Herpes simplex (HSV) egress at synapses

单纯疱疹病毒1（HSV-1）是α疱疹病毒亚科的成员，是自然地和有效地利用哺乳动物神经系统的极少数病毒之一，并且因此已经进化出神经元特异性机制以与高度特化的神经元细胞生物学相互作用。在典型的疾病过程中，α疱疹病毒感染并在周围神经系统（PNS）中建立潜伏期。在裂解性复制期间，例如从潜伏期重新激活后，子代病毒颗粒扩散回外周组织，引起复发性疱疹病变，并且还可以扩散到中枢神经系统（CNS）中。HSV-1侵入CNS可引起严重和使人衰弱的疱疹性脑炎，或可为无症状/亚临床的。一旦进入中枢神经系统，HSV-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传播到CNS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：开发直接在神经元突触处或附近成像病毒颗粒胞吐的方法。这将涉及在图案化基底上培养初级神经元，以在限定的位置和方向诱导突触形成。我们将结合联合收割机这种方法与我们建立的活细胞病毒胞吐的方法，能够图像和研究细胞生物学因素参与病毒胞吐或附近的突触。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如何以及为什么在神经系统中传播的理解，导致识别可药物靶点和开发更好的病毒神经病理学疗法，并可能提供对神经元细胞生物学的基本见解。

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.