Incorporation of Thymidine and its Analogue, Bromodeoxyuridine, into Embryos and Maternal Tissues of the Mouse

Packard, David S.; Menzies, Robert A.; Skalko, Richard G.

doi:10.1111/j.1432-0436.1973.tb00137.x

Cited by 151 publications

(83 citation statements)

References 26 publications

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“…This extended labeling time explains previous findings that the number of [ 3 H]thymidinelabeled cells in the adult dentate gyrus doubles between 2 and 24 hours after injection (Okano et al, 1996) but more than doubles between 1 hour and 24 hours (Cameron et al, 1993); the 1.9-hour labeling time means that additional cells are labeled after the 1-hour survival time, but not after 2 hours. Similar evidence from the current study, that the BrdU-labeled cell number at 24 hours is twice the number at 2 hours (experiment 2), indicates that BrdU also stops labeling cells in the adult brain within 2 hours, in agreement with studies showing that BrdU clearance time is identical to that of thymidine (Packard et al, 1973;Hayes and Nowakowski, 2000).…”

Section: Experiments 4: Cell Cycle Time Calculationsupporting

confidence: 79%

“…It has long been believed that [ 3 H]thymidine and BrdU are cleared from the body, and presumably the brain, very rapidly, i.e., within 30 minutes (Packard et al, 1973); consequently an injection of one of these markers would act as a "pulse," labeling a discrete cohort of S-phase cells. However, a recent embryonic study suggests that, in fact, both markers may continue to label cells in the brain for a considerably longer time, 5-6 hours (Hayes and Nowa- N bt ).…”

Section: Experiments 4: Cell Cycle Time Calculationmentioning

confidence: 99%

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Adult neurogenesis produces a large pool of new granule cells in the dentate gyrus

Cameron

McKay

2001

J of Comparative Neurology

1,411

1,137

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Knowing the rate of addition of new granule cells to the adult dentate gyrus is critical to understanding the function of adult neurogenesis. Despite the large number of studies of neurogenesis in the adult dentate gyrus, basic questions about the magnitude of this phenomenon have never been addressed. The S-phase marker bromodeoxyuridine (BrdU) has been extensively used in recent studies of adult neurogenesis, but it has been carefully tested only in the embryonic brain. Here, we show that a high dose of BrdU (300 mg/kg) is a specific, quantitative, and nontoxic marker of dividing cells in the adult rat dentate gyrus, whereas lower doses label only a fraction of the S-phase cells. By using this high dose of BrdU along with a second S-phase marker, [ 3 H]thymidine, we found that young adult rats have 9,400 dividing cells proliferating with a cell cycle time of 25 hours, which would generate 9,000 new cells each day, or more than 250,000 per month. Within 5-12 days of BrdU injection, a substantial pool of immature granule neurons, 50% of all BrdU-labeled cells in the dentate gyrus, could be identified with neuron-specific antibodies TuJ1 and TUC-4. This number of new granule neurons generated each month is 6% of the total size of the granule cell population and 30 -60% of the size of the afferent and efferent populations (West et al. [1991] Anat Rec 231:482-497; Mulders et al. [1997] J Comp Neurol 385:83-94). The large number of the adult-generated granule cells supports the idea that these new neurons play an important role in hippocampal function.

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Section: Experiments 4: Cell Cycle Time Calculationsupporting

confidence: 79%

Section: Experiments 4: Cell Cycle Time Calculationmentioning

confidence: 99%

Adult neurogenesis produces a large pool of new granule cells in the dentate gyrus

Cameron

McKay

2001

J of Comparative Neurology

1,411

1,137

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“…Because the systemically administered BrdU is quickly cleared from the body within 1 hr (Packard et al, 1973;Cameron and McKay, 2001), this experiment allows us to "pulse label" the hippocampal neurons that were synthesizing DNA on the second day after stroke and "chase" the outcomes of these cells at intervals of 5 and 26 d, respectively. We found that at a 5 d interval, the CA1-hippocampal subfield remained densely populated by pyknotic and BrdU-labeled nuclei (Fig.…”

Section: Brdu-incorporated Neurons Only Temporarily Exist In the Hippmentioning

confidence: 99%

Hypoxia-Ischemia Induces DNA Synthesis without Cell Proliferation in Dying Neurons in Adult Rodent Brain

et al. 2004

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Recent studies suggest that postmitotic neurons can reenter the cell cycle as a prelude to apoptosis after brain injury. However, most dying neurons do not pass the G 1 /S-phase checkpoint to resume DNA synthesis. The specific factors that trigger abortive DNA synthesis are not characterized. Here we show that the combination of hypoxia and ischemia induces adult rodent neurons to resume DNA synthesis as indicated by incorporation of bromodeoxyuridine (BrdU) and expression of G 1 /S-phase cell cycle transition markers. After hypoxia-ischemia, the majority of BrdU-and neuronal nuclei (NeuN)-immunoreactive cells are also terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end labeling (TUNEL)-stained, suggesting that they undergo apoptosis. BrdU ϩ neurons, labeled shortly after hypoxia-ischemia, persist for Ͼ5 d but eventually disappear by 28 d. Before disappearing, these BrdU ϩ /NeuN ϩ / TUNEL ϩ neurons express the proliferating cell marker Ki67, lose the G 1 -phase cyclin-dependent kinase (CDK) inhibitors p16INK4 and p27Kip1 and show induction of the late G 1 /S-phase CDK2 activity and phosphorylation of the retinoblastoma protein. This contrasts to kainic acid excitotoxicity and traumatic brain injury, which produce TUNEL-positive neurons without evidence of DNA synthesis or G 1 /S-phase cell cycle transition. These findings suggest that hypoxia-ischemia triggers neurons to reenter the cell cycle and resume apoptosis-associated DNA synthesis in brain. Our data also suggest that the demonstration of neurogenesis after brain injury requires not only BrdU uptake and mature neuronal markers but also evidence showing absence of apoptotic markers. Manipulating the aberrant apoptosis-associated DNA synthesis that occurs with hypoxia-ischemia and perhaps neurodegenerative diseases could promote neuronal survival and neurogenesis.

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“…To investigate if Pact Ϫ/Ϫ hypoplasia was due to decreased cell proliferation in the knockout animals, we measured the levels of Ki67 and incorporation of bromodeoxyuridine (BrdU), markers for proliferating cells. Ki67 is expressed in actively dividing cells, but not quiescent cells (28), whereas BrdU is incorporated into new DNA during the S-phase of the cell cycle (29). Western blotting of whole pituitary extracts revealed decreased Ki67 levels in P21 and P90 Pact Ϫ/Ϫ mice compared to Wt littermates (Fig.…”

Section: Pact Is Essential For Postnatal Development Of the Anterior mentioning

confidence: 99%

The double-stranded RNA-binding protein, PACT, is required for postnatal anterior pituitary proliferation

Peters

Seachrist

Keri

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

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Incorporation of Thymidine and its Analogue, Bromodeoxyuridine, into Embryos and Maternal Tissues of the Mouse

Cited by 151 publications

References 26 publications

Adult neurogenesis produces a large pool of new granule cells in the dentate gyrus

Adult neurogenesis produces a large pool of new granule cells in the dentate gyrus

Hypoxia-Ischemia Induces DNA Synthesis without Cell Proliferation in Dying Neurons in Adult Rodent Brain

The double-stranded RNA-binding protein, PACT, is required for postnatal anterior pituitary proliferation

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