Therefore, the injection rate in conjunction with the survival time can produce large differences, and in the worst cases, cell populations that are markedly heterogeneous in terms of age may be labeled equally.
Within the framework of the experimental design, the investigator must judge what heterogeneity in terms of age can be accepted in the target cell population.
The cells born in the adult dentate gyrus are derived from neural stem cells (NSC; type-1 cells); these cells are either quiescent (a small proportion) or divide slowly, to generate another group of rapidly dividing cells known as intermediate progenitor cells (type-2a cells).
The protocol involves the administration of the thymidine analog (normally by intraperitoneal injection, and to a lesser extent via drinking water) which incorporates into the dual helix of any cell actively synthesizing DNA during the period the product remains systemically available and active in the body (usually around 2 h or less).
This issue is particularly important in short experiments (i.e., days).
If the animal is injected with Brd U over n days, and sacrificed 1 week after the last injection, the cohort of labeled cells is considered to be between 7 and 7−n days old.
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This has significant implications: all incorporated Brd U is detected as a single signal, and hence, the cell populations that have incorporated Brd U are indistinguishable.