Herpes simplex (HSV) egress at synapses

单纯疱疹病毒1 （HSV-1）是α疱疹病毒亚家族的一员，是极少数能自然有效地利用哺乳动物神经系统的病毒之一，因此进化出神经元特异性机制，与高度特化的神经细胞生物学相互作用。在典型的病程中，α疱疹病毒感染并在周围神经系统（PNS）中建立潜伏期。在裂解复制过程中，如潜伏期再激活后，子代病毒颗粒扩散回外周组织，引起复发性疱疹性病变，也可扩散到中枢神经系统（CNS）。1型单纯疱疹病毒侵袭中枢神经系统可引起严重和衰弱性疱疹脑炎，或可无症状/亚临床。一旦进入中枢神经系统，单纯疱疹病毒-1可能引发神经炎症，并导致神经退行性疾病的发展。为了更好地了解HSV-1扩散到中枢神经系统的分子和细胞机制，我们提出了一个创新的方法开发/生物工程目标：开发方法直接成像神经元突触或附近的病毒颗粒胞外分泌。这将涉及在有图案的基质上培养初级神经元，以诱导在确定的位置和方向上形成突触。我们将把这种方法与我们已经建立的活细胞病毒胞外作用方法结合起来，以便能够成像和研究突触或突触附近参与病毒胞外作用的细胞生物学因素。阐明HSV-1在神经元中使用的基本细胞生物学过程将增加我们对疱疹病毒在神经系统中传播的方式和原因的理解，导致可药物靶点的鉴定和开发更好的病毒性神经病理学治疗方法，并可能为神经元细胞生物学提供基本见解。

Herpes Simplex Virus 1 (HSV-1), a member of the alpha herpesvirus subfamily, is among the very few viruses that naturally and productively exploit the mammalian nervous system, and, accordingly have evolved neuron- specific mechanisms to interact with highly-specialized neuronal cell biology. During the typical course of disease, alpha herpesviruses infect and establish latency in the peripheral nervous system (PNS). During lytic replication, such as following reactivation from latency, progeny virus particles spread back to peripheral tissues, causing recurrent herpetic lesions, and can also spread into the central nervous system (CNS). HSV-1 invasion of the CNS can cause severe and debilitating herpes encephalitis, or can be asymptomatic/sub- clinical. Once in the CNS, HSV-1 may trigger neuroinflammation and contribute to the development of neurodegenerative disease. To better understand the molecular and cellular mechanisms that underlie HSV-1 spread to the CNS, we propose an innovative method development/bioengineering goal: to develop methods to directly image virus particle exocytosis at or near neuronal synapses. This will involve culturing primary neurons on patterned substrates in order to induce synapse formation at defined locations and orientations. We will combine this method with our established live-cell virus exocytosis method to be able to image and study cell biological factors involved in viral exocytosis at or near synapses. Elucidating the basic cell biological processes that HSV-1 uses in neurons will increase our understanding of how and why herpesviruses spread in the nervous system, lead to the identification of druggable targets and development of better therapies for viral neuropathology, and may provide fundamental insights into the cell biology of neurons.