Abstract
Systemic injection of a thymidine analogue such as bromodeoxyuridine (BrdU) in vertebrates is commonly used to detect and study cell production during development, adulthood, and pathology, particularly in studies of adult neurogenesis. Although researchers are applying this technique to multiple species in various physiological conditions, the rate of BrdU clearance from the serum remains unknown in most cases. Changes in this clearance rate as a function of the species, sex or endocrine condition could however profoundly affect the interpretation of the results. We describe a rapid, sensitive, but simple bioassay for post-injection detection and quantification of BrdU in serum. This procedure was shown to be suitable for determining the length of time a thymidine analogue remains in the bloodstream of one avian species and seems applicable to any vertebrate provided sufficiently large blood samples can be collected. This technique was used to demonstrate that, in canaries, BrdU injected at a dose of 100 mg/kg is no longer available for incorporation into DNA between 30 and 60 min post-injection, a delay shorter than anticipated based on the available literature. Preliminary data suggest a similar fast clearance in Japanese quail and mice.
Highlights
The injection of thymidine analogues such as bromodeoxyuridine (BrdU) that are incorporated into DNA during its replication continues to be the best and most commonly used technique for investigating proliferating populations of cells in vivo [1], [2]
Since newly-formed neuronal precursors very often do not undergo many subsequent divisions, the incorporated BrdU can be used to trace the survival of the new neurons (e.g., [7,8,9,10,11])
Multiple studies have for example identified differences in the number of new neurons labelled with BrdU as a function of the sex or endocrine conditions of the subjects
Summary
The injection of thymidine analogues such as bromodeoxyuridine (BrdU) that are incorporated into DNA during its replication continues to be the best and most commonly used technique for investigating proliferating populations of cells in vivo [1], [2]. Multiple studies have for example identified differences in the number of new neurons labelled with BrdU as a function of the sex or endocrine conditions of the subjects (for recent reviews see [17,18,19]). This could reflect true differences in the rate of neurogenesis, but undetected changes in BrdU clearance from the serum could have the same effect
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