Cav3.2 T-type calcium channel is required for the NFAT-dependent Sox9 expression in tracheal cartilage.
Intracellular Ca(2+) transient is crucial in initiating the differentiation of mesenchymal cells into chondrocytes, but whether voltage-gated Ca(2+) channels are involved remains uncertain. Here, we show that the T-type voltage-gated Ca(2+) channel Cav3.2 is essential for tracheal chondrogenesis. Mice lacking this channel (Cav3.2(-/-)) show congenital tracheal stenosis because of incomplete formation of cartilaginous tracheal support. Conversely, Cav3.2 overexpression in ATDC5 cells enhances chondrogenesis, which could be blunted by both blocking T-type Ca(2+) channels and inhibiting calcineurin and suggests that Cav3.2 is responsible for Ca(2+) influx during chondrogenesis. Finally, the expression of sex determination region of Y chromosome (SRY)-related high-mobility group-Box gene 9 (Sox9), one of the earliest markers of committed chondrogenic cells, is reduced in Cav3.2(-/-) tracheas. Mechanistically, Ca(2+) influx via Cav3.2 activates the calcineurin/nuclear factor of the activated T-cell (NFAT) signaling pathway, and a previously unidentified NFAT binding site is identified within the mouse Sox9 promoter using a luciferase reporter assay and gel shift and ChIP studies. Our findings define a previously unidentified mechanism that Ca(2+) influx via the Cav3.2 T-type Ca(2+) channel regulates Sox9 expression through the calcineurin/NFAT signaling pathway during tracheal chondrogenesis.
Institute of Molecular Medicine, National Tsing Hua University, Hsinchu 30013, Taiwan;Institute of Biomedical Sciences, Academia Sinica, Taipei 11529, Taiwan;
Shin-Shiou
SS
Lin
Institute of Biomedical Sciences, Academia Sinica, Taipei 11529, Taiwan;Division of Cardiovascular Medicine, Department of Internal Medicine, Tri-Service General Hospital, National Defense Medical Center, Taipei 11490, Taiwan; and.
Bing-Hsiean
BH
Tzeng
Institute of Molecular Medicine, National Tsing Hua University, Hsinchu 30013, Taiwan;
Kuan-Rong
KR
Lee
Departments of Otolaryngology, Internal Medicine, Pediatrics, and Molecular Physiology and Biophysics.
Richard J H
RJ
Smith
Howard Hughes Medical Institute, andDepartments of Molecular Physiology and Biophysics, Neurology, and Internal Medicine, University of Iowa, Iowa City, IA 52242.
Kevin P
KP
Campbell
Institute of Biomedical Sciences, Academia Sinica, Taipei 11529, Taiwan; ccchen@ibms.sinica.edu.tw.
Chien-Chang
CC
Chen
D000818:Animals; D019703:Calcineurin / Q000378:metabolism; D020747:Calcium Channels, T-Type / Q000235:genetics / Q000502:physiology*; D002356:Cartilage / Q000166:cytology / Q000196:embryology* / Q000502:physiology; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology / Q000502:physiology*; D020219:Chondrogenesis / Q000235:genetics* / Q000502:physiology; D005260:Female; D018507:Gene Expression Regulation, Developmental; D008297:Male; D051379:Mice; D057507:Mice, 129 Strain; D008813:Mice, Inbred ICR; D018345:Mice, Knockout; D050778:NFATC Transcription Factors / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000502:physiology; D055758:SOX9 Transcription Factor / Q000235:genetics* / Q000378:metabolism; D014132:Trachea / Q000166:cytology / Q000196:embryology* / Q000502:physiology
C504883:Cacna1h protein, mouse; D020747:Calcium Channels, T-Type; D050778:NFATC Transcription Factors; D055758:SOX9 Transcription Factor; C527933:Sox9 protein, mouse; D019703:Calcineurin