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Identifying HSV infected neurons after ocular inoculation
Abstract:
ICR mice were inoculated intracamerally with McKrae strain herpes simplex virus (HSV) followed by intraperitoneal injection with 3H-thymidine. Infected mice were sacrificed after 3 or 4 days and the eyes, trigeminal ganglia (TG) and superior cervical ganglia (SCG) were embedded in glycol methacrylate, sectioned, and dipped for autoradiography. Light microscopy revealed silver grain labeling over neurons in the ipsilateral retina, TG and SCG of infected animals. No labeling of neurons was noted in the contralateral TG or SCG. Since the DNA of mature neurons does not replicate, we interpret these labeled neurons to represent cells with active replication of HSV. This technique allows the study of HSV infection of the nervous system with excellent tissue preservation. Furthermore, it may be used to distinguish those neurons with intrinsic viral synthesis from those harboring virus synthesized at a distant site with subsequent intracellular spread.
Insights
This study shows that herpes simplex virus (HSV) actively replicates within neurons in the nervous system. Autoradiography reveals labeled neurons, indicating viral DNA synthesis in infected mice.
Area of Science:
- Neurovirology
- Infectious Diseases
- Microscopy Techniques
Background:
- Herpes simplex virus (HSV) is a significant human pathogen.
- Understanding HSV neurotropism and replication is crucial for developing effective treatments.
- Current methods for studying viral replication in neural tissues have limitations.
Purpose of the Study:
- To develop and validate a novel autoradiographic technique for studying HSV replication in neural tissues.
- To investigate the sites of HSV replication within the nervous system of infected mice.
- To differentiate between neurons actively synthesizing HSV and those harboring virus from elsewhere.
Main Methods:
- Intracamercal inoculation of ICR mice with McKrae strain HSV.
- Intraperitoneal injection of 3H-thymidine to label replicating DNA.
- Autoradiography of embedded, sectioned eyes, trigeminal ganglia (TG), and superior cervical ganglia (SCG) after 3-4 days.
- Light microscopy analysis of labeled neurons.
Main Results:
- Silver grain labeling was observed over neurons in the ipsilateral retina, TG, and SCG of HSV-infected mice.
- No neuronal labeling was detected in the contralateral TG or SCG.
- Labeled neurons are interpreted as cells actively replicating HSV, as mature neurons do not replicate their own DNA.
Conclusions:
- The developed autoradiographic technique provides excellent tissue preservation for studying HSV infection in the nervous system.
- This method successfully identifies neurons with active HSV replication.
- The technique can distinguish between intrinsic viral synthesis within neurons and virus spread from distant sites.