Retrograde axonal transport of poliovirus and EV71 in motor neurons

Figures

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  Fig. 1. PV RNA genome is transported retrogradely with capsid in MNs expressing hPVR (A) Double-labeled PV was added to the axonal compartment followed by observation under fluorescent microscope. AlexaFluor488-labeled capsid and SYTO82-labeled RNA genome are in green and red, respectively. Green and blue circles indicate the relative position of two moving axonal organelles. The time in second after starting the observation is shown. The brightness of the left of the images was enhanced. Scale bar, 10 µm. (B) Kymographs for the organelle marked by the blue circles. Scale bar, 1 µm. (C to J) After the addition of PV to either the somatic or axonal compartment of MNs cultured in a microfluidic device, the medium was collected at the indicated time points followed by titration. Pink and green indicate titer in somatic and axonal compartments, respectively. Error bars indicate SD. (C, D) Somatic compartment infection of hPVR-Tg MNs. (E, F) Axonal compartment infection of hPVR-Tg MNs. (G, H) Somatic compartment infection of non-Tg MNs. (I, J) Axonal compartment infection of non-Tg MNs. (D, F, H, and J) Representative images of somatic compartment at 6 h.a.i. Scale bars, 5 µm. (C, D, G to J)…

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  Fig. 2. EV71 is retrogradely transported through the sciatic nerve in hSCARB2-Tg mice (A) Number of non-paralysis time course after intramuscular injection with EV71 (n = 3). Repeated experiments showed the same trend. (B) Virus titer in spinal cord or muscle (left or right) 72 h after intramuscular injection with EV71. Filled circle, square, and triangle indicate spinal cord, muscle (left), and muscle (right), respectively. Number shown under horizontal axis indicates the mouse identification number. Mouse 4 and 5 showed paralysis at 72 h.a.i., indicated as P in the graph. (C) Viral titer in spinal cord or muscle (left or right) at 72 h.a.i from experiment shown in B. Each circle indicates one mouse. (D) Viral titer in spinal cord 72 h after intramuscular injection with EV71. Prior to the viral injection, sciatic nerve transection or sham operation were carried out. Each circle indicates one mouse. Median and range are shown in C and mean and range are shown in D.

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  Fig. 3. EV71 is retrogradely transported in MNs expressing hSCARB2 After addition of EV71 in either the somatic or axonal compartment of MNs cultured in a microfluidic device, the medium was collected at indicated time points followed by titration. (A) Somatic compartment infection of hSCARB2-Tg MNs. (C) Axonal compartment infection of hSCARB2-Tg MNs. (B, D) Representative immunofluorescence images of somatic compartment at 8 h.a.i. Infection of non-Tg MNs in the somatic (E) and axonal (F) compartment. Scale bars, 5 µm.

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  Fig. 4. hSCARB2 is retrogradely transported in MNs expressing hSCARB2 hSCARB2-Tg MNs were infected with EV71 in the presence of control IgG (A, C) or anti-hSCARB2 antibodies (B, D) added to the somatic (A, B) or axonal (C, D) compartment. Cells were fixed 16 h.a.i. followed by immunofluorescence. Representative images show DAPI in blue, control and anti-hSCARB2 antibodies in green, whereas EV71 is in red. Scale bars, 5 µm. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)