Use of genetically modified glial cells overexpressing laminin alpha1-chain peptides in neurite outgrowth studies.
1. C6 glioma cells were transfected with two constructs carrying C-terminal laminin alpha1-chain sequences of 117 and 114 bp length, respectively. These sequences are specifically known to code for peptides which have neurite-promoting activity. 2. The stable expression and secretion of the two peptides was detected by Northern and Western blot analysis. 3. Primary neuronal cultures derived from embryonic mouse forebrain were cocultured with these transfected cells and exhibited a substantial increase in neurite outgrowth and in survival time. Conditioned media from the transfected cells generated similar effects. 4. Organotypic cultures from embryonic mouse brain were used as a second system as being closer to the in vivo situation. Again, coculture of brain slices with transfected cells or treatment with laminin peptide-containing media increased neuronal outgrowth.
Webersinke
G
G
Institute for Molecular Biology, Austrian Academy of Science, Salzburg.
Bauer
H C
HC
Danninger
C
C
Krizbai
I A
IA
Schittny
J C
JC
Thalhamer
J
J
Bauer
H
H
D000818:Animals; D002478:Cells, Cultured; D003001:Cloning, Molecular; D018920:Coculture Techniques; D004622:Embryo, Mammalian; D005910:Glioma; D007797:Laminin / Q000032:analysis / Q000235:genetics* / Q000502:physiology; D051379:Mice; D008815:Mice, Inbred Strains; D016501:Neurites / Q000502:physiology*; D009457:Neuroglia / Q000166:cytology / Q000502:physiology*; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D009924:Organ Culture Techniques; D016133:Polymerase Chain Reaction; D016548:Prosencephalon / Q000166:cytology / Q000196:embryology; D011994:Recombinant Proteins / Q000032:analysis; D015183:Restriction Mapping; D014162:Transfection; D014407:Tumor Cells, Cultured
D007797:Laminin; D011994:Recombinant Proteins; C084098:laminin A