The development of viral vector for in vivo gene-delivery and its applications

体内基因传递病毒载体的研制及其应用

• 批准号: 05558094
• 负责人: SENBA Emiko
• 金额: $ 11.39万
• 依托单位: Wakayama Medical College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-05558094/
• 关键词: pseudorabies virus; transneuronal tracer; gene-delivery; masseter; motor trigeminal nucleus; mesencephalic trigeminal nucleus; retrograde transport; pseudorabies virus; トランスフェクション; 神経細胞; トレーサー; 後根神経節; ラット; Pseudorabies virus; Transneuronal t

Fusion gene of non-essential glycoprotein gG (gX) and lacZ was constructed and pseudorabies virus (PrV) mutants were produced. Characteristics and usefulness of this mutant virus as a transneuronal tracer was investigated by injecting it into masseter or stomach of SD rats. Injection of PrV solution with titer of 1.0*10^7p.f.u./ml was most suitable for the tracing study. Injection of higher titer of viral solution killed the animals within 2 days, indicating the retained virulence of this mutant virus. Although viral infection occurred in motor trigeminal n.neurous in all cases of masseter injection, labeled neurous in the mesencephalic trigeminal n.were very few. Second-or third-order neurons were identified in the locus coeruleus (LC), raphe magnus (RM), medullary reticular formation (IRt), Edinger-Westphal nucleus (EW), tuberal magnocellular n.(TM), lateral hypothalamic n.(LH) and paraventricular hypothalamic n.(PVH). No anterograde tracing was detected in the CNS.Injection of this virus into stomach labeled neurons in the dorsal motor nucleus of N.X (DMN), n.tractus solitarius, n. ambiguus, A5 and PVH.These findings indicate that this virus preferentially infecct somatic and visceral motoneurons and is retrogradely tranported in second-order or third-order neurons in the CNS.

构建了伪狂犬病病毒（PrV）非必需糖蛋白gG（gX）与lacZ的融合基因，并获得了伪狂犬病病毒（PrV）突变株。通过将其注射到SD大鼠的咬肌或胃中，研究了该突变病毒作为跨神经元示踪剂的特性和有效性。注射滴度为1.0* 10^7p.f.u./ ml最适合示踪研究。注射更高滴度的病毒溶液在2天内杀死动物，表明该突变病毒保留了毒力。咬肌注射的三叉神经运动神经均发生病毒感染，但标记神经在三叉神经中脑神经的病例很少。在蓝斑（LC）、中缝大核（RM）、延髓网状结构（IRt）、Edinger-Westphal核（EW）、结节大细胞核（EW）、延髓网状结构（IRt）等核团内均发现二级或三级神经元。(TM)、下丘脑外侧核。(LH)和室旁下丘脑神经（PVH）。将该病毒注入胃内，可标记孤束核、孤束核、孤束核和孤束核的运动背核（DMN）神经元。这些结果表明，该病毒优先感染躯体和内脏运动神经元，并在中枢神经系统的二级或三级神经元中逆行转运。

项目成果

Miguel-Hidalgo,J.J.et al.: "Projections of tachykinin-and glutaminase containing rat retinal ganglion cells" Brain Research Bulletin. 35. 73-84 (1994)

Miguel-Hidalgo, J.J. 等人：“含有大鼠视网膜神经节细胞的速激肽和谷氨酰胺酶的投影”大脑研究通报。

Nemoto K.et al.: "Mutation of low affinity nerve growth factor receptor gene is associated with the hypertensive phenotype in SHR." Neurosci.Lett.210. 69-72 (1996)

Nemoto K.等人：“低亲和力神经生长因子受体基因的突变与 SHR 的高血压表型相关。”

Noguchi,K.et al.: "Substance P induced by peripheral nerve injury in primary afferent neurons and its effect on dorsal column nucleus neurons." Journal of Neuroscience. 15. 7633-7643 (1995)

Noguchi, K. 等人：“初级传入神经元周围神经损伤诱导的 P 物质及其对背柱核神经元的影响。”

Noguchi, K., De Leon, M., Nahin, R.L., Senba, E.and Ruda, M.A.: "Quantification of axotomy-induced alteration of neuropeptide mRNAs in dorsal root ganglion neurons with special reference to neuropeptide Y mRNA and the effects of neonatal capsaicin treatme

Noguchi, K.、De Leon, M.、Nahin, R.L.、Senba, E. 和 Ruda, M.A.：“对背根神经节神经元中轴切术诱导的神经肽 mRNA 改变进行定量，特别是神经肽 Y mRNA 以及

Tokunaga,A et al.: "Effects of local anesthesia on formalin‐induced Fos expression in the rat dorsal horn." Neuroreport. 6. 2301-2304 (1995)

Tokunaga, A 等人：“局部麻醉对大鼠背角福尔马林诱导的 Fos 表达的影响。Neuroreport。”6. 2301-2304 (1995)