Antisense morpholino oligonucleotides (MOs) block NF180 expression in RNs.
A, Design of the MO, the antisense sequence (in red) was complementary to the region surrounding the start codon (-4 to +21) of the NF180 gene. B, Fluorescently-tagged MOs were delivered to spinal-projecting neurons by retrograde transport from a spinal cord TX. After 2 or 4 wks, the brains were processed for immuno-histochemistry with LCM16, a mAb specific for NF180. Intensity of labeling was reduced in antisense MO-treated brains compared to brains treated with control MOs. Insets are fluorescent images of the same brains showing the presence of MOs. C, Lampreys were treated as in B, and the proteins extracted from the supernatant (Super) and pellet of homogenized, centrifuged brain and spinal cord rostral to the transection. Antisense MOs (A) significantly reduced the expression of NF180 in Super and pellet fractions of both 2- and 4-week groups compared with control (C). There was less effect on the pellet and no apparent effect on another neuronal intermediate filament, nIF50.