, and the distribution and composition of ASICs could be different from wild-type mice. We therefore tested whether knockdown of ASIC3 in primary afferents innervating muscle of adult wild-type mice prevented development of hyper-algesia to muscle inflammation. We cloned and characterized artificial miRNAs (miR-ASIC3) directed against mouse ASIC3 (mASIC3) to downregulate ASIC3 expression in vitro and in vivo. In CHO-K1 cells transfected with mASIC3 cDNA in culture, the miR-ASIC3 constructs inhibited protein expression of mASIC3 and acidic pH-evoked currents and had no effect on protein expression or acidic pH-evoked currents of ASIC1a. When miR-ASIC3 was used in vivo, delivered into the muscle of mice using a herpes simplex viral vector, both muscle and paw mechanical hyperalgesia were reduced after carrageenan-induced muscle inflammation. ASIC3 mRNA in DRG and protein levels in muscle were decreased in vivo by miRASIC3. In CHO-K1 cells co-transfected with ASIC1a and ASIC3, miR-ASIC3 reduced the amplitude of acidic pH-evoked currents, suggesting an overall inhibition in the surface expression of heteromeric ASIC3-containing channels. Our results show, for the first time, that reducing ASIC3 in vivo in primary afferent fibers innervating muscle prevents the development of inflammatory hyperalgesia in wild-type mice, and thus, may have applications in the treatment of musculoskeletal pain in humans.

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Section: Methodsmentioning

confidence: 99%

Selective targeting of ASIC3 using artificial miRNAs inhibits primary and secondary hyperalgesia after muscle inflammation

Walder

Gautam

Wilson

et al. 2011

Pain

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“…Mouse ASIC1a, ASIC2a, and ASIC3 were previously cloned into the pMT3 plasmid vector, and Chinese hamster ovarian (CHO) cells were transfected as previously described 17. Cells were studied 48–72 hours after transfection.…”

Section: Methodsmentioning

confidence: 99%

“…Whole-cell patch-clamp recordings from sensory neurons and CHO cells were performed as previously described 16, 17. Data are means ± SEM.…”

Section: Methodsmentioning

confidence: 99%

ASIC2a and ASIC3 Heteromultimerize to Form pH-Sensitive Channels in Mouse Cardiac Dorsal Root Ganglia Neurons

Hattori

Chen

Harding

et al. 2009

Circulation Research

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Rationale Acid-Sensing Ion Channels (ASICs) are Na+ channels that are activated by acidic pH. Their expression in cardiac afferents and remarkable sensitivity to small pH changes has made them leading candidates to sense cardiac ischemia. Objective Four genes encode six different ASIC subunits, however it is not yet clear which of the ASIC subunits contribute to the composition of ASICs in cardiac afferents. Methods and Results Here we labeled cardiac afferents using a retrograde tracer dye in mice, which allowed for patch-clamp studies of murine cardiac afferents. We found that a higher percentage of cardiac sensory neurons from the dorsal root ganglia (DRG) respond to acidic pH and generated larger currents compared to those from the nodose ganglia (NG). The ASIC-like current properties of the cardiac DRG neurons from wild-type mice most closely matched the properties of ASIC2a/3 heteromeric channels. This was supported by studies in ASIC null mice: acid-evoked currents from ASIC3 −/− cardiac afferents matched the properties of ASIC2a channels, and currents from ASIC2 −/− cardiac afferents matched the properties of ASIC3 channels. Conclusions We conclude that ASIC2a and -3 are the major ASIC subunits in cardiac DRG neurons, and provide potential molecular targets to attenuate chest pain and deleterious reflexes associated with cardiac disease.

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“…pH 7.4 and pH 5.5 external physiological pH solutions used for the hyaluronic assay and calcium imaging were prepared according to previously published procedures31 (see supplementary material). FLS cultures plated at 100 000 cells/well ( ASIC3 +/+ n=9; ASIC3− / − n=8) were exposed to either pH 7.4 or pH 5.5 for 60 min.…”

Section: Methodsmentioning

confidence: 99%

Acid-sensing ion channel 3 expressed in type B synoviocytes and chondrocytes modulates hyaluronan expression and release

Kolker

Walder

Usachev

et al. 2009

Annals of the Rheumatic Diseases

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Background Rheumatoid arthritis is an inflammatory disease marked by intra-articular decreases in pH, aberrant hyaluronan regulation and destruction of bone and cartilage. Acid-sensing ion channels (ASICs) are the primary acid sensors in the nervous system, particularly in sensory neurons and are important in nociception. ASIC3 was recently discovered in synoviocytes, non-neuronal joint cells critical to the inflammatory process. Objectives To investigate the role of ASIC3 in joint tissue, specifically the relationship between ASIC3 and hyaluronan and the response to decreased pH. Methods Histochemical methods were used to compare morphology, hyaluronan expression and ASIC3 expression in ASIC3+/+ and ASIC3−/− mouse knee joints. Isolated fibroblast-like synoviocytes (FLS) were used to examine hyaluronan release and intracellular calcium in response to decreases in pH. Results In tissue sections from ASIC3+/+ mice, ASIC3 localised to articular cartilage, growth plate, meniscus and type B synoviocytes. In cultured FLS, ASIC3 mRNA and protein was also expressed. In FLS cultures, pH 5.5 increased hyaluronan release in ASIC3+/+ FLS, but not ASIC3−/− FLS. In FLS from ASIC3+/+ mice, approximately 50% of cells (25/53) increased intracellular calcium while only 24% (14/59) showed an increase in ASIC3−/− FLS. Of the cells that responded to pH 5.5, there was significantly less intracellular calcium increases ASIC3−/− FLS compared to ASIC3+/+ FLS. Conclusion ASIC3 may serve as a pH sensor in synoviocytes and be important for modulation of expression of hyaluronan within joint tissue.

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Acid-sensing ion channel 3 (ASIC3) cell surface expression is modulated by PSD-95 within lipid rafts

Cited by 24 publications

References 53 publications

Selective targeting of ASIC3 using artificial miRNAs inhibits primary and secondary hyperalgesia after muscle inflammation

Selective targeting of ASIC3 using artificial miRNAs inhibits primary and secondary hyperalgesia after muscle inflammation

ASIC2a and ASIC3 Heteromultimerize to Form pH-Sensitive Channels in Mouse Cardiac Dorsal Root Ganglia Neurons

Acid-sensing ion channel 3 expressed in type B synoviocytes and chondrocytes modulates hyaluronan expression and release

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