Alpha Herpesvirus Egress and Spread from Neurons Uses Constitutive Secretory Mechanisms Independent of Neuronal Firing Activity

Anthony E. Ambrosini; Nikhil Deshmukh; Michael J. Berry; Lynn W. Enquist; Ian B. Hogue

225 To determine whether optogentically-controlled neuronal activity affects spread of 226 PRV, we prepared tri-chambers with SCG neurons in the left soma chamber, and 227 transduced with an AAV vector to express ChR2-mCherry. We screened neurons 228 by fluorescence microscopy to ensure ChR2-mCherry expression in >80% of 229 cells. At 4 weeks in vitro, we seeded PK15 recipient cells in the right neurite 230 compartment, infected with PRV Becker, and pulsed with blue light beginning at 231 1 hpi ( Fig 4E) . In the experimental condition, where neurons were transduced 232 with ChR2 and pulsed with blue light, neurite compartment titers were slightly 233 less than controls at 17 and 20 hpi, but not significantly different by 24 hpi 234 (p>0.05) (Fig 4F) .