High transgene expression by lentiviral vectors causes maldevelopment of Purkinje cells in vivo

Cerebellum. 2010 Sep;9(3):291-302. doi: 10.1007/s12311-010-0161-1.


Lentiviral vectors are promising as gene-transfer vehicles for gene therapy targeted to intractable brain diseases. Although lentiviral vectors are thought to exert little toxicity on infected cells, the adverse influence of viral infection on vulnerable developing neurons has not been well studied. Here, we examined whether lentiviral vector infection and subsequent transgene expression affected the morphological and functional maturation of vigorously developing cerebellar Purkinje cells in vivo. Lentiviral vectors expressing GFP under the control of the murine stem cell virus (MSCV) promoter were injected into the cerebellar cortex of neonatal rat pups. Three weeks after treatment, GFP-expressing Purkinje cells were compared with control Purkinje cells from phosphate-buffered saline-injected rats. Analysis of the dendritic tree showed that total dendrite length in GFP-expressing Purkinje cells was almost 80% that in control Purkinje cells. Electrophysiological examination showed that short-term synaptic plasticity at parallel fiber-Purkinje cell synapses and climbing fiber-Purkinje cell synapses was significantly altered in GFP-expressing Purkinje cells. In contrast, maldevelopment of infected Purkinje cells was substantially attenuated when lentiviral vectors with much weaker promoter activity were used. These results suggest that the maldevelopment of Purkinje cells was mainly caused by subsequent expression of a high amount of GFP driven by the strong MSCV promoter. Thus, the use of lentiviral vectors carrying a strong promoter may require particular precautions when applying them to neurological disorders of infants.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Genetic Therapy / methods*
  • Genetic Vectors / adverse effects*
  • Genetic Vectors / genetics
  • Green Fluorescent Proteins / genetics*
  • Immunohistochemistry
  • Lentivirus
  • Lentivirus Infections / genetics
  • Lentivirus Infections / pathology*
  • Microscopy, Confocal
  • Patch-Clamp Techniques
  • Promoter Regions, Genetic
  • Purkinje Cells / pathology*
  • Purkinje Cells / virology*
  • Rats
  • Rats, Wistar
  • Transgenes


  • Green Fluorescent Proteins