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Expression of heterologous proteins in cultured rat hippocampal neurons using the semliki forest virus vector
Author(s) -
Olkkonen V. M.,
Liljeström P.,
Garoff H.,
Simons K.,
Dotti C. G.
Publication year - 1993
Publication title -
journal of neuroscience research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.72
H-Index - 160
eISSN - 1097-4547
pISSN - 0360-4012
DOI - 10.1002/jnr.490350412
Subject(s) - semliki forest virus , golgi apparatus , biology , microbiology and biotechnology , heterologous , viral vector , hippocampal formation , immunofluorescence , synaptic vesicle , vesicle , virology , endoplasmic reticulum , recombinant dna , biochemistry , immunology , neuroscience , antibody , membrane , rna , gene
The Semliki Forest virus expression vector (Liljeström and Garoff: Bio/Technology 9:1356‐1361, 1991) was tested in cultured rat hippocampal neurons using two Madin‐Darby canine kidney (MDCK) cell membrane‐associated proteins as reporters: rab8, a small GTP ase involved in post‐Golgi vesicle transport, and VIP21, an integral membrane protein of caveolae, trans‐Golgi network, and post‐Golgi vesicles. Expression of the c‐ myc epitope‐tagged proteins was visualized by immunofluorescence microscopy. The proteins were first detected in neurons after 3–4 hr infection by the recombinant viruses. The infection efficiency on neurons was high: after 6 hr infection at a multiplicity of one, 50–60% of the cells expressed the reporter proteins. The neurons tolerated the in fection well up to 8 hr. Their polarized organization was not disturbed, as judged from morphology and from distribution of the dendritic MAP2 and axonal synaptophysin marker proteins. The Semliki Forest virus vector thus seems suitable for short‐term expression of proteins in cultured neurons. © 1993 Wiley‐Liss, Inc.