Overexpression of human transferrin in two oligodendroglial cell lines enhances their differentiation.
We have previously demonstrated that the addition of apotransferrin (aTf) to oligodendroglial cell (OLGc) primary cultures accelerates their maturation. Cells treated with aTf developed a multipolar morphology and displayed increased expression of mature OLGc markers. In this work, we studied the effect of Tf overexpression in two OLGc lines, N19 and N20.1. The former cells exhibit characteristics of OLGc precursors (O2A), while N20.1 cells express markers of more mature OLGcs. Using the complete cDNA of the human Tf gene, we obtained clones overexpressing Tf in both cell lines. These clones were evaluated for the expression of OLGc differentiation markers. In agreement with our previous results, we found that in the cells overexpressing Tf, there was an increased O(4), GC, and MBP immunoreactivity. To study the myelinogenic potential of these cells, we co-cultured N19 and N20.1 Tf-transfected cells together with cortical neurons. There was a dramatic increase in the morphological differentiation of the OLGcs accompanied by enhanced GC and MBP expression. The OLGcs appeared to establish contact with neurites and extend their processes along them. Only two MBP isoforms were detected in Tf-overexpressing clones, while all the isoforms were present in the co-cultures, suggesting that there was a modulation of MBP expression by neurons. Concomitantly, we found an increase in several proteins involved in axon-glia interaction, such as MAG, N-CAM, and F3/Contactin. This co-culture system represents a potentially powerful tool to study neuron-glia interactions that occur during myelinogenesis and the role of Tf in this process.
Paez
Pablo M
PM
Instituto de Química y Fisicoquímica Biológica (IQUIFIB), UBA-CONICET, and Departamento de Química Biológica, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Buenos Aires, Argentina.
García
Corina I
CI
Campagnoni
Anthony T
AT
Soto
Eduardo F
EF
Pasquini
Juana M
JM
D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D015816:Cell Adhesion Molecules, Neuronal / Q000378:metabolism; D002450:Cell Communication / Q000235:genetics; D002454:Cell Differentiation / Q000235:genetics*; D002460:Cell Line; D002490:Central Nervous System / Q000166:cytology / Q000254:growth & development / Q000378:metabolism; D002999:Clone Cells / Q000166:cytology / Q000378:metabolism; D018920:Coculture Techniques / Q000379:methods; D058966:Contactins; D018507:Gene Expression Regulation, Developmental / Q000235:genetics; D006801:Humans; D051379:Mice; D004676:Myelin Basic Protein / Q000378:metabolism; D009186:Myelin Sheath / Q000378:metabolism / Q000648:ultrastructure; D018992:Myelin-Associated Glycoprotein / Q000378:metabolism; D019006:Neural Cell Adhesion Molecules / Q000378:metabolism; D009474:Neurons / Q000378:metabolism; D009836:Oligodendroglia / Q000166:cytology / Q000378:metabolism*; D051381:Rats; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism; D014162:Transfection / Q000379:methods; D014168:Transferrin / Q000235:genetics* / Q000378:metabolism*
D015415:Biomarkers; D015816:Cell Adhesion Molecules, Neuronal; D058966:Contactins; D004676:Myelin Basic Protein; D018992:Myelin-Associated Glycoprotein; D019006:Neural Cell Adhesion Molecules; D014168:Transferrin