Transduction of protein into rat brainstem using the Hemagglutinating virus of Japan-envelope vector in vivo

Abstract

Viral vectors have been mainly used for the various studies of gene and cell therapies because of their inherent property of high transfection. However, the clinical applications of viral vectors are limited by their cytotoxicity and immunogenicity. Hemagglutinating virus of Japan envelope (HVJ-E) vector we used in this study is a novel non-viral vector which is inactivated and lost the ability to replicate.
We demonstrated the transduction of β-galactosidase (β-gal) as a delivery protein incorporated with HVJ-E into the rat brainstem in vivo. We microinjected HVJ-E/β-gal (n=16)(100nl/site) and control HVJ-E vector (n=13) into the nucleus tractus solitarius of anesthertized rats. The rats were fixed by intracardic perfusion with 2% paraformaldehyde-0.2% glutaraldehyde in phosphate-buffered saline (pH7.4) at 3, 6, 12, 24 hours after microinjection. Brainstems were removed and cryostat sections(40μm) were obtained from the region of the injection and assayed by X-gal staining. Sections from HVJ-E/β-gal-injected rats showed β-gal activities in all cases. Sections from HVJ-E-injected rats showed no staining with X-gal. Histological stainings were assessed by measuring the colored strength in a defined area using the Mac Scope software. It is suggested that the delivery of proteins using HVJ-E vector may introduce a new system of transduction to study a therapeutic neurobiology. [Jpn J Physiol 54 Suppl:S79 (2004)]