The expression level of the orphan nuclear receptor GCNF (germ cell nuclear factor) is critical for neuronal differentiation.
The germ cell nuclear factor (GCNF) is essential for normal embryonic development and gametogenesis. To test the prediction that GCNF is additionally required for neuronal differentiation, we used the mouse embryonal carcinoma cell line PCC7-Mz1, which represents an advantageous model to study neuronal cells from the stage of fate choice until the acquirement of functional competence. We generated stable transfectants that express gcnf sense or antisense RNA under the control of a tetracycline-regulated promoter. After retinoic acid-induced withdrawal from the cell cycle, sense clones developed a neuron network with changed properties, and the time course of neuron maturation was shortened. Consistent with these data, differentiation of neuronal precursor cells was impaired in antisense cultures. This involved a delay in 1) the down-regulation of nestin, a marker for undifferentiated neuroepithelial cells and stem cells of the central nervous system, and 2) up-regulation of the somatodendritic protein microtubule-associated protein 2 and the synaptic vesicle protein synaptophysin. Neuronal cells in the antisense cultures acquired functional competence, although with a significant delay. Our data propose that the level of GCNF is critical for differentiation and maturation of neuronal precursor cells.
Institute of Physiological Biochemistry and Pathobiochemistry, Johannes Gutenberg University, Medical School, Duesberg Weg 6, 55099 Mainz, Germany.
Ulrike
U
Sattler
Marek
M
Samochocki
Alfred
A
Maelicke
Christina
C
Zechel
D000818:Animals; D002453:Cell Cycle / Q000187:drug effects; D002454:Cell Differentiation / Q000235:genetics; D016764:Cell Polarity; D004268:DNA-Binding Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D015536:Down-Regulation; D019922:GAP-43 Protein / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D015870:Gene Expression; D007381:Intermediate Filament Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D064231:Nestin; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D057136:Nuclear Receptor Subfamily 6, Group A, Member 1; D018408:Patch-Clamp Techniques; D016372:RNA, Antisense / Q000235:genetics; D018160:Receptors, Cytoplasmic and Nuclear / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D016708:Synaptophysin / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D014212:Tretinoin / Q000494:pharmacology; D015854:Up-Regulation
D004268:DNA-Binding Proteins; D019922:GAP-43 Protein; D007381:Intermediate Filament Proteins; D008869:Microtubule-Associated Proteins; D009419:Nerve Tissue Proteins; C577485:Nes protein, mouse; D064231:Nestin; C539954:Nr6a1 protein, mouse; D057136:Nuclear Receptor Subfamily 6, Group A, Member 1; D016372:RNA, Antisense; D018160:Receptors, Cytoplasmic and Nuclear; D016708:Synaptophysin; D014212:Tretinoin