Glucagon increases intracellular free calcium in a distal tubular cell line

Glucagon (Glu) influences renal tubular function and growth, although the signal transduction of Glu in the kidney still remains obscure. Rabbit cortical tubules were transformed by the pSV-neo3 gene to make a homogeneous cell colony, which responded to vasopressin but not to parathyroid hormone. The [Ca2+]i of the cells at the 9-10th passages was measured by the fluorescence indicator, fura-2. The [Ca2+]i was increased by Glu (10(-8) M) or bradykinin (10(-8) M), between which heterologous desensitization was observed. The Glu range of 10(-14) to 10(-6) M) significantly increased [Ca2+]i, while cAMP was not produced at any dose of Glu. Since the ranges of doses were from physiological to pharmacological, two concentrations of 10(-13) and 10(-8) M were employed to investigate the mechanisms. Glu at 10(-13) M led to a sustained rise in [Ca2+]i, which was completely blocked by external EGTA (5 mM, Ca-free solution). Glu at 10(-8) M provided a similar level of peak and sustained rise in [Ca2+]i, the sustained phase of which was blunted in Ca-free solution. Inositol tri/tetra phosphates were significantly increased by 10(-8) M, but not by 10(-13) M Glu. These data suggest that [Ca2+]i elevation is a major component of Glu-induced second messengers in the physiological and pharmacological range of doses of Glu, and that there might be two classes of pathways leading to increase in [Ca2+]i in transformed rabbit cortical collecting tubule cells.