Feline immunodeficiency virus as a gene transfer vector in the rat nucleus tractus solitarii.
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.
Lin
L H
LH
Department of Neurology, 1191 ML, University of Iowa, Iowa City, IA 52242, USA. li-hsien-lin@uiowa.edu
Langasek
J E
JE
Talman
L S
LS
Taktakishvili
O M
OM
Talman
W T
WT
D000344:Afferent Pathways / Q000166:cytology / Q000201:enzymology; D000818:Animals; D034961:Antigens, Nuclear / Q000235:genetics; D001370:Axonal Transport / Q000502:physiology; D001931:Brain Mapping / Q000379:methods; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics*; D005904:Glial Fibrillary Acidic Protein / Q000235:genetics; D016089:Immunodeficiency Virus, Feline / Q000235:genetics*; D008297:Male; D009419:Nerve Tissue Proteins / Q000235:genetics; D056347:Neuroanatomical Tract-Tracing Techniques / Q000379:methods; D009474:Neurons / Q000166:cytology / Q000201:enzymology / Q000821:virology; D018377:Neurotransmitter Agents / Q000096:biosynthesis*; D009620:Nodose Ganglion / Q000166:cytology / Q000201:enzymology; D051381:Rats; D017207:Rats, Sprague-Dawley; D017552:Solitary Nucleus / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*; D013194:Staining and Labeling / Q000379:methods; D014162:Transfection / Q000379:methods*; D001616:beta-Galactosidase / Q000235:genetics
D034961:Antigens, Nuclear; D005904:Glial Fibrillary Acidic Protein; D009419:Nerve Tissue Proteins; D018377:Neurotransmitter Agents; C546339:Rbfox3 protein, rat; D001616:beta-Galactosidase