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J. Biol. Chem., Vol. 282, Issue 12, 9172-9181, March 23, 2007

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Neural Stem/Progenitor Cells Express 20 Tenascin C Isoforms That Are Differentially Regulated by Pax6^*

Alexander von Holst¹², Ursula Egbers¹, Alain Prochiantz, and Andreas Faissner³

From the Department of Cell Morphology and Molecular Neurobiology, Ruhr-University Bochum, D-44780 Bochum, Germany and CNRS UMR 8542, Development and Neuropharmacology, Ecole Normale Superieure, 46 rue d'Ulm, 75005 Paris, Cedex 05, France

Tenascin C (Tnc) is an alternatively spliced, multimodular extracellular matrix glycoprotein present in the ventricular zone of the developing brain. Pax6-deficient small eye (sey) mouse mutants show an altered Tnc expression pattern. Here, we investigated the expression of Tnc isoforms in neural stem/progenitor cells and their regulation by the paired-box transcription factor Pax6. Neural stem/progenitor cells cultured as neurospheres strongly expressed Tnc on the protein level. The Tnc isoform expression in neural stem/progenitor cells was analyzed by reverse transcriptase-PCR and dot blot Southern hybridization. In total, 20 different Tnc isoforms were detected in neurospheres derived from embryonic fore-brain cell suspensions. The Tnc isoform containing the fibronectin type III domains A1A4BD is novel and might be neural stem/progenitor cell-specific. Transient overexpression of Pax6 in neurospheres of the medial ganglionic eminence did not alter the total Tnc mRNA expression level but showed a pronounced regulative effect on different Tnc isoforms. The larger Tnc isoforms containing four, five, and six additional alternatively spliced fibronectin type III domains were up-regulated, whereas the small Tnc isoforms without any or with one additional domain were down-regulated. Thus, Pax6 is a homeodomain protein that also modulates the splicing machinery. We conclude that the combinatorial code of Tnc isoform expression in the neural stem/progenitor cell is complex and regulated by Pax6. These findings suggest a functional significance for individual Tnc isoforms in neural stem/progenitor cells.

Received for publication, August 22, 2006 , and in revised form, January 8, 2007.

^* This work was supported by Deutsche Forschungsgemeinschaft graduate program GRK 736 (to U. E.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ Both authors contributed equally.

² To whom correspondence may be addressed: Universitätsstr. 150, NDEF 05/339, D-44780 Bochum, Germany. Tel.: 49-234-3225812; Fax: 49-234-3214313; E-mail: Alexander.vonHolst{at}ruhr-uni-bochum.de.

³ To whom correspondence may be addressed: Universitätsstr. 150, NDEF 05/594, D-44780 Bochum, Germany. Tel.: 49-234-3223851; Fax: 49-234-3214313; E-mail: Andreas.Faissner{at}ruhr-uni-bochum.de.

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