Pseudorabies virus expressing enhanced green fluorescent protein: A tool for in vitro electrophysiological analysis of transsynaptically labeled neurons in identified central nervous system circuits
| العنوان: | Pseudorabies virus expressing enhanced green fluorescent protein: A tool for in vitro electrophysiological analysis of transsynaptically labeled neurons in identified central nervous system circuits |
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| المؤلفون: | Bruce W. Banfield, Bret N. Smith, Christine L. Wilcox, Gary E. Pickard, Lynn W. Enquist, F. Edward Dudek, Cynthia A. Smeraski |
| المصدر: | Proceedings of the National Academy of Sciences. 97:9264-9269 |
| بيانات النشر: | Proceedings of the National Academy of Sciences, 2000. |
| سنة النشر: | 2000 |
| مصطلحات موضوعية: | Central Nervous System, Male, Retinal Ganglion Cells, Green Fluorescent Proteins, Central nervous system, Pseudorabies, Synaptic Transmission, Retinal ganglion, Green fluorescent protein, chemistry.chemical_compound, Cricetinae, medicine, Animals, Humans, Multidisciplinary, Mesocricetus, biology, Suprachiasmatic nucleus, Retinal, Biological Sciences, biology.organism_classification, Herpesvirus 1, Suid, Virology, Cell biology, Luminescent Proteins, medicine.anatomical_structure, nervous system, Retinal ganglion cell, chemistry, Synapses, Axoplasmic transport, sense organs |
| الوصف: | Physiological properties of central nervous system neurons infected with a pseudorabies virus were examined in vitro by using whole-cell patch-clamp techniques. A strain of pseudorabies virus (PRV 152) isogenic with the Bartha strain of PRV was constructed to express an enhanced green fluorescent protein (EGFP) from the human cytomegalovirus immediate early promoter. Unilateral PRV 152 injections into the vitreous body of the hamster eye transsynaptically infected a restricted set of retinorecipient neurons including neurons in the hypothalamic suprachiasmatic nucleus (SCN) and the intergeniculate leaflet (IGL) of the thalamus. Retinorecipient SCN neurons were identified in tissue slices prepared for in vitro electrophysiological analysis by their expression of EGFP. At longer postinjection times, retinal ganglion cells in the contralateral eye also expressed EGFP, becoming infected after transsynaptic uptake and retrograde transport from infected retinorecipient neurons. Retinal ganglion cells that expressed EGFP were easily identified in retinal whole mounts viewed under epifluorescence. Whole-cell patch-clamp recordings revealed that the physiological properties of PRV 152-infected SCN neurons were within the range of properties observed in noninfected SCN neurons. Physiological properties of retinal ganglion cells also appeared normal. The results suggest that PRV 152 is a powerful tool for the transsynaptic labeling of neurons in defined central nervous system circuits that allows neurons to be identified in vitro by their expression of EGFP, analyzed electrophysiologically, and described in morphological detail. |
| تدمد: | 1091-6490 0027-8424 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::12cd65831d65edde544753ef90559d63 https://doi.org/10.1073/pnas.97.16.9264 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....12cd65831d65edde544753ef90559d63 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 10916490 00278424 |
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