Receptor-mediated modulation of recombinant neuronal class E calcium channels.

The modulation of a cloned neuronal calcium channel was studied in a human embryonic kidney cell line (HEK293). The HEK293 cells were stably transfected with the alpha1Ed cDNA, containing the pore forming subunit of a neuronal class E calcium channel. Inward currents of 25 +/- 1.9 pA/pF (n = 79) were measured with the cloned alpha1Ed-subunit. The application of the peptide hormone somatostatin, carbachol, ATP or adenosine reduced the amplitude of Ca2+ and Ba2+ inward currents and exhibited a slowing of inactivation. This inhibitory effect by somatostatin was significantly impaired after pre-incubating the transfected cell line with pertussis toxin (PTX). Internal perfusion of the cells with the G-protein-inactivating agent GDP-beta-S or with the permanently activating agent GTP-gamma-S also attenuated the somatostatin effect. The inhibition indicates that modulation of the alpha1Ed-mediated Ca2+ current involves pertussis toxin-sensitive G-proteins. The block of Ca2+ and Ba2+ inward currents by somatostatin is also found in cells expressing a truncated alpha1Ed-subunit which lacks a 129-bp fragment in the C-terminus. This fragment corresponds to the major structural difference between two native human alpha1E splice variants. As somatostatin inhibits inward currents through both, the cloned alpha1Ed- and the truncated alpha1Ed-DEL-subunit, the hormone-mediated modulation is independent from the presence of the 129-bp insertion in the C-terminus.