Herpes simplex virus (HSV) latency in neurons remains poorly understood, and the heterogeneity of the sensory nervous system complicates mechanistic studies. In this study, we used primary culture of adult trigeminal ganglion (TG) mouse neurons in microfluidic devices, and an model, to examine the subtypes of sensory neurons involved in HSV latency. HSV-infected neurofilament heavy-positive neurons (NefH) were more likely to express Latency-Associated Transcripts (LATs) relative to infected neurofilament heavy-negative neurons (NefH). This differential expression of the LAT promoter correlated with differences in HSV-1 early infection that manifested as differences in the efficiency with which HSV particles reached the cell body following infection at the distal axon. , we further identified a specific subset of NefH neurons which co-expressed Calcitonin Gene Related Peptide α (NefH CGRP) as the sensory neuron subpopulation with the highest LAT promoter activity following HSV-1 infection. Finally, an early-phase reactivation assay showed HSV-1 reactivating in NefH CGRP neurons, although other sensory neuron subpopulations were also involved. Together, these results show that sensory neurons expressing neurofilaments exhibit enhanced LAT promoter activity. We hypothesize that reduced efficiency of HSV-1 invasion at an early phase of infection may promote efficient establishment of latency in NefH neurons due to initiation of the antiviral state preceding arrival of the virus at the neuronal cell body. While the outcome of HSV-1 infection of neurons is determined by a broad variety of factors neuronal subtypes are likely to play differential roles in modulating the establishment of latent infection. Two pivotal properties of HSV-1 make it a successful pathogen. Firstly, it infects neurons which are immune-privileged. Secondly, it establishes latency in these neurons. Together, these properties allow HSV to persist for the lifetime of its host. Neurons are diverse and highly organized cells, with specific anatomical, physiological and molecular characteristics. Previous work has shown that establishment of latency by HSV-1 does not occur equally in all types of neurons. Our results show that the kinetics of HSV infection and levels of latency-related gene expression differ in certain types of neurons. The neuronal subtype infected by HSV is therefore a critical determinant of the outcome of infection and latency.