Alpha herpesvirus exocytosis from neuron cell bodies uses constitutive secretory mechanisms, and egress and spread from axons is independent of neuronal firing activity
Fig 7
Neuronal activity does not correlate with PRV spread from neurons: optogenetics approach.
(A-B) SCG neurons were seeded onto a multi-electrode array, transduced with an AAV vector expressing ChR2-mCherry. (A) Scale bar is 500μm. (B) Scale bar is 100μm. (C) Extracellular electrical recordings of SCG neurons exhibiting spontaneous spiking activity (no stimulus), or evoked activity synchronized to 12 Hz blue light pulses. (D) Superimposed spikes (n = 100) demonstrating increases in spike peak-to-peak amplitude with optogenetic stimulation. (E) Schematic of modified Campenot tri-chamber. SCG neurons were seeded in the left soma compartment and extended axons under the chamber walls to the right neurite compartment. A detector cell monolayer (PK15) was then added to the neurite compartment. SCGs were transduced with a ChR2 AAV and exposed to pulsed blue light, as indicated. PRV infection was initiated in the soma compartment at a multiplicity of infection of 10 or 100, as indicated. (F) Virus titer measured in neurite chambers by serial dilution plaque assay. No significant difference (p>0.05) at 24 hpi.