Can anbody make some comments on the antisense delivery methods to neuron cells? Thanks.

Adenovirus mediated delivery has been successfully used to deliver antisense into rat superior cervical ganglion (SCG) neurons and neuronal cell lines

see
Neuropharmacology
Volume 42, Issue 7, June 2002, Pages 950-957
Adenovirus-mediated G(alpha)q -protein antisense transfer in neurons replicates G(alpha)q gene knockout strategies

Abstract

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 G(alpha)q-mediated signaling pathways in these cells. This approach was compared with other G(alpha)q gene knockdown strategies, namely, antisense plasmid and knockout mice. Infection with adenovirus expressing Ggreek small letter alphaq antisense (G(alpha)qAS AdV) selectively decreased immunoreactivity for the G(alpha)q protein. Expression of other G(alpha) protein subunits, such as G(alpha)oA/B, was unaltered. Consistent with this, modulation of Ca2+ currents by the G(alpha)q-coupled M1 muscarinic receptor was severely impaired in neurons infected with Gg(alpha)qAS AdV whereas modulation via the G(alpha)oA-coupled M4 muscarinic receptor was unchanged. In agreement, activation of phospholipase C and consequent mobilization of intracellular Ca2+ by UTP receptors was lost in NG108-15 cells infected with G(alpha)qAS AdV but not in cells infected with the control GFP-expressing adenovirus. Results obtained with this recombinant AdV strategy qualitatively and quantitatively replicated results obtained using SCG neurons microinjected with G(alpha)q antisense plasmids or SCG neurons from G(alpha)q knockout mice. This combined antisense/recombinant adenoviral approach can therefore be useful for dissecting signal transduction mechanisms in SCG and other neurons.


SEE also
Am J Physiol Heart Circ Physiol 274: H719-H727, 1998;

Attenuation of ANG II actions by adenovirus delivery of AT1 receptor antisense in neurons and SMC

Di Lu, Hong Yang, and Mohan K. Raizada

Department of Physiology, University of Florida, Gainesville, Florida 32610

Both central and peripheral renin-angiotensin systems (RAS) are important in the development and establishment of hypertension. Thus, introducing genes relevant to RAS into neuronal and vascular smooth muscle (VSM) cells, two major targets for angiotensin (ANG) II action, is a prerequisite in considering a gene therapy approach for the control of ANG-dependent hypertension. In this study, we explored the use of adenoviral (Ad) vector to transfer AT1 receptor antisense cDNA (AT1R-AS) into neuronal and VSM cells with the anticipation of attenuation of ANG II-mediated cellular actions. Incubation of neurons and VSM cells with viral particles containing AT1R-AS (Ad-AT1R-AS) resulted in a robust expression of AT1R-AS in a majority (~80%) of the cells. The expression was persistent for at least 28 days and was associated with decreases in the immunoreactive AT1 receptor protein and the maximal binding for AT1 receptor in a time- and dose-dependent manner in both cell types. ANG II stimulation of [3H]thymidine incorporation in VSM cells and norepinephrine transporter gene expression in neuronal cells were attenuated by Ad-AT1R-AS infection. Uninfected cells or cells infected with adenovirus particles containing a mutant AT1 receptor sense cDNA showed no effects on either AT1 receptor or on attenuation of ANG II's cellular affects. These observations show, for the first time, that adenovirus can be used to deliver AT1 receptor mutant sense and antisense cDNAs into two major ANG II target tissues. This consequently influences AT1 receptor-mediated cellular actions of ANG II.