Tubular and cellular localization of the cardiac L-type calcium channel in rat kidney

Pei Lin Zhao, Xi Tao Wang, Xue Mei Zhang, Valeriu Cebotaru, Liudmila Cebotaru, Gang Guo, Marcelo Morales, Sandra E. Guggino

Research output: Contribution to journalArticle

Abstract

Background. The mRNAs of several types of calcium channels have been identified in intact rat kidney, and L-type calcium channels cause changes in intracellular calcium in primary cultures of distal tubule cells. The aim of this study was to evaluate the tubular and cellular distribution of the α1C subunit of the L-type calcium channel in intact kidney. Methods. RT-PCR and Northern blot analysis were used to assess the regional abundance of the mRNA of this channel. Immunocytochemistry combined with confocal microscopy and surface biotinylation were applied to determine the tubular and cellular localization of the protein. Results. Northern blot and RT-PCR analysis indicated that the mRNA of the α1C subunit of the cardiac L-type calcium channel was present in whole rat kidney, kidney tubules and kidney cell lines. Western blot of lysates from whole kidney, kidney tubules or cell lines revealed bands of ∼190 kD for the α1C subunit and ∼60 kD for the β3 subunit. Confocal immunohistochemistry indicated that the α1C subunit of this channel was co-expressed in cells of the distal tubule that express calbindin-D28K, but not in intercalated cells. The α1C subunit was also highly expressed in both outer and inner medullary collecting ducts. Serial confocal microscopic images or surface biotinylation experiments determined that the channel was predominantly on the basolateral membrane but had some distribution on the apical membrane. Conclusions. The distribution and cellular localization of the α1C subunit of cardiac L-type calcium channel suggest it is probably involved in intracellular and membrane calcium signaling.

Original languageEnglish (US)
Pages (from-to)1393-1406
Number of pages14
JournalKidney International
Volume61
Issue number4
DOIs
StatePublished - 2002

Fingerprint

L-Type Calcium Channels
Kidney
Kidney Tubules
Biotinylation
Northern Blotting
Messenger RNA
Immunohistochemistry
Calbindin 1
Cell Line
Polymerase Chain Reaction
Intracellular Membranes
Calcium Signaling
Membranes
Calcium Channels
Confocal Microscopy
Western Blotting
Calcium
Proteins

Keywords

  • Calcium signaling
  • Cortical collecting duct
  • Distal tubule
  • Intracellular signaling
  • Medullary collecting duct

ASJC Scopus subject areas

  • Nephrology

Cite this

Zhao, P. L., Wang, X. T., Zhang, X. M., Cebotaru, V., Cebotaru, L., Guo, G., ... Guggino, S. E. (2002). Tubular and cellular localization of the cardiac L-type calcium channel in rat kidney. Kidney International, 61(4), 1393-1406. https://doi.org/10.1046/j.1523-1755.2002.00267.x

Tubular and cellular localization of the cardiac L-type calcium channel in rat kidney. / Zhao, Pei Lin; Wang, Xi Tao; Zhang, Xue Mei; Cebotaru, Valeriu; Cebotaru, Liudmila; Guo, Gang; Morales, Marcelo; Guggino, Sandra E.

In: Kidney International, Vol. 61, No. 4, 2002, p. 1393-1406.

Research output: Contribution to journalArticle

Zhao, PL, Wang, XT, Zhang, XM, Cebotaru, V, Cebotaru, L, Guo, G, Morales, M & Guggino, SE 2002, 'Tubular and cellular localization of the cardiac L-type calcium channel in rat kidney', Kidney International, vol. 61, no. 4, pp. 1393-1406. https://doi.org/10.1046/j.1523-1755.2002.00267.x
Zhao, Pei Lin ; Wang, Xi Tao ; Zhang, Xue Mei ; Cebotaru, Valeriu ; Cebotaru, Liudmila ; Guo, Gang ; Morales, Marcelo ; Guggino, Sandra E. / Tubular and cellular localization of the cardiac L-type calcium channel in rat kidney. In: Kidney International. 2002 ; Vol. 61, No. 4. pp. 1393-1406.
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AU - Zhao, Pei Lin

AU - Wang, Xi Tao

AU - Zhang, Xue Mei

AU - Cebotaru, Valeriu

AU - Cebotaru, Liudmila

AU - Guo, Gang

AU - Morales, Marcelo

AU - Guggino, Sandra E.

PY - 2002

Y1 - 2002

N2 - Background. The mRNAs of several types of calcium channels have been identified in intact rat kidney, and L-type calcium channels cause changes in intracellular calcium in primary cultures of distal tubule cells. The aim of this study was to evaluate the tubular and cellular distribution of the α1C subunit of the L-type calcium channel in intact kidney. Methods. RT-PCR and Northern blot analysis were used to assess the regional abundance of the mRNA of this channel. Immunocytochemistry combined with confocal microscopy and surface biotinylation were applied to determine the tubular and cellular localization of the protein. Results. Northern blot and RT-PCR analysis indicated that the mRNA of the α1C subunit of the cardiac L-type calcium channel was present in whole rat kidney, kidney tubules and kidney cell lines. Western blot of lysates from whole kidney, kidney tubules or cell lines revealed bands of ∼190 kD for the α1C subunit and ∼60 kD for the β3 subunit. Confocal immunohistochemistry indicated that the α1C subunit of this channel was co-expressed in cells of the distal tubule that express calbindin-D28K, but not in intercalated cells. The α1C subunit was also highly expressed in both outer and inner medullary collecting ducts. Serial confocal microscopic images or surface biotinylation experiments determined that the channel was predominantly on the basolateral membrane but had some distribution on the apical membrane. Conclusions. The distribution and cellular localization of the α1C subunit of cardiac L-type calcium channel suggest it is probably involved in intracellular and membrane calcium signaling.

AB - Background. The mRNAs of several types of calcium channels have been identified in intact rat kidney, and L-type calcium channels cause changes in intracellular calcium in primary cultures of distal tubule cells. The aim of this study was to evaluate the tubular and cellular distribution of the α1C subunit of the L-type calcium channel in intact kidney. Methods. RT-PCR and Northern blot analysis were used to assess the regional abundance of the mRNA of this channel. Immunocytochemistry combined with confocal microscopy and surface biotinylation were applied to determine the tubular and cellular localization of the protein. Results. Northern blot and RT-PCR analysis indicated that the mRNA of the α1C subunit of the cardiac L-type calcium channel was present in whole rat kidney, kidney tubules and kidney cell lines. Western blot of lysates from whole kidney, kidney tubules or cell lines revealed bands of ∼190 kD for the α1C subunit and ∼60 kD for the β3 subunit. Confocal immunohistochemistry indicated that the α1C subunit of this channel was co-expressed in cells of the distal tubule that express calbindin-D28K, but not in intercalated cells. The α1C subunit was also highly expressed in both outer and inner medullary collecting ducts. Serial confocal microscopic images or surface biotinylation experiments determined that the channel was predominantly on the basolateral membrane but had some distribution on the apical membrane. Conclusions. The distribution and cellular localization of the α1C subunit of cardiac L-type calcium channel suggest it is probably involved in intracellular and membrane calcium signaling.

KW - Calcium signaling

KW - Cortical collecting duct

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