Figure 4

Cell-to-cell protein transfer is specific to Tau protein WT. (a) eGFP is not transported in secondary connected neurons. LVs encoding eGFP were bilaterally injected into the CA1 layer of rat brains (IS bregma -5.3, n = 3). Eight months later, the animals were sacrificed, and brains sections processed for immunofluorescence assays to detect eGFP. Sections from the prelimbic (bregma +4.7), external capsule (bregma -1.8), CA1 (IS, bregma -5.3) and ectorhinal cortex (bregma -7.8) are shown. The scale bars are indicated on the figure. (b) Restricted lentiviral insertion in the vicinity of the IS. PBS (left panel, n = 3) or LVs encoding hTau46^WT (middle panel, n = 3) or hTau46^P301L (right panel, n = 3) were bilaterally injected into the CA1 layer of rat brains. One month later, the brains were dissected, and coronal sections of 1 mm thickness (indicated as the position relative to bregma) were prepared using an acrylic rat brain matrix. Total RNA was extracted from these sections to generate cDNA using RT-PCR. cDNAs were amplified using oligonucleotides specific to human Tau (hTau, 117 base pairs) or murine Tau (70 base pairs). The positive control was prepared by amplifying the plasmid containing hTau sequence. The lower parts represent the mean +/-SEM of the relative density (hTau/mTau) coming from the three rats. These data showed that Tau transport is a specific mechanism since neither eGFP nor viral genome is found associated to distant secondary brain areas.