Fetal rat septal cells adhere to and extend processes on basement membrane, laminin, and a synthetic peptide from the laminin A chain sequence.
Responses of rat embryonic septal cells to reconstituted basement membrane, laminin, and laminin A chain-derived synthetic peptides were studied in culture. Dissociated fetal E16/17 septal cells were grown for three days on differently coated plastic substrata. Reconstituted basement membrane (Matrigel), laminin, and a 19-amino acid synthetic peptide CSRARKQAASIKVAVSADR-NH2 (PA22-2) from the laminin A chain sequence mediated cell-substratum adhesion and promoted neurite outgrowth. In contrast, cells did not attach to or form processes on uncoated plastic or on plastic substrata coated with synthetic, laminin-derived control peptides. Polyethylenimine (PEI) supported the adhesion and survival of fetal septal cells; however, when laminin was added to the medium during cell plating or 18 hr afterward, a dose-dependent increase was observed in neurite outgrowth of cells attached to this substratum. Cells grown for 6 days on PEI in the presence of laminin showed a determined increase in the number of cholinergic neurons as marked by acetylcholinesterase staining. These data suggest that the subpopulation of cholinergic septal neurons present in the septal cells studied here were also responding to laminin. The results of this in vitro study suggest potential uses for basement membrane, laminin, or synthetic peptides, such as PA22-2, in fetal septal grafts to enhance regeneration in the damaged septo-hippocampal system.
Jucker
M
M
Laboratory of Cellular and Molecular Biology, National Institute on Aging, National Institutes of Health, Baltimore, Maryland 21224.
Kleinman
H K
HK
Ingram
D K
DK
D000110:Acetylcholinesterase / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001485:Basement Membrane / Q000187:drug effects / Q000378:metabolism*; D001921:Brain / Q000166:cytology / Q000378:metabolism*; D002448:Cell Adhesion; D003094:Collagen / Q000494:pharmacology; D004338:Drug Combinations; D004622:Embryo, Mammalian; D005260:Female; D007797:Laminin / Q000494:pharmacology*; D008969:Molecular Sequence Data; D009474:Neurons / Q000187:drug effects / Q000201:enzymology; D010441:Peptide Chain Elongation, Translational / Q000187:drug effects*; D010455:Peptides / Q000494:pharmacology*; D011247:Pregnancy; D011509:Proteoglycans / Q000494:pharmacology; D051381:Rats; D011916:Rats, Inbred F344
D004338:Drug Combinations; D007797:Laminin; D010455:Peptides; D011509:Proteoglycans; C059485:matrigel; C070425:PA22-2; D003094:Collagen; D000110:Acetylcholinesterase