Chowdhury S I, Coats J, Neis R A, Navarro S M, Paulsen D B, Feng J-M
Department of Pathobiological Sciences, School of Veterinary Medicine, Louisiana State University, Baton Rouge, Louisiana 70803, USA.
J Neurovirol. 2010 Nov;16(6):457-65. doi: 10.1007/BF03210851. Epub 2010 Nov 16.
Bovine herpesvirus type 1 (BHV-1) is an important component of the bovine respiratory disease complex (BRDC) in cattle. Following primary intranasal and ocular infection of cattle, BHV-1 establishes lifelong latent infection in trigeminal ganglia (TG). Upon reactivation from latency, the virus is transported from neuronal cell bodies in the TG to projected nerve endings in nose and cornea of latently infected cattle where the virus shedding occurs. This property of BHV-1 plays a significant role in the pathogenesis of BRDC and maintenance of BHV-1 in the cattle population. Recently, we have reported that a glycoprotein E (gE) cytoplasmic tail-truncated BHV-1 (BHV-1 gEAm453) did not reactivate from latency and was not shed in the nasal and ocular secretions of calves and rabbits. Here we describe the methods to establish rabbit primary dorsal root ganglia (DRG) neuron cultures in a microfluidic chamber system and to characterize in vitro anterograde and retrograde axonal transport properties of BHV-1 gE-deleted and BHV-1 cytoplasmic tail-truncated gEAm453 mutant viruses relative to BHV-1 gEAm453-rescued/wild-type viruses. The results clearly demonstrated that whereas the BHV-1 gE-deleted, BHV-1 gEAm453, and BHV-1 gEAm453-rescued/wild-type viruses were transported equally efficiently in the retrograde direction, only the BHV-1 gEAm453-rescued/wild-type virus was transported anterogradely. Therefore, we have concluded that sequences within the BHV-1 gE cytoplasmic tail are essential for anterograde axonal transport and that primary rabbit DRG neuronal cultures in the microfluidic chambers are suitable for BHV-1 neuronal transport studies.
牛疱疹病毒1型（BHV-1）是牛呼吸道疾病综合征（BRDC）的重要组成部分。牛经鼻内和眼部初次感染BHV-1后，该病毒会在三叉神经节（TG）中建立终身潜伏感染。从潜伏状态重新激活后，病毒从TG中的神经元细胞体运输到潜伏感染牛的鼻子和角膜中的投射神经末梢，在此处发生病毒脱落。BHV-1的这一特性在BRDC的发病机制以及牛群中BHV-1的维持方面发挥着重要作用。最近，我们报道了一种糖蛋白E（gE）胞质尾截短的BHV-1（BHV-1 gEAm453）不会从潜伏状态重新激活，也不会在犊牛和兔子的鼻分泌物和眼分泌物中排出。在此，我们描述了在微流控腔室系统中建立兔初级背根神经节（DRG）神经元培养物的方法，以及相对于BHV-1 gEAm453拯救型/野生型病毒，对缺失BHV-1 gE和截短BHV-1胞质尾的gEAm453突变病毒的体外顺行和逆行轴突运输特性进行表征的方法。结果清楚地表明，虽然缺失BHV-1 gE的病毒、BHV-1 gEAm453以及BHV-1 gEAm453拯救型/野生型病毒在逆行方向上的运输效率相同，但只有BHV-1 gEAm453拯救型/野生型病毒能够顺行运输。因此，我们得出结论，BHV-1 gE胞质尾内的序列对于顺行轴突运输至关重要，并且微流控腔室中的兔初级DRG神经元培养物适用于BHV-1神经元运输研究。
