Molecular markers expressed in cultured and freshly isolated interstitial cells of Cajal

Epperson, Anne; Hatton, William J.; Callaghan, Brid; Doherty, Philip J.; Walker, Rebecca L.; Sanders, Kenton M.; Ward, Sean M.; Horowitz, Burton

doi:10.1152/ajpcell.2000.279.2.c529

Cited by 159 publications

(164 citation statements)

References 43 publications

(43 reference statements)

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“…This hypothesis also predicts the expression of bipolar ICC with smaller total volume and the lack of ICC network formation within the longitudinal muscle layer where a nonneuronal source of SF supports ICC development (37). Smooth muscle has been shown to preferentially express soluble SF, which is consistent with development of intramuscular ICC due to a brief stimulation of c-kit (4).…”

Section: Discussionmentioning

confidence: 71%

“…Another potential source of SF is smooth muscle. Recent data (4) showed that smooth muscle cells from the murine small intestine express mRNA for the soluble isoform of SF, and smooth muscle cells isolated from the gastric fundus express the membrane-bound isoform of SF mRNA (4). The authors (4) also found that single ICC, selected from the murine gastric fundus or small intestine, express the soluble isoform of SF mRNA.…”

Section: Discussionmentioning

confidence: 99%

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Local presentation of Steel factor increases expression of c-kit immunoreactive interstitial cells of Cajal in culture

Rich¹,

Miller²,

Gibbons

et al. 2003

American Journal of Physiology-Gastrointestinal and Liver Physi

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The binding of Steel factor (SF) to c-kit initiates a signaling pathway essential for development and maintenance of interstitial cells of Cajal (ICC). Soluble and membrane-bound isoforms of SF are expressed in the gastrointestinal tract, but the role for either isoform in supporting ICC development is unknown. The aim of this study was to determine the role of SF in supporting ICC in culture. ICC were cultured from dissociated mouse jejunum and grown with fibroblast cell lines that produced either soluble, membrane-bound or membrane-restricted SF. ICC were identified and counted by c-kit immunoreactivity. The number of c-kit immunoreactive cells was greater in the coculture system compared with cultures grown without SF-producing fibroblasts. All forms of SF-producing fibroblasts increased ICC number in culture but physical separation of the fibroblasts from the c-kit immunoreactive cells, the addition of exogenous SF to the culture medium, or fibroblast-conditioned media did not. These results are consistent with the hypothesis that the membrane-bound form of SF preferentially contributes to expression of c-kit-positive ICC under cell culture conditions.

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Section: Discussionmentioning

confidence: 71%

Section: Discussionmentioning

confidence: 99%

Local presentation of Steel factor increases expression of c-kit immunoreactive interstitial cells of Cajal in culture

Rich¹,

Miller²,

Gibbons

et al. 2003

American Journal of Physiology-Gastrointestinal and Liver Physi

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“…Transcripts of TRPC6 have also been identified in ICC (1). Experiments with BAPTA dialysis in HEK-293 cells stably transfected with TRPC6, however, failed to generate a current similar to that seen in TRPC4␤-transfected cells or in ICC (24).…”

Section: Discussionmentioning

confidence: 99%

TRPC4 currents have properties similar to the pacemaker current in interstitial cells of Cajal

Walker

Koh

Sergeant

et al. 2002

American Journal of Physiology-Cell Physiology

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Interstitial cells of Cajal (ICC) are the pacemaker cells responsible for the generation and propagation of electrical slow waves in phasic muscles of the gastrointestinal (GI) tract. The pacemaker current that initiates each slow wave derives from a calcium-inhibited, voltage-independent, nonselective cation channel. This channel in ICC displays properties similar to that reported for the transient receptor potential (TRP) family of nonselective cation channels, particularly those seen for TRPC3 and TRPC4. We have identified transcripts for TRPC4 in individually isolated ICC and have cloned the two alternatively spliced forms of TRPC4, TRPC4α and TRPC4β, from GI muscles. TRPC4β is missing an 84-amino acid segment from the carboxy terminus. Expression of either form using the whole cell patch-clamp technique led to calcium-inhibited, nonselective cation channels as determined by N-methyl-d-glucamine replacement experiments and BAPTA dialysis. Expression of TRPC4β channels recorded at the whole cell level had characteristics similar to the nonselective cation current in ICC. The single-channel conductance of TRPC4β was determined to be 17.5 pS. Application of calmidazolium to cells expressing TRPC4β led to a significant increase in the inward current of these cells at both the whole cell and single-channel level, and currents were sensitive to block by 10 μM lanthanum, niflumic acid, and DIDS. Comparison of the properties reported for the nonselective cation current in ICC and those identified here for TRPC4β led us to conclude that a TRPC4-like current encodes the plasmalemmal pacemaker current in murine small intestine.

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“…A two-step RT-PCR was performed. The first strand cDNA was synthesized by adding the following components into a nuclease-free microcentrifuge tube in a final 20-l reaction volume: 1 l of oligo(dT) [12][13][14][15][16][17][18] (500 g/ml) (Invitrogen), 2 g of total RNA, 1 l of 10 mM dATP, dGTP, dCTP, and dTTP (dNTP ) mix (Invitrogen), 4 l of 5 ϫ first strand buffer, 2 l of 0.1 M DTT, 1 l of RNase inhibitor, and 1 l of Superscript II RNase H Ϫ reverse transcriptase (Invitrogen). Samples were mixed, incubated at 42°C for 60 min, and inactivated by heating at 70°C for 15 min.…”

Section: Methodsmentioning

confidence: 99%

Superoxide anion is elevated in sympathetic neurons in DOCA-salt hypertension via activation of NADPH oxidase

Dai

Chen²,

Kreulen³

2006

American Journal of Physiology-Heart and Circulatory Physiology

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Ϫ ⅐) production is elevated in sympathetic ganglion neurons and in the vasculature of hypertensive animals; however, it is not known what enzymatic pathway(s) are responsible for O 2 Ϫ ⅐ production. To determine the pathway(s) of O 2 Ϫ ⅐ production in sympathetic neurons, we examined the presence of mRNA of NADPH oxidase subunits in sympathetic ganglionic neurons and differentiated PC-12 cells. The mRNAs for NADPH oxidase subunits p47 phox , p22 phox , gp91 phox , and NOX1 were present in sympathetic neurons and PC-12 cells, whereas the NOX4 homologue was present in sympathetic neurons but not PC-12 cells. Freshly dissociated celiac ganglion neurons from normal rats and PC-12 cells produced O 2 Ϫ ⅐ when treated with the PKC activator PMA; O 2 Ϫ ⅐ production increased by 317% and 254%, respectively. The PMA-evoked increases were reduced by pretreatment with the NADPH oxidase inhibitor apocynin. These findings indicate that NADPH oxidase is the primary source of O 2 Ϫ ⅐ in sympathetic ganglion neurons. When celiac ganglia from hypertensive rats were incubated with apocynin, O 2 Ϫ ⅐ levels were reduced to the same levels as normotensive animals, indicating that NADPH oxidase activity accounted for the elevated O 2 Ϫ ⅐ levels in hypertensive animals. To test this latter finding, we compared NADPH oxidase activity in extracts of prevertebral sympathetic ganglia of DOCA-salt hypertensive rats and shamoperated rats. NADPH oxidase activities were 49.9% and 78.6% higher in sympathetic ganglia of DOCA rats compared with normotensive controls when using ␤-NADH and ␤-NADPH as substrates, respectively. Thus elevated O 2 Ϫ ⅐ levels in hypertension may be a result of the increased activity of NADPH oxidase in postganglionic sympathetic neurons. sympathetic ganglia; phorbol ester; reduced nicotinamide adenine dinucleotide phosphate oxidase; hypertension; deoxycorticosterone acetate VASCULAR ENDOTHELIAL CELLS, smooth muscle cells, and fibroblasts generate reactive oxygen species (ROS), such as O 2 Ϫ ⅐ and H 2 O 2 . ROS play critical roles in both normal and pathophysiological states of vascular cells, including the modulation of redox-sensitive signaling pathways and gene expression, or in the pathophysiology of hypertension and atherosclerosis (16,22,26,36). Several models of hypertension are associated with an increase in vascular O 2 Ϫ ⅐ (41, 42), and this increased O 2 Ϫ ⅐ may quench the endogenous vasodilator nitric oxide (NO) to cause a loss of NO bioactivity in the vessel wall and impair the endothelium-dependent vasorelaxation, resulting in hypertension (26). ROS-mediated endothelial dysfunction in ANG II-infused rats (24) and in DOCA-salt hypertensive rats (42) can be reversed by antioxidant enzyme, endothelial NO synthase (34), or superoxide dismutase (26). ROS can also induce the expression of cardiovascular-related genes, such as those for adhesion molecules and vasoactive substances. For example, the cytokine interleukin 1 activates VCAM-1 gene expression through a mechanism that is repressed ϳ90% by the antioxidant...

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Molecular markers expressed in cultured and freshly isolated interstitial cells of Cajal

Cited by 159 publications

References 43 publications

Local presentation of Steel factor increases expression of c-kit immunoreactive interstitial cells of Cajal in culture

Local presentation of Steel factor increases expression of c-kit immunoreactive interstitial cells of Cajal in culture

TRPC4 currents have properties similar to the pacemaker current in interstitial cells of Cajal

Superoxide anion is elevated in sympathetic neurons in DOCA-salt hypertension via activation of NADPH oxidase

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