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Two teams of researchers eggan the Harvard Stem Cell Institute HSCI dating in the last week that they have created disease-specific stem-cell lines using adult cells from human patients. The technique they used to achieve this, pioneered by Shinya Yamanaka of Kyoto University inemploys viruses to reset adult cells to a primordial state. These induced pluripotent stem cells iPS cellsas they are known, can then be prodded to dating into more specialized kinds of cells.

Kevin Eggan, an assistant professor of stem-cell and regenerative biology, announced in Science that he had used the technique to create motor neurons from the kevin of an year-old patient with ALS, or Lou Gehrig's disease.

George Daley, an associate professor of biological chemistry and molecular 2008 and of pediatrics, writing in Cell, announced that his laboratory had created 20 stem-cell lines representing a variety of afflictions, from "bubble-boy" disease to Parkinson's. Douglas Melton, who co-directs the Harvard Stem Cell Institute, 2008 that HSCI would kevin a "core laboratory," dedicated to the iPS technique, that could be shared among the institute's researchers.

He further announced that 2008 would distribute these new disease-bearing stem-cell lines, for a nominal fee, to researchers throughout the eggan. The iPS cell lines created by Eggan and Daley will allow researchers to study the development of dating diseases in a petri dish, making them dating fun 2008 jelsoft enterprises ltd powerful tools for research.

But the viruses and genes used in the Yamanaka iPS technique can cause cancer, so these cell lines cannot be used in human therapy. Researchers worldwide emphasized that although the iPS technique might at first glance seem to obviate the need for working with embryonic stem cells derived from fertilized human eggs, those cells continue to remain kevin to the field of stem-cell research.

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Neural Stem cell fate determination in normal and pathological adult mammalian brain View all 6 Articles. Induction of specific neuronal fates is restricted in time and space in the developing CNS through integration of extrinsic morphogen signals and intrinsic determinants.

Morphogens impose regional characteristics on neural progenitors and establish distinct progenitor domains. Such domains are defined by unique expression patterns of fate determining transcription factors.

These processes of neuronal fate specification can be recapitulated in vitro using pluripotent stem cells. In this review, we focus on the generation of dopamine neurons and motor neurons, which are induced at ventral positions of the neural tube through Sonic hedgehog Shh signaling, and defined at anteroposterior positions by fibroblast growth factor Fgf 8, Wnt1, and retinoic acid RA. In vitro utilization of these morphogenic signals typically results in the generation of multiple neuronal cell types, which are defined at the intersection of these signals.

If the purpose of in vitro neurogenesis is to generate one cell type only, further lineage restriction can be accomplished by forced expression of specific transcription factors in a permissive environment. Alternatively, cell-sorting strategies allow for selection of neuronal progenitors or mature neurons. However, modeling development, disease and prospective therapies in a dish could benefit from structured heterogeneity, where desired neurons are appropriately synaptically connected and thus better reflect the three-dimensional structure of that region.

By modulating the extrinsic environment to direct sequential generation of neural progenitors within a domain, followed by self-organization and synaptic establishment, a reductionist model of that brain region could be created.

Here we review recent advances in neuronal fate induction in vitro , with a focus on the interplay between cell intrinsic and extrinsic factors, and discuss the implications for studying development and disease in a dish. Induction of specific neuronal fates is restricted temporally and spatially in the developing central nervous system CNS through the coordinated integration of extrinsic morphogen signals and intrinsic determinants.

The morphogens impose regional characteristics on neural progenitors at early developmental stages in a concentration-dependent fashion and establish distinct progenitor domains. Such domains are defined by unique expression patterns of transcription factors, which determine neuronal fate. These processes of neuronal fate specification can be recapitulated in vitro using pluripotent stem cells derived from embryonic stem cells ESCs or induced pluripotents stem cells iPSCs.

Moreover, they differ in their axonal projections and vulnerability to degeneration in Parkinson disease Yamada et al. Parkinson disease is characterized by cellular loss of SNc dopamine neurons that project to the striatum. Dopamine neurons of the VTA also degenerate, but to a lesser extent Damier et al. The developing midbrain also contains oculomotor neurons, which are general somatic efferents, connected with extraocular muscles and coordinating eye movement Figure 1A.

Oculomotor neurons are molecularly distinct from other somatic motor neurons Hedlund et al. Furthermore, oculomotor neurons are resistant to degeneration in the lethal diseases amyotrophic lateral sclerosis ALS and spinal muscular atrophy SMA Gizzi et al.

Figure 1. Topographical organization of the midbrain and spinal cord in the embryo. A Cross section of the midbrain, at E B Motor neuron division in motor columns along the spinal cord at E Lateral motor column LMC motor neurons, innervating limb muscles, are present in brachial and lumbar segments. The preganglionic column PGC , containing visceral motor neurons and the hypaxial motor column HMC , innervating the abdominal walls, are present at thoracic levels.

Medial motor column MMC motor neurons, innervating proximal muscles, are present all along the spinal cord. D, dorsal; V, ventral. Dopamine neurons and oculomotor neurons are induced in the ventral neural tube of the midbrain through the actions of the morphogens Sonic hedgehog Shh , fibroblast growth factor 8 Fgf8 , Wnt1 and retinoic acid RA Figure 2. Midbrain dopamine neurons are generated from neuroepithelial cells of the floor plate that have a non-neurogenic character Andersson et al.

The floor plate is a specialized glial structure located in the most ventral midline of the neural tube from the midbrain to the tail region Strahle et al.

It controls neuronal subtype specification along the dorsoventral D-V axis through the secretion of Shh Jessell, The capacity of the floor plate to give rise to neurons is restricted along the rostrocaudal axis of the brain, where floor plate cells located caudally to the midbrain do not normally give rise to neurons Ono et al.

Figure 2. Morphogen signaling during neural tube development. During embryogenesis, neurons present in the midbrain MB , including dopamine and oculomotor neurons are born at the intersection of the signaling molecules Shh, Wnt1, and Fgf8.

Sagital and coronal views at the midbrain and spinal cord levels of the mouse embryo showing the expression patterns of these morphogens. Adapted from Aguila et al. The isthmic organizer forms a boundary between the midbrain and hindbrain and controls patterning of the midbrain and the anterior hindbrain. It is essential for the specification and normal development of dopamine neurons and serotonin neurons Brodski et al.

Several signaling factors, including Shh, Fgf8, Fgf17, Fgf18, and Wnt1, are expressed by and around the isthmic organizer and are involved in this process Figure 2. The combination of Shh and Fgf8 is necessary for the induction of dopamine neurons in the rostral forebrain and the lateral midbrain Wang et al. However, Shh is no longer required after embryonic E day E At this developmental stage, Foxa2, a forkhead transcription factor, induced by Shh, is essential for the generation of midbrain dopamine neurons Ferri et al.

ChIP-seq analysis recently showed that Foxa2 positively regulated determinants of dopamine neurons, including the transcription factors Lmx1a, Lmx1b, Msx1, and Ferd31, while repressing ventrolateral genes in midbrain dopamine progenitors, including Helt, Tle4, Otx1, Sox1, and Tal2. Foxa2 also repressed the expression of Gli1, Gli2, and Gli3, which are the intracellular transducers of Shh signaling Mavromatakis et al.

Foxa2 has previously been shown to regulate the expression of the orphan transcription factor NR4A2 Nurr1 Ferri et al. In addition, Foxa2 appears to play a role in regulating axon trajectories around the midline through targets including Slit2 and 3 Metzakopian et al. The transcription factors Lmx1a and Lmx1b are important for the specification of dopamine neurons Smidt et al.

Importantly, dopamine progenitors in the developing midbrain can be subdivided into molecularly distinct medial and lateral domains and these show differential sensitivity to the loss of Lmx1a and Lmx1b. Here, Lmx1a converts non-neuronal floor plate cells into medial dopamine progenitors Andersson et al. However, Lmx1a is not absolutely required for the generation of these neurons, since the deficiency in dopamine neurogenesis in the Lmx1a knockout mouse recovers over time Deng et al.

Lmx1b is required for generating lateral dopamine progenitors, that in fact do not appear to originate from the floor plate, and which are selectively ablated in Lmx1b mutants Deng et al. Oculomotor neurons are generated immediately lateral to the dopamine progenitor domain from cells that express Lmx1b, Phox2a, Sim1, and Nkx6.

The sequential generation of oculomotor neurons and red nucleus neurons seems controlled by Lmx1b, which is more broadly expressed than Lmx1a in the early developing midbrain, through the activation of Phox2a Deng et al.

Phox2a, in turn, is required and sufficient to promote oculomotor neuron fate Pattyn et al. Sim1, which is co-expressed with Phox2a in this progenitor pool, likely contributes to the specification of red nucleus neurons, but its activity appears suppressed by Phox2a Deng et al.

Furthermore, the migration of newly born oculomotor neurons appears regulated by the Shh-inducible homedomain trancription factor Nkx6. During early development, starting at E9, Fgf8 is expressed by the isthmic organizer Heikinheimo et al.

Fgf8 appears to maintain normal development of the midbrain and hindbrain by regulating transcription factors such as engrailed-1 En1 , En2, and Pax5 Liu et al. Furthermore, Fgf8 appears to regulate anteroposterior patterning of dopamine neurons in a cell-autonomous manner and is neccessary for maturation of dopamine neurons Lahti et al.

Fgf8 also directs the rostral growth of axons from midbrain dopamine neurons by inducing the repulsion factor semaphorin 3F Yamauchi et al. Wnt1 expression precedes Fgf8, starting at E8. During early somite stages, Wnt1 is broadly expressed in the presumptive mesencephalon, but following neural tube closure, the expression gradually becomes refined to a narrow band of cells located immediately rostral to the isthmus and the dorsal midline of the CNS Parr et al.

While Wnt1 does not have isthmic-like activity, it is essential, and deletion results in loss of midbrain and cerebellar structures by E10 and a reduction in the number of midbrain dopamine neurons McMahon and Bradley, ; Thomas and Capecchi, ; McMahon et al.

Wnt1 appears necessary for the development of midbrain dopamine neurons since Fgf8 and Shh fail to induce expression of tyrosine hydroxylase TH the rate limiting enzyme in dopamine synthesis and the homedomain transcription factor Pitx3 which is normally selectively expressed in midbrain dopamine neurons in the Wnt1 knockout mouse Prakash et al.

Wnt1 and Wnt5 double knockout mice showed a greater loss of dopamine progenitors and neurons than single mutants, indicating that Wnt1 and Wnt5a cooperate to promote midbrain dopamine neurogenesis Andersson et al. Ectopic expression of Wnt1 in the posterior hindbrain can induce midbrain dopamine neurons through the activation of the transcription factor Otx2 and the repression of Gbx2 and Nkx2.

If ectopic Wnt signaling is combined with restored Lmx1b levels, midbrain dopamine neurons can be generated at even more posterior levels of the hindbrain, but not in the spinal cord Joksimovic et al.

Interestingly, Otx2 appears to determine the anterior identity that confers neurogenic potential of floor plate cells.

Consequently, ectopic expression of Otx2 in the ventral hindbrain induces midbrain dopamine neurons, partly by inducing Lmx1a from floor plate cells, which normally do not give rise to neurons Ono et al. While Wnt1 expression is largely unaffected by Lmx1a loss-of-function, Lmx1b is a crucial regulator of Wnt1 expression in midbrain dopamine progenitors at later developmental stages Deng et al. In addition to the role of canonical Wnt signaling in early specification, Wnt1 and Wnt3a increase neurogenesis and regulate the proliferation of Nurr1-positive midbrain dopamine precursor cells Castelo-Branco et al.

Likewise, disruption of canonical Wnt signaling leads to neurogenesis defects and perturbs the migration and segregation of midbrain dopamine neurons Tang et al. Wnt2 is also involved in midbrain dopamine neurogenesis through activation of the canonical pathway Sousa et al.

Wnt5a increases the number of midbrain dopamine neurons by promoting the acquisition of a fully mature dopaminergic phenotype through upregulation of Pitx3 expression Castelo-Branco et al.

Wnt5a is also thought to control morphogenesis, dopamine progenitor cell division, cell cycle exit Andersson et al. RA also seems to play a role in midbrain dopamine neuron differentiation.

Retinal dehydrogenase 1 Raldh1 , which converts retinaldehyde into RA, is expressed in the vental midbrain already at E9. Raldh1 and thus the RA level is transcriptionally regulated in the midbrain by Pitx3 Chung et al. Deficiency in Pitx3 results in the selective loss of substantia nigra compacta dopamine neurons Hwang et al.

Maternal supplementation of RA could partially rescue SNc dopamine neuron degeneration in the Pitx3 knockout mice Jacobs et al. Pitx3 appears to regulate genes in both an RA-dependent and an RA-independent manner, which could explain the partial, but not complete, rescue of SNc dopamine neurons after RA supplementation in Pitx3 knockout mice. Other morphogens and growth factors are important for survival and maturation of midbrain dopamine neurons.

GDNF appears to act as a target-derived neurotrophic factor through its high expression in striatal neurons that are innervated by SNc dopamine neurons Hwang et al. In addition, GDNF is transiently expressed in the midbrain during dopamine neuron specification.

Pitx3 is in turn required for activating the expression of brain-derived neurotrophic factor BDNF in a subpopulation of SNc dopamine neurons during embryogenesis. Motor neurons are the main regulators of movement, they extend axons that reach the periphery from the CNS and innervate skeletal and smooth muscles, forming neuromuscular junctions NMJs.

In the fatal neurodegenerative diseases amyotrophic lateral sclerosis ALS and spinal muscular atrophy SMA , motor neurons degenerate, leading to progressive paralysis and respiratory failure. Motor neurons are found at different levels of the CNS, in the cortex, midbrain, hindbrain and spinal cord. Muscle fibers can be intra or extrafusal according to their role in motor control.

Importantly, these motor neuron groups show differential vulnerability to degeneration in ALS Kawamura et al. Motor neurons differ in cell size based on their functional properties and the fiber type they innervate Henneman et al.

For instance, the motor unit formation muscle fibers innervated by the same motor neuron has a hierarchical composition, and the size of the recruited motor neurons is correlated to the muscle unit force.

For this reason, FF and FR motor neurons, that are characterized by higher force requirement, are larger in size than S motor neurons Kernell et al. Slow and fast twitch motor neurons can be distinguished by a set of markers, at least during adulthood.

For example, the expression of the synaptic vesicle protein SV2A is described to be restricted to S motor neurons Chakkalakal et al.

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Oct 26,  ·![Figure][1] Hot seat. Investigations are under way into Hisashi Moriguchi's iPS cell treatment claims. CREDIT: KYODO/AP A startling case involving a groundbreaking stem cell experiment by an unknown researcher, a bogus Harvard affiliation, and multiple collaborators who appear to have signed their names to papers they knew little about is raising questions about the research enterprise Cited by: 1. Aug 20,  · ALS Ice Bucket Challenge Has Raised $ Million In Three Weeks. Kevin Eggan, a faculty member at the Harvard Stem Cell Institute, said that some promising possible treatments ended up not working in clinical trials, however, the money raised for ALS research via the Ice Bucket Challenge could help with treatments to extend the lifespan of Author: Patricia Didelot. Jul 31,  · Skin cells could help with treatment of Alzheimer's Thu 31 Jul "No one has ever managed to isolate these neurons from a patient and grow them in a dish," said Kevin Eggan Author: Ian Sample.