The utility of Ki-67 and BrdU as proliferative markers of adult neurogenesis

Kee, Nohjin; Sivalingam, Sugirthan; Boonstra, Rudy; Wojtowicz, J. Martin

doi:10.1016/s0165-0270(02)00007-9

Cited by 682 publications

(541 citation statements)

References 30 publications

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“…To have a second confirmation of the presence of proliferating cells in the cultures, we stained parallel samples for Ki-67 (a marker expressed during most of the cell cycle 26 ) and, as expected, we found more cells still in cycle in Bcl-X L cultures, from day 5 onwards (Figure 5d and e).…”

Section: Resultssupporting

confidence: 73%

Bcl-XL modulates the differentiation of immortalized human neural stem cells

et al. 2007

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Understanding basic processes of human neural stem cell (hNSC) biology and differentiation is crucial for the development of cell replacement therapies. Bcl-X L has been reported to enhance dopaminergic neuron generation from hNSCs and mouse embryonic stem cells. In this work, we wanted to study, at the cellular level, the effects that Bcl-X L may exert on cell death during differentiation of hNSCs, and also on cell fate decisions and differentiation. To this end, we have used both v-myc immortalized (hNS1 cell line) and non-immortalized neurosphere cultures of hNSCs. In culture, using different experimental settings, we have consistently found that Bcl-X L enhances neuron generation while precluding glia generation. These effects do not arise from a glia-to-neuron shift (changes in fate decisions taken by precursors) or by only cell death counteraction, but, rather, data point to Bcl-X L increasing proliferation of neuronal progenitors, and inhibiting the differentiation of glial precursors. In vivo, after transplantation into the aged rat striatum, Bcl-X L overexpressing hNS1 cells generated more neurons and less glia than the control ones, confirming the results obtained in vitro. These results indicate an action of Bcl-X L modulating hNSCs differentiation, and may be thus important for the future development of cell therapy strategies for the diseased mammalian brain. The efficient generation of human neurons and glia from stem cells is the object of intense investigation, in the context of basic and preclinical cell replacement research. 1,2 Different means of genetic and epigenetic manipulations of stem cell cultures are being tested, aiming at enhancing their ability to generate the desired cell types. [3][4][5][6] In previous studies, we and others have demonstrated that Bcl-X L overexpression has a major impact on the generation of dopaminergic neurons from human neural stem cells (hNSCs) and mouse embryonic stem cells (mES). 7,8 Also, recent data from conditional Bcl-X L mutant mice add support to these observations. 9,10 In the present study, we are extending these observations and analyzing in detail the effects of Bcl-X L on hNSCs differentiation, focusing on precursor cell fate choices, cell death, and precursor proliferation.Bcl-X L is the most potent antiapoptotic protein among Bcl-2 family members, both in vitro and in vivo. 11-14 More specifically, Bcl-X L is essential for neuronal survival during brain development and in the adult central nervous system (CNS) (knockout mice and gene-expression studies 15,16 ). In addition to its antiapoptotic role, recent studies have described new roles of Bcl-X L in cell physiology -effects on Ca 2 þ homeostasis and gene expression 17 and synaptic transmission regulation 18 -under not necessarily apoptotic conditions. Several other studies have also described the role of Bcl-X L in the control of cell cycle, 19,20 a process well known to be tightly linked to progression of precursor cells toward neuronal and glia differentiation. 21,22 In this work, we aimed...

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Section: Resultssupporting

confidence: 73%

Bcl-XL modulates the differentiation of immortalized human neural stem cells

et al. 2007

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“…34,47 As previously described, 10 more than 90% of cells labeled with BrdU or Ki67 antibodies were found tightly associated in discrete clusters along the SGZ with a small proportion present as isolated cells. Few labeled cells were located in the granular and molecular layers (data not shown).…”

Section: Neurogenesis In the Adult Dg Is Influenced By Thyroid Hormonmentioning

confidence: 63%

“…This agrees with the finding of more than 90% of the labeled cells in proliferative clusters, and the similarity in the distribution of cells expressing Ki67, which labels proliferating cells in any phase of the cell cycle. 47 Adult-onset hypothyroidism induced a reduction in the number of proliferating cells and proliferative clusters in the SGZ as soon as 20 days after thyroidectomy, the first period studied, and also 1 month later. We observed the same reduction in newborn neuroblasts.…”

Section: Discussionmentioning

confidence: 99%

Modulation of adult hippocampal neurogenesis by thyroid hormones: implications in depressive-like behavior

et al. 2006

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Hormonal imbalances are involved in many of the age-related pathologies, as neurodegenerative and psychiatric diseases. Specifically, thyroid state alterations in the adult are related to psychological changes and mood disorders as depression. The dentate gyrus of the hippocampal formation undergoes neurogenesis in adult mammals including humans. Recent evidence suggests that depressive disorders and their treatment are tightly related to the number of newly born neurons in the dentate gyrus. We have studied the effect of thyroid hormones (TH) on hippocampal neurogenesis in adult rats in vivo. A short period of adultonset hypothyroidism impaired normal neurogenesis in the subgranular zone of the dentate gyrus with a 30% reduction in the number of proliferating cells. Hypothyroidism also reduced the number of newborn neuroblasts and immature neurons (doublecortin (DCX) immunopositive cells) which had a severely hypoplastic dendritic arborization. To correlate these changes with hippocampal function, we subjected the rats to the forced swimming and novel object recognition tests. Hypothyroid rats showed normal memory in object recognition, but displayed abnormal behavior in the forced swimming test, indicating a depressive-like disorder. Chronic treatment of hypothyroid rats with TH not only normalized the abnormal behavior but also restored the number of proliferative and DCX-positive cells, and induced growth of their dendritic trees. Therefore, hypothyroidism induced a reversible depressive-like disorder, which correlated to changes in neurogenesis. Our results indicate that TH are essential for adult hippocampal neurogenesis and suggest that mood disorders related to adult-onset hypothyroidism in humans could be due, in part, to impaired neurogenesis.

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“…Immunofluoresence for BrdU was detected with either AlexaFluor ® 488 or 594. Proliferation was also identified with a mouse monoclonal antiserum for (1:100; NCL-L-Ki67-MM1, Nova Castra, Newcastle upon Tyne, UK) (Kee et al, 2002), a mouse-on-mouse peroxidase kit (MOM ™ , Vector Labs) as visualized by DAB. Apoptotic cells were identified by terminal deoxynucleotidyl transferase-mediated dUTPbiotin in situ end-labeling (TUNEL) using a commercially available kit (Intergen, Purchase, NY, USA).…”

Section: Morphological Examinationmentioning

confidence: 99%

Trimethyltin-induced neurogenesis in the murine hippocampus

Harry¹,

McPherson²,

Wine³

et al. 2003

neurotox res

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Neurogenesis continues to occur in the mature rodent brain with one of the most prominent sources for new neurons being the subgranular layer (SGL) of the dentate gyrus (DG) in the hippocampus. A number of factors can stimulate this process including synaptic activity and injury. To determine if this process would occur upon a direct injury to the dentate region, we exposed young, 21 day old male CD-1 mice to the hippocampal toxicant, trimethyltin (TMT). An acute i.p. injection of TMT (2 mg/kg) produced extensive damage and loss of dentate granule neurons within 72 h. This active period of degeneration was accompanied by an increase in the generation of progenitor cells within the SGL as identified by BrdU uptake and Ki-67 immunostaining. As additional markers for neurogenesis, both nestin and doublecortin showed increased staining patterns within the blades of the dentate. In these young weanling mice, the level of proliferation was sufficient to significantly repopulate the dentate region by 4 weeks post-TMT, suggesting a high level of regenerative potential. Our data indicate a significant level of neurogenesis occurring during the active process of degeneration and in an environment of microglia activation. The TMT-induced injury offers a model system for further examination of the process of neurogenesis, neural adaptation, and the influence of inflammatory factors and glia interactions.

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The utility of Ki-67 and BrdU as proliferative markers of adult neurogenesis

Cited by 682 publications

References 30 publications

Bcl-XL modulates the differentiation of immortalized human neural stem cells

Bcl-XL modulates the differentiation of immortalized human neural stem cells

Modulation of adult hippocampal neurogenesis by thyroid hormones: implications in depressive-like behavior

Trimethyltin-induced neurogenesis in the murine hippocampus

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