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Effective gene transfer of lacZ and P0 into Schwann cells of P0‐deficient mice
Author(s) -
Guénard Véronique,
Schweitzer Beat,
Flechsig Eckhard,
Hemmi Silvio,
Martini Rudolf,
Suter Ueli,
Schachner Melitta
Publication year - 1999
Publication title -
glia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.954
H-Index - 164
eISSN - 1098-1136
pISSN - 0894-1491
DOI - 10.1002/(sici)1098-1136(19990115)25:2<165::aid-glia7>3.0.co;2-l
Subject(s) - biology , myelin , schwann cell , lac operon , viral vector , sciatic nerve , genetic enhancement , microbiology and biotechnology , gene , gene expression , pathology , anatomy , central nervous system , genetics , neuroscience , medicine , recombinant dna
Mutations in the gene encoding for the myelinating Schwann cell protein P0 have been linked to inherited peripheral neuropathies, including the Charcot‐Marie‐Tooth type 1B disease (CMT1B) and Déjérine‐Sottas syndrome (DSS). Recently generated mice deficient in the P0 gene ( P0 −/− mice) resemble cases of CMT1B and DSS with impaired myelin dosage (Martini et al., 1995a). Potential approaches to treat such diseases include the introduction of the normal gene in the nerves of strongly affected patients. In the present study we used P0 −/− mice to evaluate the efficiency of a replication‐defective, E1‐deleted adenovirus vector carrying the lacZ (Ad‐RSV‐ lacZ ) or P0 (Ad‐RSV‐ P0 ) gene to infect abnormally myelinating Schwann cells. The Ad‐RSV‐ lacZ vector suspension was injected into the left sciatic nerve of P0 −/− mice and the nerves examined for β‐galactosidase activity by X‐gal histochemistry. Contralateral nerves injected with vehicle solution or non‐injected served as controls. β‐galactosidase activity was detected in nerves injected with the Ad‐RSV‐ lacZ vector up to 2 weeks post‐injection. Immunosuppressing the mice with FK506 to decrease the infiltration of activated T‐cells in infected nerves lengthened β‐galactosidase activity to 8 weeks, the longest time point examined. Ultrastructural analysis indicated that X‐gal crystals were present mostly in abnormally myelinating Schwann cells. These findings demonstrate that an adenovirus vector can successfully infect Schwann cells in P0 −/− mice and expression can be maintained for several weeks. The Ad‐RSV‐ P0 suspension was then injected in the sciatic nerve of immunosuppressed P0 −/− mice. Two and four weeks post‐injection both P0 mRNA and protein could be detected by in situ hybridization and Western blotting in some of the nerves. Furthermore, P0 protein expression was observed in myelin‐like structures and onion bulb‐like cells by immunohistochemistry. These results indicate that Schwann cells in P0 −/− mice can be induced to produce P0 protein after gene transfer. Genetic repair of abnormal Schwann cells by using adenovirus vectors might be a possible technique to treat animal models of inherited peripheral neuropathies. GLIA 25:165–178, 1999. © 1999 Wiley‐Liss, Inc.