Lentiviral transduction of murine oligodendrocytes in vivo.
Lentiviral vectors are used widely to direct efficient gene transfer in vivo. We examined cell-specific expression in adult murine white matter after stereotaxic microinjection of four lentiviral constructs. We synthesized vesicular stomatitis virus glycoprotein (VSV-G) pseudotyped lentiviruses with combinations of two promoters, cytomegalovirus (CMV) or myelin basic protein (MBP), and two reporter sequences, cytosolic enhanced green fluorescent protein (eGFP) or a plasma membrane-targeted eGFP (human lymphocyte-specific protein tyrosine kinase [Lck]-eGFP). For all constructs, intracerebral injections to lateral corpus callosum resulted in widespread GFP expression in forebrain white matter glial cells. Intense cellular GFP fluorescence was observed within 3 days after injection and lasted for at least 28 days. The CMV promoter directed GFP expression in multiple glial cell types, whereas the MBP promoter targeted GFP specifically to oligodendrocytes. Expression of the membrane-targeted Lck-eGFP construct distinctly labeled individual myelinating processes of oligodendrocytes. Lentiviral constructs expressing eGFP or Lck-eGFP under the MBP promoter provide excellent visualization of oligodendrocyte morphology in intact white matter, and may prove valuable for delivering additional genes of interest to oligodendrocytes in vivo.
McIver
Sally R
SR
Hope Center for Neurological Disorders, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Lee
Chul-Sang
CS
Lee
Jin-Moo
JM
Green
Steven H
SH
Sands
Mark S
MS
Snider
B Joy
BJ
Goldberg
Mark P
MP
D000818:Animals; D003587:Cytomegalovirus / Q000235:genetics / Q000378:metabolism; D017930:Genes, Reporter / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics; D016086:Lentivirus / Q000235:genetics*; D051379:Mice; D004676:Myelin Basic Protein / Q000235:genetics; D009413:Nerve Fibers, Myelinated / Q000378:metabolism; D009836:Oligodendroglia / Q000166:cytology* / Q000378:metabolism / Q000821:virology*; D011401:Promoter Regions, Genetic / Q000235:genetics; D016548:Prosencephalon / Q000166:cytology* / Q000378:metabolism / Q000821:virology*; D013194:Staining and Labeling / Q000379:methods; D014161:Transduction, Genetic / Q000379:methods*; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics
D004676:Myelin Basic Protein; D014764:Viral Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins