Expression of neuronal markers during NTera2/cloneD1 differentiation by cell aggregation method.
Human teratocarcinoma NTera2/cloneD1 (NT2) cells are able to generate postmitotic neurons in response to retinoic acid (RA) and for this reason these cells provide an important tool to study human neurogenesis in vitro. We have obtained neurons by treating NT2 aggregated cells with RA for solely 14 days. RT-PCR assays showed that NT2 cells express mRNAs of several neural bHLH genes such as Hes1, Ngn1, Mash1, NeuroD, Math1 and Pax6, just in the early days of RA exposure. In particular, we reported for the first time that RA treatment was followed by a modulation of endogenous Ngn1 and Math1 transcripts. RT-PCR and Western blotting experiments also demonstrated expression of typical neuronal markers such as GluR, MAP2, Tau and NeuN. Knowledge of the expression pattern of the different neuronal genes during NT2 commitment could be used to investigate alterations in the molecular pathways involved in the human neuronal differentiation.
Megiorni
Francesca
F
Department of Experimental Medicine and Pathology, La Sapienza University, Viale Regina Elena 324, 00161 Rome, Italy.
Mora
Barbara
B
Indovina
Paola
P
Mazzilli
Maria Cristina
MC
D000970:Antineoplastic Agents / Q000494:pharmacology; D015415:Biomarkers / Q000032:analysis*; D015153:Blotting, Western; D002449:Cell Aggregation; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D045744:Cell Line, Tumor; D002999:Clone Cells; D017931:DNA Primers; D018257:Helix-Loop-Helix Motifs / Q000187:drug effects / Q000502:physiology; D006801:Humans; D009419:Nerve Tissue Proteins / Q000096:biosynthesis* / Q000187:drug effects; D009474:Neurons / Q000166:cytology* / Q000502:physiology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018243:Teratocarcinoma; D014157:Transcription Factors / Q000096:biosynthesis* / Q000187:drug effects; D014212:Tretinoin / Q000494:pharmacology
D000970:Antineoplastic Agents; D015415:Biomarkers; D017931:DNA Primers; D009419:Nerve Tissue Proteins; D014157:Transcription Factors; D014212:Tretinoin