Degradation fragments of L1 antigen enhance tyrosine hydroxylase-immunoreactive neurite outgrowth in mesencephalic cell culture.
The L1 antigen has been implicated in adhesion events controlling axonal elongation during formation of major fiber tracts, and promotes neurite outgrowth in culture. It is possible that injury of brain tissue causes neuronal surface molecules such as L1 antigen to be shed, and degradation fragments may therefore be present adjacent to the damage. These L1 fragments might then influence regeneration or injury-induced growth. We have evaluated neurite outgrowth from tyrosine hydroxylase-positive (TH +) E13 mesencephalic neurons grown in vitro on a substrate of mouse L1 antigen and on L1 degradation fragments separated by molecular weight. Mouse myelin-associated glycoprotein (MAG), laminin, poly-D-lysine, and fetal calf serum served as control substrates. L1 antibodies were added to one set of cultures (experimental), and compared to control cultures containing normal rabbit serum. After 3 days in vitro, the cultures were stained using an antibody against TH, and the length of the TH + neurites was measured by computer-assisted image analysis in a double-blind fashion. TH+ neurites were significantly longer when grown on L1 antigen, as well as on L1 degradation fragments, as compared to the control substrates. As compared to control normal rabbit serum, L1 antibodies eliminated the neurite-promoting effect of both the L1 substrate and the L1 degradation products. In addition, L1 substrates promoted clustering of TH + cells and the formation of loose bundles of TH + neurites. It is suggested that the L1 substrates influence TH-immunoreactive neurite outgrowth, at least partially, through indirect effects on glial cells.(ABSTRACT TRUNCATED AT 250 WORDS)
Poltorak
M
M
Preclinical Neuroscience Section, NIMH Neuroscience Center at St. Elizabeths, Washington, DC 20032.
Williams
J R
JR
Freed
W J
WJ
D000818:Animals; D015153:Blotting, Western; D002135:Calcium-Binding Proteins / Q000502:physiology; D015816:Cell Adhesion Molecules, Neuronal / Q000494:pharmacology / Q000502:physiology*; D002478:Cells, Cultured; D004622:Embryo, Mammalian; D007150:Immunohistochemistry; D039841:Leukocyte L1 Antigen Complex; D008636:Mesencephalon / Q000166:cytology / Q000187:drug effects / Q000201:enzymology / Q000502:physiology*; D051379:Mice; D016501:Neurites / Q000187:drug effects / Q000201:enzymology / Q000502:physiology* / Q000648:ultrastructure; D009474:Neurons / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D010446:Peptide Fragments / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D014446:Tyrosine 3-Monooxygenase / Q000032:analysis*
D002135:Calcium-Binding Proteins; D015816:Cell Adhesion Molecules, Neuronal; D039841:Leukocyte L1 Antigen Complex; D010446:Peptide Fragments; D014446:Tyrosine 3-Monooxygenase