Adenovirus-mediated G alpha(q)-protein antisense transfer in neurons replicates G alpha(q) gene knockout strategies

Antisense approaches are increasingly used to dissect signaling pathways linking cell surface receptors to intracellular effectors. Here we used a recombinant adenovirus to deliver G-protein alpha(q) antisense into rat superior cervical ganglion (SCG) neurons and neuronal cell lines to dissect Galpha(q)-mediated signaling pathways in these cells. This approach was compared with other Galpha(q) gene knockdown strategies, namely, antisense plasmid and knockout mice. Infection with adenovirus expressing Galpha(q) antisense (Galpha(q)AS AdV) selectively decreased immunoreactivity for the Galpha(q), protein. Expression of other Galpha protein subunits, such as Galpha(oA/B) was unaltered. Consistent with this, modulation of Ca2+ currents by the Galpha(q)-coupled M-1 muscarinic receptor was unchanged. In agreement neurons infected with Galpha(q)AS AdV whereas modulation via the Galpha(oA)-coupled M, muscarinic receptor was Unchanged. In agreement. activation of phospholipase C and consequent mobilization of intracellular Ca2- by UTP receptors k as lost in NG108-15 cells infected with Galpha(q)AS AdV but not in cells infected with the control GFP-expressing, adenovirus. Results obt dned with this recombinant AdV strategy quahtatively and quantitatively replicated results obtained usim, SCG neurons microinjected with Galpha(q) antisense plasmids or SCG neurons from Galpha(q) knockout mice. This combined antisense/recombinant adenoviral approach can therefore be useful for dissecting signal transduction mechanisms in SCG and other neurons. (C) 2002 Elsevier Science Ltd, All rights reserved.