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Second, Ohata et al. analyzed motor neuron number and patterning by in situ for islet1 and islet2 and concluded that motor neuron number and patterning are unaltered in twd.
Because primary motor neuron number is increased in srn, it is difficult to separate direct effects of Notch-Delta signaling on presynaptic differentiation from indirect effects on neurogenesis.
While the number of motor neurons was normal in Cra1/+ mice at three months of age, a severe reduction in motor neuron number was reported at 16 months of age in Cra1/+ mice and 18 months of age in Loa/+ mice, consistent with the progressive symptoms.
For quantification of motor neuron number, CC1+ oligodendrocyte number, casp3+ cell number and GFAP or MBP levels, at least three sections/animal, three animals/group were used.
For immunohistochemical analysis of motor neuron number, sections were first rehydrated 40 min in PBS followed by 10 min permeabilization in 0.1% Triton X.
The first analysis, from Reaume et al. (1996), found no reduction in motor neuron number at 4 months of age, but an increase in small neurons and astrocytes in spinal cord ventral horns.
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Electromyography recordings, motor neurons counts, and neuromuscular junction analysis in aged (18 22 months) Loa/+ and Cra1/+ mice all demonstrate normal motor neuron numbers and innervation of muscle [11], [12].
Loss of motor neurons by TDP-43WT, TDP-43A315T and TDP-43ΔNLS was further preceded by transient increase in Caspase-3+ motor neuron numbers, mirroring the relative impact of TDP-43 variants on overall reduction of motor neurons numbers (Fig. 5C).
However, in contrast to TDP-43WT and similar to TDP-43FFLL and TDP-43CTF, TDP-43NTF expression had no significant impacts on maintenance of normal motor neuron numbers (Fig. 5B), nor on motor neuron apoptosis levels (Fig. 5C).
However, for both TDP-43CTF and TDP-43FFLL,ΔNLS we observed a transient mild increase in Casp-3+ motor neurons at 2 days after transfection (Fig. 5C); which, however, was apparently insufficient to result in measurable reduction of motor neuron numbers at later stages (Fig. 5B).
Even when paralysis ensues, motor neuron numbers remain unaffected.
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