Abstract
Gene transfer has been used to examine the role of putative neurotransmitters in the nucleus tractus solitarii (NTS). Most such studies used adenovirus vector-mediated gene transfer although adenovirus vector transfects both neuronal and non-neuronal cells. Successful transfection in the NTS has also been reported with lentivirus as the vector. Feline immunodeficiency virus (FIV), a lentivirus, may preferentially transfect neurons and could be a powerful tool to delineate physiological effects produced by altered synthesis of transmitters in neurons. However, it has not been studied in NTS. Therefore, we sought to determine whether FIV transfects rat NTS cells and to define the type of cell transfected. We found that injection of FIV encoding LacZ gene (FIVLacZ) into the NTS led to transfection of numerous NTS cells. Injection of FIVLacZ did not alter immunoreactivity (IR) for neuronal nitric oxide synthase, which we have shown resides in NTS neurons. A majority (91.7 ± 3.9%) of transfected cells contained IR for neuronal nuclear antigen, a neuronal marker; 2.1 ± 3.8% of transfected cells contained IR for glial fibrillary acidic protein, a glial marker. No transfected neurons or fibers were observed in the nodose ganglion, which sends afferents to the NTS. We conclude that FIV almost exclusively transfects neurons in the rat NTS from which it is not retrogradely transported. The cell-type specificity of FIV in the NTS may provide a molecular method to study local physiological functions mediated by potential neurotransmitters in the NTS.
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Abbreviations
- Ad:
-
Adenovirus vector
- AdLacZ:
-
Adenovirus vector with LacZ gene
- βGal:
-
β galactosidase
- Ce:
-
Central subnucleus
- CMV promotor:
-
Cytomegalovirus promotor
- DMV:
-
Dorsal motor nucleus of vagus
- FITC:
-
Fluorescein-5-isothiocyanate
- FIV:
-
Feline immunodeficiency virus vector
- FIVLacZ:
-
FIV vector with LacZ gene
- Gr:
-
Gracilus nucleus
- GFAP:
-
Glial fibrillary acidic protein
- LacZ:
-
Gene encodes β galactosidase
- NeuN:
-
Neuronal nuclear antigen
- NG:
-
Nodose ganglion
- Nnos:
-
Neuronal nitric oxide synthase
- PGP9.5:
-
Protein gene product 9.5
- NTS:
-
Nucleus tractus solitarii
- PBS:
-
Phosphate buffered saline
- RRX:
-
Rhodamine red X
- Tr:
-
Tractus solitarius
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Acknowledgments
This work was funded in part by NIH Grant R01 HL 59593 (to W. T. Talman), NIH Grant R01 HL 088090 (to L. H. Lin and W. T. Talman), and a VA Merit Review Tab 14 (to W. T. Talman).
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Lin, L.H., Langasek, J.E., Talman, L.S. et al. Feline Immunodeficiency Virus as a Gene Transfer Vector in the Rat Nucleus Tractus Solitarii. Cell Mol Neurobiol 30, 339–346 (2010). https://doi.org/10.1007/s10571-009-9456-5
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DOI: https://doi.org/10.1007/s10571-009-9456-5