Selectively enhanced cellular signaling by Gi proteins in essential hypertension. G alpha i2, G alpha i3, G beta 1, and G beta 2 are not mutated.
Recent studies have shown an enhanced signaling capacity of receptors coupled to pertussis toxin (PTX)-sensitive guanine nucleotide-binding proteins (G proteins) in immortalized B lymphoblasts from patients with essential hypertension. In the present study, we analyzed (1) whether such alterations would also be expressed in nontransformed cells of these individuals and (2) whether other G protein-mediated signaling pathways were also altered. Therefore, we established primary cultures of skin fibroblasts from previously characterized normotensive and hypertensive individuals (NT and HT cells, respectively). [Ca2+]i rises induced by lyso-phosphatidic acid (LPA), thrombin, and sphingosine-1-phosphate as well as the formation of inositol 1,4,5-trisphosphate and [3H]thymidine incorporation evoked by LPA were PTX sensitive and enhanced twofold in HT fibroblasts. In contrast, cellular responses induced by bradykinin, endothelin-1, and angiotensin II (all PTX insensitive) were similar in NT and HT cells. Formation of cAMP induced by stimulation of Gs with isoproterenol was identical in NT and HT cells. Western blot analysis yielded no evidence for an overexpression of G alpha i2, G alpha i3, G beta 2, and G beta 4. Furthermore, sequencing of cDNAs encoding for the ubiquitously expressed PTX-sensitive G protein subunits G alpha i2, G alpha i3, G beta 1, and G beta 2 from NT and HT cell lines yielded no evidence for mutations in these genes. Although the molecular mechanisms remain to be defined, these data support the concept of a selective enhancement of signal transduction via PTX-sensitive G proteins in essential hypertension.
Institut für Pharmakologie, Universitätsklinikum Essen (Germany).
F
F
Pietruck
A
A
Moritz
M
M
Montemurro
A
A
Sell
S
S
Busch
D
D
Rosskopf
S
S
Virchow
H
H
Esche
N
N
Brockmeyer
K H
KH
Jakobs
W
W
Siffert
D001483:Base Sequence; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D003001:Cloning, Molecular; D000242:Cyclic AMP / Q000096:biosynthesis; D004247:DNA / Q000096:biosynthesis; D005347:Fibroblasts / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D006973:Hypertension / Q000503:physiopathology*; D015544:Inositol 1,4,5-Trisphosphate / Q000096:biosynthesis; D007425:Intracellular Membranes / Q000378:metabolism; D009154:Mutation; D009994:Osmolar Concentration; D011956:Receptors, Cell Surface / Q000502:physiology; D012016:Reference Values; D015398:Signal Transduction; D012867:Skin / Q000166:cytology
D011956:Receptors, Cell Surface; D015544:Inositol 1,4,5-Trisphosphate; D004247:DNA; D000242:Cyclic AMP; D019204:GTP-Binding Proteins; D002118:Calcium