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Therefore, the physiological and quantitative analysis of neuronal subpopulations at different ages is critical to studies of neurogenesis.
Such approaches allow cells of different ages to be identified by labeling them according to their probable date of birth.
Naturally there some other minor issues are involved in its use, though these have largely been solved over the years.
One of the most interesting parameters that can be assessed by the labeling of dividing cells is survival time, defined as the time between the incorporation of the Brd U and that of the animal’s sacrifice.
After the animal is sacrificed, staining is detected by immunohistochemistry using antibodies specifically directed against the analog.
Brd U has been the marker of choice in recent years for several reasons, in part because this method requires no radioactivity unlike the use of tritiated thymidine, which for decades was used to label dividing cell populations during brain development.
This review summarizes the current research carried out using these techniques and outlines some of the key applications.
The acclaimed working hypothesis that different subpopulations of newborn, differentiating neurons could be playing different roles arouses great interest.The age of labeled cells is equivalent to the survival time (i.e., cells are 1 month old if the animal injected with Brd U was sacrificed 1 month after Brd U injection).A key limitation of this method is its ability to recognize only a single pool of Brd U incorporated into the body, regardless of when and how it was administered.Because the marker is persistent, it is possible to retrospectively conclude with confidence that a marked cell must have undergone cell division at the time when the marker was injected (sic)” (Kempermann, 2006).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).