Dibutyryl cyclic AMP stimulates expression of ependymin mRNA and the synthesis and release of the protein into the culture medium by neuroblastoma cells (NB2a/d1).
Northern blot, immunoprecipitation, and gel electrophoretic data demonstrate that the mouse neuroblastoma NB2a/d1 cells express ependymin mRNA and synthesize and release into the culture medium a protein with immunoreactivity and electrophoretic mobility properties identical to ependymin. This is a brain extracellular glycoprotein that has been implicated in the consolidation process of memory formation and neuronal regeneration. In labeling experiments with 35S-methionine, dibutyrylcyclic3',5'-adenosine-monophosphate (dbcAMP) was found to stimulate the expression of ependymin mRNA and the enhanced synthesis and release of ependymin into the culture medium at the same time that dbcAMP stimulation of neurite outgrowth takes place. These results are consistent with the proposed role of the protein in the mechanism of neuronal regeneration and synaptogenesis. The data indicate that the NB2a/d1 cell line is a good model system for studies of the functional properties of ependymin.
Shashoua
V E
VE
Ralph Lowell Laboratories, McLean Hospital, Harvard Medical School, Belmont, Massachusetts 02178.
Nolan
P M
PM
Shea
T B
TB
Milinazzo
B
B
D000595:Amino Acid Sequence; D000818:Animals; D001345:Autoradiography; D015152:Blotting, Northern; D003994:Bucladesine / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel; D051379:Mice; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D009447:Neuroblastoma / Q000378:metabolism*; D011233:Precipitin Tests; D012333:RNA, Messenger / Q000096:biosynthesis*; D013268:Stimulation, Chemical; D014407:Tumor Cells, Cultured
D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; C031632:ependymins; D003994:Bucladesine