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.

构建了非必需糖蛋白GG（GX）和LACZ的融合基因，并产生了伪酸病毒（PRV）突变体。该突变病毒作为跨神经元示踪剂的特征和有用性通过将其注射到SD大鼠的蜂蜜或胃中来研究。用1.0*10^7p.f.u./ml注入PRV溶液最适合追踪研究。注射较高的病毒溶液的较高滴度在2天内杀死了动物，表明该突变病毒的毒力保留了。尽管在所有咬合注射的情况下，在运动三叉神经上都发生了病毒感染，但在中脑三叉神经中被标记为神经。在基因座（LC），Raphe Magnus（RM），髓质网状形成（IRT），Edinger-Westphal核（EW），结节巨细胞n。（TM）中鉴定出第二阶神经元（LC），Raphe Magnus（RM）（RM）（RM）（IRT）。 （LH）和旁脊髓下丘脑n。（PVH）。在CN中未检测到未检测到该病毒在N.X（DMN）背运动核中的胃中的射击。 Ambiguus，A5和PVH。这些发现表明，该病毒优先地是在CNS中以二阶或三阶神经元的逆转来逆转体运动神经元和内脏运动神经元。

项目成果

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Kashiba, H., Noguchi, K., Ueda, Y.and Senba E.: "Neuropeptide Y and galanin are coexpressed in rat large type A sensory neurons after peripheral transection" Peptides. 15. 411-416

Kashiba, H.、Noguchi, K.、Ueda, Y. 和 Senba E.：“神经肽 Y 和甘丙肽在外周横断后在大鼠大 A 型感觉神经元中共表达”肽。