Preferential transfection of adult mouse neural stem cells and their immediate progeny in vivo with polyethylenimine.
The subventricular zone of the adult mammalian brain harbors the neural stem cell population with potential neural regeneration and repair capacity. We describe a nonviral technique to preferentially transfect in vivo the adult neural stem cell population and its immediate progeny based on intraventricular injection of PEI/DNA complexes. The transfected population was identified by cellular and ultra-structural evidence showing their proliferating status and expression of the specific markers GFAP and nestin. Stable activation of the lacZ reporter by cre-recombinase transfection in R26R mice demonstrated survival and migration of stem cell derivatives three months after injection. Apoptosis is thought to be the most common fate of the stem cell progeny. Overexpression of Bcl-X(L) increased number and survival time of transduced progenitors and decreased the frequency of cells immunopositive for activated Caspase-3. This method thus provides selective targeting of the stem cell population and should allow an in-depth understanding of their biology.
Lemkine
Gregory F
GF
Laboratoire de Physiologie Générale et Comparée, Muséum National d'Histoire Naturelle, UMR CNRS 8572, Paris, France.
Mantero
Stefano
S
Migné
Carole
C
Raji
Aicha
A
Goula
Daniel
D
Normandie
Priscilla
P
Levi
Giovanni
G
Demeneix
Barbara A
BA
D000818:Animals; D017209:Apoptosis / Q000235:genetics; D001253:Astrocytes / Q000166:cytology / Q000378:metabolism; D001921:Brain / Q000187:drug effects* / Q000254:growth & development* / Q000648:ultrastructure; D053148:Caspase 3; D020169:Caspases / Q000235:genetics; D002455:Cell Division / Q000235:genetics; D019070:Cell Lineage / Q000187:drug effects* / Q000235:genetics; D002465:Cell Movement / Q000235:genetics; D004247:DNA / Q000494:pharmacology; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects* / Q000502:physiology; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D007150:Immunohistochemistry; D019426:Integrases / Q000235:genetics; D007381:Intermediate Filament Proteins / Q000378:metabolism; D051379:Mice; D008815:Mice, Inbred Strains; D008854:Microscopy, Electron; D009419:Nerve Tissue Proteins; D064231:Nestin; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D010957:Plasmids / Q000494:pharmacology; D011094:Polyethyleneimine / Q000494:pharmacology*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics; D013234:Stem Cells / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D014162:Transfection; D014764:Viral Proteins / Q000235:genetics; D051020:bcl-X Protein; D001616:beta-Galactosidase / Q000235:genetics
C494812:Bcl2l1 protein, mouse; D005904:Glial Fibrillary Acidic Protein; D007381:Intermediate Filament Proteins; D009419:Nerve Tissue Proteins; C577485:Nes protein, mouse; D064231:Nestin; D019253:Proto-Oncogene Proteins c-bcl-2; D014764:Viral Proteins; D051020:bcl-X Protein; D011094:Polyethyleneimine; D004247:DNA; C045073:Cre recombinase; D019426:Integrases; D001616:beta-Galactosidase; C505409:Casp3 protein, mouse; D053148:Caspase 3; D020169:Caspases