A cytochemical and autoradiographic study of nuclear DNA in mouse Purkinje cells

Mareš, V; Lodin, Z; Sacha, Jonah B.

doi:10.1016/0006-8993(73)90214-x

Cited by 51 publications

(19 citation statements)

References 19 publications

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“…Moreover, applying fluorescence in situ hybridization (FISH) technique to paraffin sections of the adult human CNS, Yang et al (2001) could provide direct evidence for aneuploidy in AD, but not in normal brain. More recently, however, two groups (Rehen et al, 2005;Yurov et al, 2005) applying FISH to sorted isolated cellular nuclei reported a considerable degree of constitutional aneuploidy in the normal adult human brain, including triploidy and tetraploidy, corroborating cytophotometric observations made several decades ago (Brodskij and Kusc, 1962;Lapham and Johnstone, 1963;Mares et al, 1973). This kind of constitutional trisomy and tetrasomy might arise at least in part through chromosome mis-segregation during mitosis in neuronal progenitor cells and can persist into adulthood (A. H. .…”

Section: Introductionmentioning

confidence: 55%

Aneuploidy and DNA Replication in the Normal Human Brain and Alzheimer's Disease

Mosch

Morawski²,

Mittag³

et al. 2007

Journal of Neuroscience

240

248

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Reactivation of the cell cycle, including DNA replication, might play a major role in Alzheimer's disease (AD). A more than diploid DNA content in differentiated neurons might alternatively result from chromosome mis-segregation during mitosis in neuronal progenitor cells. It was our objective to distinguish between these two mechanisms for aneuploidy and to provide evidence for a functional cell cycle in AD. Using slide-based cytometry, chromogenic in situ hybridization, and PCR amplification of alu-repeats, we quantified the DNA amount of identified cortical neurons in normal human brain and AD and analyzed the link between a tetraploid DNA content and expression of the early mitotic marker cyclin B1. In the normal brain, the number of neurons with a more than diploid content amounts to ϳ10%. Less than 1% of neurons contains a tetraploid DNA content. These neurons do not express cyclin B1, most likely representing constitutional tetraploidy. This population of cyclin B1-negative tetraploid neurons, at a reduced number, is also present in AD. In addition, a population of cyclin B1-positive tetraploid neurons of ϳ2% of all neurons was observed in AD. Our results indicate that at least two different mechanisms need to be distinguished giving rise to a tetraploid DNA content in the adult brain. Constitutional aneuploidy in differentiated neurons might be more frequent than previously thought. It is, however, not elevated in AD. In addition, in AD some neurons have re-entered the cell cycle and entirely passed through a functional interphase with a complete DNA replication.

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Section: Introductionmentioning

confidence: 55%

Aneuploidy and DNA Replication in the Normal Human Brain and Alzheimer's Disease

Mosch

Morawski²,

Mittag³

et al. 2007

Journal of Neuroscience

240

248

View full text Add to dashboard Cite

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“…Reports of aneuploid Purkinje neurons being found within cerebella from aged rats and mice have been in the literature for quite some time (Del Monte, 2006; Mann et al, 1978; Mares et al, 1971; Lapham, 1968) and it is therefore reasonable to propose that they are the predominant fusogenic-capable CNS cell type able to form heterokaryons with BMDCs/HSCs. Furthermore, these binucleated heterokaryons were shown to be stable and viable in a long term study (Magrassi et al, 2007) and suggest selective cellular fusion offers considerable therapeutic potential in terms of showing promise for rescuing specific cells, especially in the inherited ataxias.…”

Section: Discussionmentioning

confidence: 99%

Cellular fusion for gene delivery to SCA1 affected Purkinje neurons

Chen

Cruz

Lanuto

et al. 2011

Molecular and Cellular Neuroscience

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Cerebellar Purkinje neurons (PNs) possess a well characterized propensity to fuse with bone marrow-derived cells (BMDCs), producing heterokaryons with Purkinje cell identities. This offers the potential to rescue/repair at risk or degenerating PNs in the inherited ataxias, including Spinocerebellar Ataxia 1 (SCA1), by introducing therapeutic factors through BMDCs to potentially halt or reverse disease progression. In this study, we combined gene therapy and a stem cell-based treatment to attempt repair of at-risk PNs through cell-cell fusion in a Sca1154Q/2Q knock-in mouse model. BMDCs enriched for the hematopoietic stem cell (HSC) population were genetically modified using adeno-associated viral vector 7 (AAV7) to carry SCA1 modifier genes and transplanted into irradiated Sca1154Q/2Q mice. Binucleated Purkinje heterokaryons with sex-mismatched donor Y chromosomes were detected and successfully expressed the modifier genes in vivo. Potential effects of the new genome within Purkinje heterokaryons were evaluated using nuclear inclusions (NIs) as a biological marker to reflect possible modifications of the SCA1 disease process. An overall decrease in number of NIs and an increase in the number of surviving PNs were observed in treated Sca1154Q/2Q. Furthermore, Bergmann glia were found to have fusogenic potential with the donor population and reveal another potential route of therapeutic entry into at-risk cells of the SCA1 cerebellum. This study presents a first step towards a proof of principle that combines somatic cellular fusion events with a neuroprotective gene therapy approach for providing potential neuronal protection/repair in a variety of neurodegenerative disorders.

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“…Recent reports showed that under strong selective pressure in tissue culture, embryonic stem cells fused with either bone marrow cells or neural stem cells (16,17). Despite early studies that suggested they were tetraploid (18), Purkinje cells have now clearly been shown to be diploid (19)(20)(21). Consequently, analyses of sex chromosome composition could determine not only whether bone marrow contributed to Purkinje cells, but also provide insight into the underlying mechanism.…”

Section: Discussionmentioning

confidence: 99%

Contribution of transplanted bone marrow cells to Purkinje neurons in human adult brains

Weimann

Charlton²,

Brazelton³

et al. 2003

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

397

241

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We show here that cells within human adult bone marrow can contribute to cells in the adult human brain. Cerebellar tissues from female patients with hematologic malignancies, who had received chemotherapy, radiation, and a bone marrow transplant, were analyzed. Brain samples were obtained at autopsy from female patients who received male (sex-mismatched) or female (sexmatched, control) bone marrow transplants. Cerebella were evaluated in 10-m-thick, formaldehyde-fixed, paraffin-embedded sections that encompassed up to Ϸ50% of a human Purkinje nucleus. A total of 5,860 Purkinje cells from sex-mismatched females and 3,202 Purkinje cells from sex-matched females were screened for Y chromosomes by epifluorescence. Confocal laser scanning microscopy allowed definitive identification of the sex chromosomes within the morphologically distinct Purkinje cells. In the brains of females who received male bone marrow, four Purkinje neurons were found that contained an X and a Y chromosome and two other Purkinje neurons contained more than a diploid number of sex chromosomes. No Y chromosomes were detected in the brains of sex-matched controls. The total frequency of male bone marrow contribution to female Purkinje cells approximated 0.1%. This study demonstrates that although during human development Purkinje neurons are no longer generated after birth, cells within the bone marrow can contribute to these CNS neurons even in adulthood. The underlying mechanism may be caused either by generation de novo of Purkinje neurons from bone marrow-derived cells or by fusion of marrow-derived cells with existing recipient Purkinje neurons.stem cell ͉ plasticity ͉ cell fusion ͉ cell fate change I n humans, bone marrow has been reported to contribute to human epithelium and liver, but not to the brain (1, 2). Here we investigated whether bone marrow-derived cells could cross the blood-brain barrier and contribute to neurons in the CNS. Previous studies in mice have shown that bone marrow-derived cells can contribute to neuronal cell types in the CNS, including a class of highly specialized neurons in the brain, the Purkinje neurons (3-5).Purkinje neurons are generated only during early brain development. In humans, generation of Purkinje neurons starts at 16 weeks of gestation and is complete by the end of the 23rd week (6). Most of the maturation of the characteristic dendritic trees of human Purkinje neurons is finalized during the first year of life (7). By contrast to other neurons in the adult brain, there is no evidence for the generation of new Purkinje neurons after birth, even in cases of severe Purkinje cell loss caused by trauma or genetic disease (8, 9).The human brain contains Ϸ15 million Purkinje cells, which are among the largest neurons in the CNS (10). A typical Purkinje neuron has Ͼ50-fold the volume of neighboring neurons in the brain, and its complex dendritic extensions receive inputs from as many as one million granule cells. Purkinje cells play vital roles in maintaining balance and regulating movement. A loss o...

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A cytochemical and autoradiographic study of nuclear DNA in mouse Purkinje cells

Cited by 51 publications

References 19 publications

Aneuploidy and DNA Replication in the Normal Human Brain and Alzheimer's Disease

Aneuploidy and DNA Replication in the Normal Human Brain and Alzheimer's Disease

Cellular fusion for gene delivery to SCA1 affected Purkinje neurons

Contribution of transplanted bone marrow cells to Purkinje neurons in human adult brains

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