Skip to main content
Fig. 4 | Molecular Neurodegeneration

Fig. 4

From: A novel H129-based anterograde monosynaptic tracer exhibits features of strong labeling intensity, high tracing efficiency, and reduced retrograde labeling

Fig. 4

Significantly reduced retrograde labeling of gKmut pseudotyped H129-dgK-G4. A, B. In vitro comparison of retrograde labeling by axon terminal invasion between H129-dgK-G4(gKwt) and H129-dgK-G4(gKmut). 1 × 106 fetal mouse cortical neurons were seeded into the soma chamber of the microfluidic plates, and cultured for 14 days to allow axons to grow through the microchannel and reach the contralateral chamber. Then, 1 × 106 pfu of H129-dgK-G4(gKwt) or H129-dgK-G4(gKmut) was added to the axon terminal side. GFP-labeled neurons were monitored and counted at 1 dpi. Shown are the representative images from 6 microchannel plates of each group performed in 2 independent experiments (A). The GFP positive neurons were counted in each chamber, and the statistical significance was analyzed by Student’s t-test. ***, p < 0.001 (B). C, D. In vivo comparison of retrogradely labeling between H129-dgK-G4(gKwt) and H129-dgK-G4(gKmut). H129-dgK-G4(gKwt) or H129-dgK-G4(gKmut) was injected into CA1 (indicated with the solid-line box) of wildtype C57BL/6 mice with the same dosage (5.0 × 108 pfu/ml, 100 nl), and the brains were collected at 1 dpi. The numbers of the GFP-labeled neurons at the ectorhinal cortex (Ect), an upstream nucleus of CA1, were examined and counted in each brain. Shown are the representative images from 3 mice of each group (C, upper panel), and the magnified images of Ect (indicated with the dashed-line box) (C, bottom panel). The left hemisphere displays the corresponding sections from Allen Brain Reference Atlases (Image credit: Allen Institute). Numbers of the GFP+ neuron in the position-matched brain slices were counted (11 slices/mouse × 3 mice), and statistical significance was analyzed by LME. **, p < 0.01 (D)

Back to article page