the-subventricular-zone-en-face-wholemount-staining-and-ependymal-flow

The Subventricular Zone En-face: Wholemount Staining and Ependymal Flow

The walls of the lateral ventricles contain the largest germinal region in the adult mammalian brain. The subventricular zone (SVZ) in these walls is an ...

The lateral walls of the forebrain lateral ventricles are the richest source of stem cells in the adult mammalian brain. These stem cells give rise to new olfactory neurons that are renewed throughout life. The neurons originate in the subventricular zone (SVZ), migrate within the rostral extension (RE) of the SVZ along the rostral migratory stream (RMS) within tube-like structures formed of glial cells, to eventually reach the olfactory bulb (OB). We demonstrate that, contrary to the current view, multipotential (neuronal-astroglial-oligodendroglial) precursors with stem cell features can be isolated not only from the SVZ but also from the entire RE, including the distal portion within the OB. Specifically, these stem cells do not derive from the migratory neuroblasts coming from the SVZ. Interestingly, stem cells isolated from the proximal RE generate significantly more oligodendrocytes, and those from the distal RE proliferate significantly more slowly than stem cells derived from the SVZ and other RE regions. These findings demonstrate that stem cells are not confined to the forebrain periventricular region and indicate that stem cells endowed with different functional characteristics occur at different levels of the SVZ-RE pathway.

Multipotent neural stem cells reside into the rostral extension and olfactory bulb of adult rodents.

Multipotenten neuralen Stammzellen in der rostralen Erweiterung und Riechkolben von erwachsenen Nagern befinden.

The subventricular zone (SVZ) is the largest germinal layer in the adult mammalian brain and comprises stem cells, transit-amplifying progenitors, and committed neuroblasts. Although the SVZ contains the highest concentration of dividing cells in the adult brain, the intracellular mechanisms controlling their proliferation have not been elucidated. We show here that loss of the cyclin-dependent kinase inhibitor p27Kip1 has very specific effects on a population of CNS progenitors responsible for adult neurogenesis. Using bromodeoxyuridine and [(3)H]thymidine incorporation to label cells in S phase and cell-specific markers and electron microscopy to identify distinct cell types, we compared the SVZ structure and proliferation characteristics of wild-type and p27Kip1-null mice. Loss of p27Kip1 had no effect on the number of stem cells but selectively increased the number of the transit-amplifying progenitors concomitantly with a reduction in the number of neuroblasts. We conclude that cell-cycle regulation of SVZ adult progenitors is remarkably cell-type specific, with p27Kip1 being a key regulator of the cell division of the transit-amplifying progenitors.

Lack of the cell-cycle inhibitor p27Kip1 results in selective increase of transit-amplifying cells for adult neurogenesis.

Das Fehlen des Zellzyklus-Inhibitor p27Kip1 f&#xFC;hrt zur selektiven Erh&#xF6;hung der Transit-verst&#xE4;rkenden Zellen f&#xFC;r die adulte Neurogenese.

Genetics of childhood disorders: XXXVIII. Stem cell research, part 2: reconstructing the brain.

Genetik der Kindheit St&#xF6;rungen: XXXVIII. Stammzellforschung, Teil 2: Rekonstruktion des Gehirns.

Neurogenesis continues into adult life in restricted germinal layers. The identification of the neural stem cells that give rise to these new neurons has important clinical implications and provides fundamental information to understand the origins of the new neurons. Work in adult birds and rodents yielded a surprising result: the neural stem cells appear to have characteristics of glia. In adult birds, the primary neuronal precursors are radial glia. In adult mammals, the primary neuronal precursors have properties of astrocytes. Radial glial cells have previously been shown to transform into astrocytes; both cell types are classically considered part of a committed astroglial lineage. Instead, we propose that neural stem cells are contained within this astroglial lineage. These findings in adult vertebrate brain, together with recent work in the developing mammalian cerebral cortex, force us to reexamine traditional concepts about the origin of neurons and glia in the central nervous system. In particular, neural stem cells possess a surprisingly elaborate structure, suggesting that in addition to their progenitor role, they have important structural and metabolic support functions. The very same cells that give birth to new neurons also seem to nurture their maturation and support their function.

Identification of neural stem cells in the adult vertebrate brain.

Identifizierung von neuronalen Stammzellen im erwachsenen Gehirn von Wirbeltieren.

Neural stem cells in the subventricular zone (SVZ) continue to generate new neurons in the adult brain. SVZ cells exposed to EGF in culture grow to form neurospheres that are multipotent and self-renewing. We show here that the majority of these EGF-responsive cells are not derived from relatively quiescent stem cells in vivo, but from the highly mitotic, Dlx2(+), transit-amplifying C cells. When exposed to EGF, C cells downregulate Dlx2, arrest neuronal production, and become highly proliferative and invasive. Killing Dlx2(+) cells dramatically reduces the in vivo response to EGF and neurosphere formation in vitro. Furthermore, purified C cells are 53-fold enriched for neurosphere generation. We conclude that transit-amplifying cells retain stem cell competence under the influence of growth factors.

EGF converts transit-amplifying neurogenic precursors in the adult brain into multipotent stem cells.

EGF konvertiert Transit-verst&#xE4;rkenden neurogenen Vorl&#xE4;ufer im erwachsenen Gehirn in multipotente Stammzellen.

The adult mammalian brain harbors multipotent stem cells, which reside and participate in specialized niches that support self-renewal and differentiation. The first cellular and molecular elements of the stem cell niche in the adult brain have been identified and include cell-cell interactions and somatic cell signaling, the vasculature, the extracellular matrix and basal lamina. Furthermore, regulation at the epigenetic level via chromatin modification and remodeling is an integral aspect of stem cell biology. Understanding the in vivo stem cell niche will provide a framework for the elucidation of stem cell function in the adult brain.

A niche for adult neural stem cells.

Eine Nische f&#xFC;r adulten neuralen Stammzellen.

Glia are the most numerous cells in the brain, and their many diverse functions highlight their essential role in the nervous system. Recent studies have revealed an unexpected new role for glia in a wide variety of species, that of stem cells/progenitors in the adult and embryonic brain. Differentiation along the glial lineage may be a default state of development reflected in the progression of stem cells along the neuroepithelial-->radial glia-->astrocyte lineage.

The glial identity of neural stem cells.

Die Glia-Identit&#xE4;t von neuralen Stammzellen.

Adult neurogenesis occurs in most species and is regulated by a wide variety of environmental and pharmacological challenges. The functional integration of neurons generated in the adult was first demonstrated in songbirds more than two decades ago. In the adult mammalian brain, neurons are continuously generated in two structures, the olfactory bulb and the hippocampus. Current evidence suggests that adult-born immature neurons have distinct electrophysiological properties from old neurons, and proposed roles in a variety of functions including olfaction, learning and mood regulation.

Young and excitable: the function of new neurons in the adult mammalian brain.

Junge und erregbar: Die Funktion von neuen Nervenzellen im erwachsenen Gehirn von S&#xE4;ugetieren.

The complexity and cellular diversity of the adult brain arises from the proliferation and differentiation of a small number of stem cells. The intrinsic state of stem cells depends on their spatial and temporal history and affects their responsiveness to extrinsic signals from the microenvironment. Stem cell self-renewal and differentiation along neuronal and glial lineages are defined by the dynamic interplay between transcription, epigenetic control, and posttranscriptional regulators, including microRNAs, whose key role in stem cell biology is just emerging.

Stem cells: from epigenetics to microRNAs.

Stammzellen: aus der Epigenetik zu microRNAs.

Neurogenesis persists in two germinal regions in the adult mammalian brain, the subventricular zone of the lateral ventricles and the subgranular zone in the hippocampal formation. Within these two neurogenic niches, specialized astrocytes are neural stem cells, capable of self-renewing and generating neurons and glia. Cues within the niche, from cell-cell interactions to diffusible factors, are spatially and temporally coordinated to regulate proliferation and neurogenesis, ultimately affecting stem cell fate choices. Here, we review the components of adult neural stem cell niches and how they act to regulate neurogenesis in these regions.

Brain micro-ecologies: neural stem cell niches in the adult mammalian brain.

Neurogenese beibehalten in zwei germinal Regionen in das Erwachsene Gehirn von S&auml;ugetieren, die subventrikul&auml;re Zone von der Seitenventrikel und subgranular Zone in der hippocampal Formation. Innerhalb dieser zwei neurogenen Nischen sind spezialisierte Astrozyten neurale Stammzellen, selbstverl&auml;ngerndes und Generieren von Neuronen und Glia. Hinweise in der Nische, aus Zell-Zell-Interaktionen diffusible Faktoren, werden r&auml;umlich und zeitlich koordiniert, Proliferation und Neurogenese, letztlich beeintr&auml;chtigen Stammzellen Schicksal Entscheidungen zu regulieren. Hier &uuml;berpr&uuml;fen wir die Komponenten der adulten neuralen Stammzellen Nischen und wie sie handeln, um die Neurogenese in diesen Regionen zu regulieren.

Mikro-&Ouml;kologien des Gehirns: Neuronale Stammzellen Nischen im erwachsenen Gehirn von S&auml;ugetieren.

Stem cells (SC) exhibit a unique capacity for self-renewal in an undifferentiated state. It is unclear whether the self-renewal of pluripotent embryonic SC (ESC) and of tissue-specific adult SC such as hematopoietic SC (HSC) is controlled by common mechanisms. The deletion of transcription factor Zfx impaired the self-renewal but not the differentiation capacity of murine ESC; conversely, Zfx overexpression facilitated ESC self-renewal by opposing differentiation. Furthermore, Zfx deletion abolished the maintenance of adult HSC but did not affect erythromyeloid progenitors or fetal HSC. Zfx-deficient ESC and HSC showed increased apoptosis and SC-specific upregulation of stress-inducible genes. Zfx directly activated common target genes in ESC and HSC, as well as ESC-specific target genes including ESC self-renewal regulators Tbx3 and Tcl1. These studies identify Zfx as a shared transcriptional regulator of ESC and HSC, suggesting a common genetic basis of self-renewal in embryonic and adult SC.

Zfx controls the self-renewal of embryonic and hematopoietic stem cells.

Stammzellen (SC) weisen eine einzigartige Kapazit&auml;t f&uuml;r Selbsterneuerung in einem undifferenzierten Zustand. Es ist unklar, ob der Selbsterneuerung von pluripotenten embryonalen SC (ESC) und der Gewebe-spezifischen Erwachsene SC wie h&auml;matopoetische SC (HSC) durch gemeinsame Mechanismen gesteuert. Die Streichung des Transkriptionsfaktors Zfx beeintr&auml;chtigt die Selbsterneuerung aber nicht die Differenzierung-Kapazit&auml;t der murine ESC; umgekehrt erleichtert Zfx &Uuml;berexpression ESC Selbsterneuerung abzielende Differenzierung. Au&szlig;erdem Zfx L&ouml;schung die Aufrechterhaltung der Erwachsenen HSC abgeschafft aber hatte keine Auswirkungen auf Erythromyeloid Vorfahre oder fetalen HSC. ZFX-defizienten ESC und HSC zeigte erh&ouml;hte Apoptose und SC-spezifische Hochregulation der Spannung-inducible Gene. ZFX aktiviert direkt gemeinsame Zielgene in ESC und HSC sowie ESC-spezifischen Zielgene einschlie&szlig;lich ESC Selbsterneuerung Regulierungsbeh&ouml;rden Tbx3 und Tcl1. Diese Studien identifizieren Zfx als einen freigegebenen transcriptional Regulator der ESC und HSC, was auf eine gemeinsame genetische Grundlage der Selbsterneuerung in embryonalen und adulten SC.

ZFX steuert der Selbsterneuerung von embryonalen und h&auml;matopoetischen Stammzellen.

Stem cells reside in specialized niches that regulate their self-renewal and differentiation. The vasculature is emerging as an important component of stem cell niches. Here, we show that the adult subventricular zone (SVZ) neural stem cell niche contains an extensive planar vascular plexus that has specialized properties. Dividing stem cells and their transit-amplifying progeny are tightly apposed to SVZ blood vessels both during homeostasis and regeneration. They frequently contact the vasculature at sites that lack astrocyte endfeet and pericyte coverage, a modification of the blood-brain barrier unique to the SVZ. Moreover, regeneration often occurs at these sites. Finally, we find that circulating small molecules in the blood enter the SVZ. Thus, the vasculature is a key component of the adult SVZ neural stem cell niche, with SVZ stem cells and transit-amplifying cells uniquely poised to receive spatial cues and regulatory signals from diverse elements of the vascular system.

A specialized vascular niche for adult neural stem cells.

Stammzellen befinden sich in speziellen Nischen, die ihre Selbsterneuerung und Differenzierung regulieren. Das Gef&auml;&szlig;system ist als wichtiger Bestandteil der Stammzelle Nischen entstehen. Hier zeigen wir, dass die erwachsenen subventrikul&auml;re Zone (SVZ) neuronalen Stammzelle Nische eine umfangreiche planar vaskul&auml;ren Plexus enth&auml;lt, die Eigenschaften spezialisiert hat. Teilung von Stammzellen und ihre Nachkommen Transit-Verst&auml;rkung sind eng apposed SVZ Blutgef&auml;&szlig;e, sowohl w&auml;hrend der Hom&ouml;ostase und Regeneration. Sie wenden h&auml;ufig das Gef&auml;&szlig;system an Standorten, die Gliazelle-Endfeet und Pericyt-Abdeckung, eine &Auml;nderung der die Blut - Hirn Schranke eindeutig die SVZ abgeht. Dar&uuml;ber hinaus tritt die Regenerierung oft an diesen St&auml;tten. Schlie&szlig;lich finden wir, dass die zirkulierenden kleine Molek&uuml;le im Blut die SVZ eingeben. Somit ist das Gef&auml;&szlig;system eine Schl&uuml;sselkomponente der Erwachsenen SVZ neuronalen Stammzelle Nische, mit SVZ Stammzellen und Transit-Verst&auml;rkung Zellen eindeutig balanciert, um r&auml;umliche Cues und regulatorische Signale von verschiedenen Elemente des Gef&auml;&szlig;systems erhalten.

Eine spezielle vaskul&auml;re Nische f&uuml;r adulte neurale Stammzellen.

The subventricular zone (SVZ) is the largest neurogenic niche in the adult mammalian brain. We found that the brain-enriched microRNA miR-124 is an important regulator of the temporal progression of adult neurogenesis in mice. Knockdown of endogenous miR-124 maintained purified SVZ stem cells as dividing precursors, whereas ectopic expression led to precocious and increased neuron formation. Furthermore, blocking miR-124 function during regeneration led to hyperplasias, followed by a delayed burst of neurogenesis. We identified the SRY-box transcription factor Sox9 as being a physiological target of miR-124 at the transition from the transit amplifying cell to the neuroblast stage. Sox9 overexpression abolished neuronal differentiation, whereas Sox9 knockdown led to increased neuron formation. Thus miR-124-mediated repression of Sox9 is important for progression along the SVZ stem cell lineage to neurons.

miR-124 regulates adult neurogenesis in the subventricular zone stem cell niche.

Die subventrikul&auml;re Zone (SVZ) ist die gr&ouml;&szlig;te neurogene Nische im erwachsenen Gehirn von S&auml;ugetieren. Wir fanden, dass das Gehirn angereicherten MicroRNA MiR-124 ein wichtiger Regulator der zeitlichen Progression der adulten Neurogenese bei M&auml;usen ist. Niederschlag des endogenen MiR-124 beibehalten gereinigten SVZ-Stammzellen als Vorl&auml;ufer, Division, w&auml;hrend ektopische Expression zu fr&uuml;hreif und erh&ouml;hte Neuron Bildung gef&uuml;hrt. Dar&uuml;ber hinaus, blockieren MiR-124-Funktion w&auml;hrend der Regenerierung f&uuml;hrte zu Hyperplasias, gefolgt von einer verz&ouml;gerten Platzen der Neurogenese. Wir identifizieren den SRY-Box-Transkriptionsfaktor Sox9 als eine physiologische Ziel von MiR-124 am &Uuml;bergang von der Transit Verst&auml;rkung der Zelle auf die B&uuml;hne Blast (Biologie). Sox9-&Uuml;berexpression abgeschafft neuronale Differenzierung, w&auml;hrend Sox9 Niederschlag zu erh&ouml;hten Neuron Bildung gef&uuml;hrt. MiR-124-vermittelter Unterdr&uuml;ckung von Sox9 ist somit wichtig f&uuml;r Weiterentwicklung entlang der SVZ Stammzell-&Uuml;bertragungslinie zu Neuronen.

MiR-124 regelt adulten Neurogenese in der subventrikul&auml;re Zone Stammzelle Nische.

The ability to prospectively isolate adult neural stem cells and their progeny is crucial to study their biology and therapeutic potential. Stem cells in adult mammalian neurogenic niches are a subset of astrocytes. A major limitation in the field has been the inability to distinguish stem cell astrocytes from niche astrocytes. Here, we show that epidermal growth factor receptor (EGFR)-positive subventricular-zone (SVZ) astrocytes are activated stem cells that are eliminated by antimitotic treatment. We developed a simple strategy to simultaneously purify cells at different stages of the adult SVZ stem cell lineage by using FACS. This method combines the use of fluorescent EGF ligand, CD24, and GFP expression in GFAP::GFP transgenic mice and allows the simultaneous purification of activated stem cell astrocytes (GFP(+)EGFR(+)CD24(-)), niche astrocytes (GFP(+)EGFR(-)CD24(-)), transit amplifying cells (GFP(-)EGFR(+)CD24(-)), and neuroblasts (GFP(-)EGFR(-)CD24(low)). One in three EGFR(+) astrocytes gives rise to neurospheres in vitro, a 20-fold enrichment over unsorted cells. Importantly, these cells constitute the neurosphere-forming population among SVZ astrocytes. This approach will be of great utility for future functional and molecular studies of the SVZ stem cell lineage.

Simultaneous prospective purification of adult subventricular zone neural stem cells and their progeny.

Die F&auml;higkeit, prospektiv adulten neuralen Stammzellen und ihre Nachkommen zu isolieren ist entscheidend f&uuml;r ihre Biologie und therapeutisches Potential zu studieren. Stammzellen in Erwachsenen S&auml;ugetier-neurogene Nischen sind eine Teilmenge der Astrozyten. Eine wichtige Einschr&auml;nkung im Bereich wurde die Unf&auml;higkeit, Nische Astrozyten Stammzelle Astrozyten unterscheiden. Hier zeigen wir die epidermalen Wachstumsfaktor-Rezeptor (EGFR)-positive subventrikul&auml;re Zone (SVZ) Astrozyten sind aktivierte Stammzellen, die durch antimitotic Behandlung beseitigt werden. Wir entwickelten eine einfache Strategie um Zellen gleichzeitig in verschiedenen Phasen der Erwachsenen SVZ Stammzelle Abstammung zu reinigen, mithilfe von FACS. Diese Methode kombiniert die Verwendung von fluoreszierenden EGF-Ligand, CD24 und GFP Ausdruck in GFAP::GFP transgene M&auml;use und erm&ouml;glicht die gleichzeitige Reinigung von aktivierten Stammzelle Astrozyten (GFP(+)EGFR(+)CD24(-)), Nische Astrozyten (GFP(+)EGFR(-)CD24(-)), Transit, die Verst&auml;rkung der Zellen (GFP(-)EGFR(+)CD24(-)) und Neuroblasten (GFP(-)EGFR(-)CD24(low)). Einer von drei EGFR(+) Astrozyten gibt Grund zu Neurospheres in vitro eine zwanzigfache Bereicherung &uuml;ber unsortierten Zellen. Wichtig ist, bilden diese Zellen Neurosphere-bildenden Bev&ouml;lkerung unter SVZ Astrozyten. Dieser Ansatz wird von gro&szlig;em Nutzen f&uuml;r zuk&uuml;nftige funktionelle und Molekulare Studien der SVZ Stammzelle Abstammung sein.

Gleichzeitige prospektive Reinigung von erwachsenen subventrikul&auml;re Zone neurale Stammzellen und ihre Nachkommen.

The inference of transcriptional networks that regulate transitions into physiological or pathological cellular states remains a central challenge in systems biology. A mesenchymal phenotype is the hallmark of tumour aggressiveness in human malignant glioma, but the regulatory programs responsible for implementing the associated molecular signature are largely unknown. Here we show that reverse-engineering and an unbiased interrogation of a glioma-specific regulatory network reveal the transcriptional module that activates expression of mesenchymal genes in malignant glioma. Two transcription factors (C/EBPbeta and STAT3) emerge as synergistic initiators and master regulators of mesenchymal transformation. Ectopic co-expression of C/EBPbeta and STAT3 reprograms neural stem cells along the aberrant mesenchymal lineage, whereas elimination of the two factors in glioma cells leads to collapse of the mesenchymal signature and reduces tumour aggressiveness. In human glioma, expression of C/EBPbeta and STAT3 correlates with mesenchymal differentiation and predicts poor clinical outcome. These results show that the activation of a small regulatory module is necessary and sufficient to initiate and maintain an aberrant phenotypic state in cancer cells.

The transcriptional network for mesenchymal transformation of brain tumours.

Die Folgerung transkriptionelle Netze, die &Uuml;berg&auml;nge in physiologischen oder pathologischen zellul&auml;ren Staaten Regeln bleibt eine zentrale Herausforderung in Systembiologie. Ein mesenchymaler Ph&auml;notyp ist das Markenzeichen der Tumor Aggressivit&auml;t in menschlichen malignes Gliom, aber die rechtlichen Programme f&uuml;r die Durchf&uuml;hrung der zugeordneten molekularen Signatur sind weitgehend unbekannt. Hier zeigen wir, dass Reverse Engineering und eine unvoreingenommene Verh&ouml;r eines Gliom-spezifischen regulatorischen Netzes offenbaren das transkriptionelle Modul, das Ausdruck der Mesenchymale Gene in malignes Gliom aktiviert. Zwei Transkriptionsfaktoren (C/EBPbeta und STAT3) entstehen als synergistische Initiatoren und master Regler der mesenchymalen Transformation. Ektopische Koexpression von C/EBPbeta und STAT3 reprograms neuralen Stammzellen entlang der abweichenden mesenchymal-Linie, w&auml;hrend die Beseitigung der beiden Faktoren in Gliomzellen f&uuml;hrt zum Zusammenbruch der Mesenchymale Signatur und reduziert den Tumor Aggressivit&auml;t. In menschlichen Gliom Ausdruck von C/EBPbeta und STAT3 korreliert mit Mesenchymale Differenzierung und ung&uuml;nstigen klinischen Verlauf prognostiziert. Diese Ergebnisse zeigen, dass die Aktivierung eines kleinen regulatorischen Moduls ist notwendig und ausreichend zu initiieren und pflegen einen aberrante ph&auml;notypischen Zustand in Krebszellen.

Das transkriptionelle Netzwerk f&uuml;r Mesenchymale Transformation von Hirntumoren.

We investigated the expression and role of the dopamine receptor 3 (D3R) in postnatal mouse subventricular zone (SVZ). In situ hybridization detected selective D3R mRNA expression in the SVZ. Fluorescence activated cell sorting (FACS) of adult SVZ subtypes using hGFAP-GFP and Dcx-GFP mice showed that transit amplifying progenitor cells and niche astrocytes expressed D3R whereas stem cell-like astrocytes and neuroblasts did not. To determine D3R's role in SVZ neurogenesis, we administered U-99194A, a D3R preferential antagonist, and bromodeoxyuridine in postnatal mice. In vivo D3R antagonism decreased the numbers of newborn neurons reaching the core and the periglomerular layer of the olfactory bulb. Moreover, it decreased progenitor cell proliferation but did not change the number of label-retaining (stem) cells, commensurate with its expression on transit amplifying progenitor cells but not SVZ stem cell-like astrocytes. Collectively, this study suggests that dopaminergic stimulation of D3R drives proliferation via rapidly amplifying progenitor cells to promote murine SVZ neurogenesis.

Dopamine stimulation of postnatal murine subventricular zone neurogenesis via the D3 receptor.

Wir untersuchten den Ausdruck und die Rolle des Dopamin-Rezeptors 3 (D3R) bei postnatalen Maus subventrikul&auml;re Zone (SVZ). In situ Hybridisierung erkannt selektive D3R mRNA Ausdruck in der SVZ. Fluoreszenz aktivierte Zellsortierung (FACS) von Erwachsenen SVZ-Subtypen, die mit hGFAP-GFP und Dcx-GFP-M&auml;usen zeigten, dass Transit Kursst&uuml;rzen Vorl&auml;uferzellen und Nische Astrozyten D3R ausgedr&uuml;ckt, w&auml;hrend die Stammzell-wie Astrozyten und Neuroblasten nicht tat. Um D3Rs Rolle in SVZ Neurogenese bestimmen, verwaltet wir U-99194A, D3R pr&auml;ferenzielle Antagonist und Bromdesoxyuridin bei postnatalen M&auml;usen. In-vivo D3R Antagonismus sank die Zahl der neugeborenen Neuronen, die den Kern und die Periglomerular Schicht der Riechkolben erreichen. Dar&uuml;ber hinaus verringerte Vorl&auml;uferzellen Zellproliferation es aber &auml;nderte nicht die Anzahl der Etikett-behalten () Stammzellen, angemessen mit seinen Ausdruck auf Transit amplifying Vorl&auml;uferzellen aber nicht SVZ Stammzell-wie Astrozyten. Kollektiv, schl&auml;gt dieser Studie dieser dopaminergen Stimulation von D3R Laufwerke Verbreitung &uuml;ber rasch Verst&auml;rkung Vorl&auml;uferzellen murinen SVZ Neurogenese zu f&ouml;rdern.

Dopamin-Stimulation der postnatalen murinen subventrikul&auml;re Zone Neurogenese &uuml;ber den D3-Rezeptor.

Sphere-forming assays have been widely used to retrospectively identify stem cells based on their reported capacity to evaluate self-renewal and differentiation at the single-cell level inÂ vitro. The discovery of markers that allow the prospective isolation of stem cells and their progeny from their inÂ vivo niche allows the functional properties of purified populations to be defined. We provide a historical perspective of the evolution of the neurosphere assay and highlight limitations in the use of sphere-forming assays in the context of neurospheres. We discuss theoretical and technical considerations of experimental design and interpretation that surround the use of this assay with any tissue.

Eyes wide open: a critical review of sphere-formation as an assay for stem cells.

Kugel-bildenden Proben haben verbreitet, um Stammzellen, die auf der Grundlage ihrer gemeldeten F&auml;higkeit zur Selbsterneuerung und Differenzierung auf in-vitro-Einzeller-Ebene bewerten r&uuml;ckwirkend zu identifizieren. Die Entdeckung von Markern, mit denen die potentielle Isolation der Stammzellen und ihre Nachkommen von ihrer in-vivo-Nische kann die funktionalen Eigenschaften der gereinigten Populationen definiert werden. Wir bieten einen historischen Perspektive der Entwicklung der Neurosphere-Assay und betonen Einschr&auml;nkungen bei der Verwendung von Kugel-bildenden Proben im Rahmen des Neurospheres. Wir diskutieren die theoretische und technische &Uuml;berlegungen der experimentellen Gestaltung und Auslegung, die die Verwendung dieses Testsystems mit jedem m&ouml;glichem Gewebe umgeben.

Article	Year
Multipotent neural stem cells reside into the rostral extension and olfactory bulb of adult rodents. The Journal of neuroscience : the official journal of the Society for Neuroscience\| PubMed ID: 11784788	2002
Lack of the cell-cycle inhibitor p27Kip1 results in selective increase of transit-amplifying cells for adult neurogenesis. The Journal of neuroscience : the official journal of the Society for Neuroscience\| PubMed ID: 11896165	2002
Genetics of childhood disorders: XXXVIII. Stem cell research, part 2: reconstructing the brain. Journal of the American Academy of Child and Adolescent Psychiatry\| PubMed ID: 12014796	2002
Identification of neural stem cells in the adult vertebrate brain. Brain research bulletin\| PubMed ID: 12031271	2002
EGF converts transit-amplifying neurogenic precursors in the adult brain into multipotent stem cells. Neuron\| PubMed ID: 12495619	2002
A niche for adult neural stem cells. Current opinion in genetics & development\| PubMed ID: 14550422	2003
The glial identity of neural stem cells. Nature neuroscience\| PubMed ID: 14583753	2003
Young and excitable: the function of new neurons in the adult mammalian brain. Current opinion in neurobiology\| PubMed ID: 15721754	2005
Stem cells: from epigenetics to microRNAs. Neuron\| PubMed ID: 15882632	2005
Brain micro-ecologies: neural stem cell niches in the adult mammalian brain. Philosophical transactions of the Royal Society of London. Series B, Biological sciences\| PubMed ID: 17322003	2008
Zfx controls the self-renewal of embryonic and hematopoietic stem cells. Cell\| PubMed ID: 17448993	2007
A specialized vascular niche for adult neural stem cells. Cell stem cell\| PubMed ID: 18786415	2008
miR-124 regulates adult neurogenesis in the subventricular zone stem cell niche. Nature neuroscience\| PubMed ID: 19287386	2009
Simultaneous prospective purification of adult subventricular zone neural stem cells and their progeny. Proceedings of the National Academy of Sciences of the United States of America\| PubMed ID: 19332781	2009
The transcriptional network for mesenchymal transformation of brain tumours. Nature\| PubMed ID: 20032975	2010
Dopamine stimulation of postnatal murine subventricular zone neurogenesis via the D3 receptor. Journal of neurochemistry\| PubMed ID: 20477937	2010
Eyes wide open: a critical review of sphere-formation as an assay for stem cells. Cell stem cell\| PubMed ID: 21549325	2011

Fiona Doetsch

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