Christenson S D, Lake K D, Ooboshi H, Faraci F M, Davidson B L, Heistad D D
Department of Internal Medicine, Cardiovascular Center and Center on Aging, University of Iowa College of Medicine, and Veterans Administration Medical Center, Iowa City 52242, USA.
Stroke. 1998 Jul;29(7):1411-5; discussion 1416. doi: 10.1161/01.str.29.7.1411.
Gene transfer to cerebral blood vessels has been accomplished in rats and dogs by injection of replication-deficient adenovirus into cerebrospinal fluid. In this study we examined transgene expression after injection of adenovirus into the cerebrospinal fluid of mice. Responses were observed in ICR mice and C57BL/6 mice, which are outbred and inbred strains, respectively.
We injected replication-deficient recombinant adenovirus expressing nuclear targeted beta-galactosidase, driven by either the Rous sarcoma virus promoter (AdRSV-betaGal) or the cytomegalovirus promoter (AdCMV-betaGal), into the cisterna magna of anesthetized ICR and C57BL/6 strains of mice. The brains were examined from 1 to 21 days after injection by chemiluminescent enzyme activity assay or histochemical staining.
After injection of AdRSV-betaGal, expression of beta-galactosidase in ICR mice peaked on day 7 and returned to basal by day 14. Expression of beta-galactosidase in C57BL/6 mice was maximal on days 7 to 14 and was minimal by day 21 after injection of AdRSV-betaGal. After injection of AdCMV-betaGal in C57BL/6 mice, peak expression of transgene occurred on day 1 and was greatly diminished by day 3. Transgene expression was observed primarily on the ventral surface of the brain, with preferential expression in leptomeninges and adventitia along the major cerebral arteries of that region.
Injection of recombinant adenovirus in the cisterna magna resulted in transgene expression in leptomeninges and perivascular tissue of cerebral blood vessels in two strains of mice. The CMV promoter elicited rapid but short-lived expression of the transgene, while the RSV promoter elicited slower, more sustained transgene expression. Expression of AdRSV transgene was prolonged in C57BL/6 mice compared with ICR mice. This approach for gene transfer may be useful to study cerebral vascular biology in genetically altered strains of mice.
通过向脑脊液中注射复制缺陷型腺病毒，已在大鼠和犬类中实现了向脑血管的基因转移。在本研究中，我们检测了将腺病毒注射到小鼠脑脊液后转基因的表达情况。分别在远交系ICR小鼠和近交系C57BL/6小鼠中观察了反应。
我们将由劳斯肉瘤病毒启动子（AdRSV-βGal）或巨细胞病毒启动子（AdCMV-βGal）驱动的表达核靶向β-半乳糖苷酶的复制缺陷型重组腺病毒注射到麻醉后的ICR和C57BL/6品系小鼠的小脑延髓池。在注射后1至21天，通过化学发光酶活性测定或组织化学染色检查大脑。
注射AdRSV-βGal后，ICR小鼠中β-半乳糖苷酶的表达在第7天达到峰值，并在第14天恢复到基础水平。注射AdRSV-βGal后，C57BL/6小鼠中β-半乳糖苷酶的表达在第7至14天最高，在第21天最低。在C57BL/6小鼠中注射AdCMV-βGal后，转基因的峰值表达出现在第1天，并在第3天大大降低。转基因表达主要在脑腹侧表面观察到，在该区域主要脑动脉周围的软脑膜和外膜中有优先表达。
在小脑延髓池注射重组腺病毒导致转基因在两种品系小鼠的软脑膜和脑血管周围组织中表达。CMV启动子引发转基因的快速但短暂的表达，而RSV启动子引发较慢、更持久的转基因表达。与ICR小鼠相比，AdRSV转基因在C57BL/6小鼠中的表达延长。这种基因转移方法可能有助于在基因改造的小鼠品系中研究脑血管生物学。
