By flow-sorting immature Crx-expressing cells from various developmental time points and transplanting the genetically labelled populations into wild-type or degenerating mouse retinae, we were able to compare their integration competency and ability to generate cones after transplantation.
The idea is to be able to elicit a "truthful" response on sensitive feature(s) from the sampled respondents (of a finite labeled population of respondents), so that eventually the population mean of the sensitive feature can be unbiasedly estimated.
Antibody treated and untreated KO mice were injected with BrdU for 4 days, then subsequently CD8+ T cells response in the BrdU labeled population was measured at day 6 and 14 p.i. (Figure 3A).
Because neurites are packed so densely in neuropil (∼10 40 neurites per voxel of a typical diffraction limited microscope), for a densely labeled population of neurons the fluorescence from all such neurites tends to blend together, making individual neurites indiscernible.
The mouse studies reported thus far have not applied genetic labeling technologies to the morphologic analysis of neurons with large and complex arbors, an application that requires extremely sparse labeling and that is only rendered practical by limiting the labeled population to one or a few predetermined neuronal subtypes.
In mammals, genetically-directed cell labeling technologies have not yet been applied to the morphologic analysis of neurons with very large and complex arbors, an application that requires extremely sparse labeling and that is only rendered practical by limiting the labeled population to one or a few predetermined neuronal subtypes.
2) Figure 2: The clonal nature of the labeled population is crucial for the interpretation of the data.
Each labeled population of NIH3T3 were assigned specific treatments, for example, light, control; medium, TNF α/CHX; and heavy, TNF α/CHX+Y27632.
Each labeled population was used in each treatment condition, with Blebbistatin substituting for Y27632 in one set of experiments, for a total of six replicates.
Results indicated nearly a 40% increase in type II-like SGN fibers (among the labeled population) in Nrp2 +/− cochleae at P1, confirming the phenotype observed in embryonic cochleae from Nrp2 mutants.
