Abstract
Erythropoietin (EPO) increases Ca2+ influx in vascular smooth muscle cells and acts both as a direct vasoconstrictor and vascular growth factor (that is, angiogenesis). However, the mechanism by which EPO promotes extracellular Ca2+ entry in contractile cells has not been elucidated. In hematopoietic cells, EPO induces tyrosine kinase (TK)-dependent activation of phospholipase C (PLC)-γ1 and Ca2+ influx via a voltage-independent Ca2+ conductance. In contractile mesangial cells, we have recently characterized a voltage-independent, 1 pS Ca2+ channel that is dependent on both TK and PLC-γ1 activity. Therefore, we examined cultured rat glomerular mesangial cells after timed exposure to recombinant human EPO (20 U/ml). Erythropoietin increased the tyrosine phosphorylation of PLC-γ1, promoted membrane complex formation between PLC-γ1 and the EPO receptor itself, and raised the levels of intracellular inositol 1,4,5-trisphosphate and intracellular Ca2+. Consistent with our previous studies, 1 pS Ca2+ channel activity was extremely low under basal, unstimulated conditions in cell-attached patches, but was dramatically increased when EPO was present in the patch pipette. Tyrosine kinase inhibition with 100 μM genistein or 1 μM PP1 (Src; selective tyrosine kinase inhibitor) prevented all of these EPO-induced responses. We conclude that: (1) EPO-induced stimulation of l pS Ca2+ channels is mediated via a cytosolic Src TK in glomerular mesangial cells. (2) Stimulation of this Ca2+-activated, Ca2+-permeable channel is dependent on the tyrosine phosphorylation/activation of PLC-γ1. (3) This cascade provides a possible mechanism for the vasoconstriction and hypertension observed with clinical EPO use for the treatment of chronic anemias.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 1259-1268 |
| Number of pages | 10 |
| Journal | Kidney International |
| Volume | 53 |
| Issue number | 5 |
| DOIs | |
| State | Published - Jan 1 1998 |
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Keywords
- Anemia
- Hypertension
- Intracellular Ca
- Renal failure
- Tyrosine kinase
ASJC Scopus subject areas
- Nephrology
Cite this
Erythropoietin receptor-operated Ca2+ channels : Activation by phospholipase C-γ1. / Marrero, Mario B.; Venema, Richard C.; Ma, Heping; Ling, Brian N.; Eaton, Douglas C.
In: Kidney International, Vol. 53, No. 5, 01.01.1998, p. 1259-1268.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Erythropoietin receptor-operated Ca2+ channels
T2 - Activation by phospholipase C-γ1
AU - Marrero, Mario B.
AU - Venema, Richard C.
AU - Ma, Heping
AU - Ling, Brian N.
AU - Eaton, Douglas C.
PY - 1998/1/1
Y1 - 1998/1/1
N2 - Erythropoietin (EPO) increases Ca2+ influx in vascular smooth muscle cells and acts both as a direct vasoconstrictor and vascular growth factor (that is, angiogenesis). However, the mechanism by which EPO promotes extracellular Ca2+ entry in contractile cells has not been elucidated. In hematopoietic cells, EPO induces tyrosine kinase (TK)-dependent activation of phospholipase C (PLC)-γ1 and Ca2+ influx via a voltage-independent Ca2+ conductance. In contractile mesangial cells, we have recently characterized a voltage-independent, 1 pS Ca2+ channel that is dependent on both TK and PLC-γ1 activity. Therefore, we examined cultured rat glomerular mesangial cells after timed exposure to recombinant human EPO (20 U/ml). Erythropoietin increased the tyrosine phosphorylation of PLC-γ1, promoted membrane complex formation between PLC-γ1 and the EPO receptor itself, and raised the levels of intracellular inositol 1,4,5-trisphosphate and intracellular Ca2+. Consistent with our previous studies, 1 pS Ca2+ channel activity was extremely low under basal, unstimulated conditions in cell-attached patches, but was dramatically increased when EPO was present in the patch pipette. Tyrosine kinase inhibition with 100 μM genistein or 1 μM PP1 (Src; selective tyrosine kinase inhibitor) prevented all of these EPO-induced responses. We conclude that: (1) EPO-induced stimulation of l pS Ca2+ channels is mediated via a cytosolic Src TK in glomerular mesangial cells. (2) Stimulation of this Ca2+-activated, Ca2+-permeable channel is dependent on the tyrosine phosphorylation/activation of PLC-γ1. (3) This cascade provides a possible mechanism for the vasoconstriction and hypertension observed with clinical EPO use for the treatment of chronic anemias.
AB - Erythropoietin (EPO) increases Ca2+ influx in vascular smooth muscle cells and acts both as a direct vasoconstrictor and vascular growth factor (that is, angiogenesis). However, the mechanism by which EPO promotes extracellular Ca2+ entry in contractile cells has not been elucidated. In hematopoietic cells, EPO induces tyrosine kinase (TK)-dependent activation of phospholipase C (PLC)-γ1 and Ca2+ influx via a voltage-independent Ca2+ conductance. In contractile mesangial cells, we have recently characterized a voltage-independent, 1 pS Ca2+ channel that is dependent on both TK and PLC-γ1 activity. Therefore, we examined cultured rat glomerular mesangial cells after timed exposure to recombinant human EPO (20 U/ml). Erythropoietin increased the tyrosine phosphorylation of PLC-γ1, promoted membrane complex formation between PLC-γ1 and the EPO receptor itself, and raised the levels of intracellular inositol 1,4,5-trisphosphate and intracellular Ca2+. Consistent with our previous studies, 1 pS Ca2+ channel activity was extremely low under basal, unstimulated conditions in cell-attached patches, but was dramatically increased when EPO was present in the patch pipette. Tyrosine kinase inhibition with 100 μM genistein or 1 μM PP1 (Src; selective tyrosine kinase inhibitor) prevented all of these EPO-induced responses. We conclude that: (1) EPO-induced stimulation of l pS Ca2+ channels is mediated via a cytosolic Src TK in glomerular mesangial cells. (2) Stimulation of this Ca2+-activated, Ca2+-permeable channel is dependent on the tyrosine phosphorylation/activation of PLC-γ1. (3) This cascade provides a possible mechanism for the vasoconstriction and hypertension observed with clinical EPO use for the treatment of chronic anemias.
KW - Anemia
KW - Hypertension
KW - Intracellular Ca
KW - Renal failure
KW - Tyrosine kinase
UR - http://www.scopus.com/inward/record.url?scp=0031870588&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0031870588&partnerID=8YFLogxK
U2 - 10.1046/j.1523-1755.1998.00887.x
DO - 10.1046/j.1523-1755.1998.00887.x
M3 - Article
C2 - 9573541
AN - SCOPUS:0031870588
VL - 53
SP - 1259
EP - 1268
JO - Kidney International
JF - Kidney International
SN - 0085-2538
IS - 5
ER -