Figure 6

Monitoring intra-cellular signaling activity in the adult brain.

(a–b3) Electroporation of a Wnt reporter vector into the lateral ventricle. RFP expression vector with CAG promoter was co-transfected with pTOP-dGFP that contains Lef/Tcf response elements (a). At 24 hours after electroporation, GFP expression was detected in ependymal cells by electroporation of pTOP-dGFP (b1–b3). Only a background GFP expression was detected by pFOP-dGFP that contains mutated Lef/Tcf response elements (d1–d3). (e–f3) Electroporation of pHes5-GFPd2, in which GFP tagged with de-stabilizing signal (e). After the focal electroporation, GFP expression was detected in ependymal cells (f1–f3). (g–i3) To monitor canonical Notch signaling activity, p12xCSL-GFP vector that expresses GFP under the control of CSL (CBF)-response elements, was electroporated to the SVZ (g). GFP expression was detected in the SVZ and striatum (arrowheads in h1–h3). An astrocytic cell that was positive for GFP (i1–i3). A background GFP expression was detected by electroporation of pCBFRE(mt)-GFP vector that contains mutated CBF response elements (j–k3). Scale bars: 25 μm in (b1), (d1), (f1), (i1) and (k1); 100 μm (h1).