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As we were most interested in performing our studies in mouse neurons, we established the efficacy of reagents in HEK293T cells expressing transduced mThap1.
To quantify the differences in subcellular morphology between human and mouse neurons, we compared the segment length of both apical and basal dendrites using linear and quadratic polynomials in a MLM (Hox 2010).
In titrating GSH levels by inhibiting glutathione cysteine ligase in cultured mouse neurons, we also observed that GSH depletion is more important than ROS elevation in causing increased neruon death in aging and AD neurons (Ghosh et al., 2014).
However, as the splice variant identified by Giudici et al. seems to be limited to mouse neurons, we propose that the much more ubiquitous human PIPKIγ_i5 could serve to perform similar biological functions in non-neuronal cells.
After transfection into either HEK293 cells or primary E14.5 cortical mouse neurons, we observed a mean of ~3.5-fold decrease in luciferase expression with the high-risk alleles (DNR > 11, i.e., 12, 13, or 14 repeats) compared with the low risk allele (DNR = 11 repeats) (P < 0.0001).
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Using two-photon calcium imaging of mouse hippocampal neurons we find that co-time-tuned neurons exhibit enhanced spontaneous activity correlations that increase just prior to learning.
In mouse cortical neurons, we observed spatacsin expression in axonal (TAU) and dendritic (MAP2) processes (Fig. 2A) with no preference of spatacsin localization to either compartment (Fig. 2B).
In siSPG11+ mouse cortical neurons, we observed a marked reduction of neurite complexity at Day 2 compared with MOCK and siLuc transfected neurons (Fig. 4A, part i, B and D H), paralleling our findings in SPG11 patient-derived neurons.
Therefore, in this in vitro study using cultured mouse cortical neurons, we have investigated axon degeneration following excitotoxicity induced by kainic acid to determine mechanistic similarities to other forms of axon degeneration.
By employing a complementary approach using human and mouse cortical neurons, we provide the first evidence that spatacsin has a major impact on neurite plasticity through maintaining cytoskeleton stability and SV transport.
Third, we made neuronal-specific transgenic mice and found that overexpressing the miR-23b and miR-27b in mouse neurons inhibited the neuronal apoptosis induced by intrauterine hypoxia.
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Since I tried Ludwig back in 2017, I have been constantly using it in both editing and translation. Ever since, I suggest it to my translators at ProSciEditing.

Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com