Oh Y J, Moffat M, Wong S, Ullrey D, Geller A I, O'Malley K L
Department of Anatomy and Neurobiology, Washington University School of Medicine, St. Louis, MO 63110, USA.
Brain Res Mol Brain Res. 1996 Jan;35(1-2):227-36. doi: 10.1016/0169-328x(95)00221-d.
A defective herpes simplex virus-1 (HSV-1) vector system was used to study cell type-specific expression of the tyrosine hydroxylase (TH) gene. HSV-1 particles containing 663 bp (pTHlac 663), 278 bp (pTHlac 278), or 181 bp (pTHlac 181) of the rat TH promoter driving E. coli LacZ were used to infect superior cervical ganglia (SCG: TH-expressing tissue) and dorsal root ganglia (DRG:non-TH-expressing tissue) cultures. One day after infection, expression of beta-galactosidase was visualized by X-gal cytochemistry. Following viral transduction with pTHlac 663 at a multiplicity of infection of 0.2, 14.4% of the SCG neurons were X-gal positive whereas only about 0.9% of DRG neurons were X-gal positive. Infection with either pTHlac278 or 181 resulted in 3-fold more X-gal-positive DRG neurons. These results suggest that (i) the defective HSV-1 vector system may be useful in defining regulatory promoter motifs; (ii) 663 bp of the rat TH promoter contains sufficient information for cell type-specific expression in peripheral nervous system neurons; and (iii) sequences between -278 and -663 contain an element(s) that represses gene expression in non-catecholamingeric neurons.
一种缺陷型单纯疱疹病毒1型(HSV-1)载体系统被用于研究酪氨酸羟化酶(TH)基因的细胞类型特异性表达。含有由大鼠TH启动子驱动大肠杆菌LacZ的663 bp(pTHlac 663)、278 bp(pTHlac 278)或181 bp(pTHlac 181)的HSV-1颗粒被用于感染颈上神经节(SCG:表达TH的组织)和背根神经节(DRG:不表达TH的组织)培养物。感染后一天,通过X-半乳糖苷酶细胞化学法观察β-半乳糖苷酶的表达。以0.2的感染复数用pTHlac 663进行病毒转导后,14.4%的SCG神经元呈X-半乳糖苷酶阳性,而只有约0.9%的DRG神经元呈X-半乳糖苷酶阳性。用pTHlac278或181感染导致DRG神经元中X-半乳糖苷酶阳性的数量增加了3倍。这些结果表明:(i)缺陷型HSV-1载体系统可能有助于确定调控启动子基序;(ii)大鼠TH启动子的663 bp包含在外周神经系统神经元中进行细胞类型特异性表达的足够信息;(iii)-278至-663之间的序列包含一个在非儿茶酚胺能神经元中抑制基因表达的元件。