Adenoviral brain-derived neurotrophic factor induces both neostriatal and olfactory neuronal recruitment from endogenous progenitor cells in the adult forebrain
Adenoviral brain-derived neurotrophic factor induces both neostriatal and olfactory neuronal recruitment from endogenous progenitor cells in the adult forebrain
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DOI：
10.1523/jneurosci.21-17-06718.2001
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发表时间：
2001-09-01
影响因子：
5.3
通讯作者：
Goldman, SA
中科院分区：
文献类型：
--
作者：
Benraiss, A;Chmielnicki, E;Goldman, SA
神经前体细胞存在于成年前脑室管膜下，从中产生的神经元对脑源性神经营养因子(BDNF)产生的反应具有更强的成熟性和存活率。为了从内源性祖细胞诱导神经发生，我们通过转导前脑室管膜结构性表达BDNF，在成年脑室管区过表达BDNF。我们构建了巨细胞病毒(CMV)控制下携带BDNF的双顺反子腺病毒和内部核糖体进入位点(IRES)控制下的人源化绿色荧光蛋白(HGFP)。将AdCMV：BDNF：IRES：hGFP(AdBDNF)注射到成年大鼠的侧脑室，18d后用有丝分裂标志物溴脱氧尿苷(BrdU)处理。注射后3周，注射AdBDNF的动物脑脊液中BDNF平均含量为1µg/g，而对照组脑脊液中检测不到BDNF。原位杂交显示BDNF和GFP mRNA仅在室壁表达。在注射AdBDNF的大鼠中，经共聚焦成像证实，嗅球中BrdU(+)-βIII-微管蛋白(+)神经元的数量比AdNull(AdCMV：hGFP)对照组增加了2.4倍。重要的是，AdBDNF相关的神经元向新纹状体的募集也被注意到，治疗诱导的BrdU(+)-Neun(+)-beta III-tuBulin(+)神经元增加到尾壳核。其中许多细胞还表达谷氨酸脱羧酶、Cabindin-D28和DARPP-32(32 kDa的多巴胺和cAMP调节的磷酸蛋白)，这是新纹状体中等刺神经元的标志。这些新生成的神经元在病毒诱导后至少存活了5-8周。因此，一次注射腺病毒BDNF显著增加了新神经元在成年大鼠脑内神经源性和非神经源性部位的募集。编码神经营养因子的病毒载体的脑室内传递和室管膜感染可能是一种可行的策略，可以诱导成人大脑中的常驻祖细胞分化成神经。
Neural progenitor cells persist throughout the adult forebrain subependyma, and neurons generated from them respond to brain-derived neurotrophic factor (BDNF) with enhanced maturation and survival. To induce neurogenesis from endogenous progenitors, we overexpressed BDNF in the adult ventricular zone by transducing the forebrain ependyma to constitutively express BDNF. We constructed a bicistronic adenovirus bearing BDNF under cytomegalovirus (CMV) control, and humanized green fluorescent protein (hGFP) under internal ribosomal entry site (IRES) control. This AdCMV: BDNF: IRES: hGFP (AdBDNF) was injected into the lateral ventricles of adult rats, who were treated for 18 d thereafter with the mitotic marker bromodeoxyuridine (BrdU). Three weeks after injection, BDNF averaged 1 mug/gm in the CSF of AdBDNF-injected animals but was undetectable in control CSF. In situ hybridization demonstrated BDNF and GFP mRNA expression restricted to the ventricular wall. In AdBDNF-injected rats, the olfactory bulb exhibited a >2.4-fold increase in the number of BrdU(+)-beta III-tubulin(+) neurons, confirmed by confocal imaging, relative to AdNull (AdCMV: hGFP) controls. Importantly, AdBDNF-associated neuronal recruitment to the neostriatum was also noted, with the treatment-induced addition of BrdU(+)-NeuN(+)-beta III-tubulin(+) neurons to the caudate putamen. Many of these cells also expressed glutamic acid decarboxylase, cabindin-D28, and DARPP-32 (dopamine and cAMP-regulated phosphoprotein of 32 kDa), markers of medium spiny neurons of the neostriatum. These newly generated neurons survived at least 5-8 weeks after viral induction. Thus, a single injection of adenoviral BDNF substantially augmented the recruitment of new neurons into both neurogenic and non-neurogenic sites in the adult rat brain. The intraventricular delivery of, and ependymal infection by, viral vectors encoding neurotrophic agents may be a feasible strategy for inducing neurogenesis from resident progenitor cells in the adult brain.