Progenitor cells from embryonic chick dorsal root ganglia differentiate in vitro to neurons: biochemical and electrophysiological evidence.

Rohrer, Hermann; Henke-Fahle, Sigrid; El-Sharkawy, T. Y.; Lux, H. D.; Thoenen, H.

doi:10.1002/j.1460-2075.1985.tb03840.x

Cited by 78 publications

(53 citation statements)

References 30 publications

(25 reference statements)

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“…To address the issue of whether Mk is derived from the neuronal or non-neuronal SG population, E7 SG were dissociated and neurons were immunopurified using the neuron-specific Q211 antibody (Rohrer et al, 1985;Rohrer and Thoenen, 1987;Zackenfels et al, 1995 The selective NB development in SG when compared with parasympathetic ganglia suggests that proliferation of parasympathetic neuron progenitors is not regulated by Alk signaling. Indeed, much lower Alk expression levels (8.3±0.1-fold, s.e.m.…”

Section: Forced Expression Of Constitutively Active Alk Mutations Asmentioning

confidence: 99%

Midkine and Alk signaling in sympathetic neuron proliferation and neuroblastoma predisposition

et al. 2011

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SUMMARYNeuroblastoma (NB) is the most common extracranial solid tumor in childhood and arises from cells of the developing sympathoadrenergic lineage. Activating mutations in the gene encoding the ALK tyrosine kinase receptor predispose for NB. Here, we focus on the normal function of Alk signaling in the control of sympathetic neuron proliferation, as well as on the effects of mutant ALK. Forced expression of wild-type ALK and NB-related constitutively active ALK mutants in cultures of proliferating immature sympathetic neurons results in a strong proliferation increase, whereas Alk knockdown and pharmacological inhibition of Alk activity decrease proliferation. Alk activation upregulates NMyc and trkB and maintains Alk expression by an autoregulatory mechanism involving Hand2. The Alk-ligand Midkine (Mk) is expressed in immature sympathetic neurons and in vivo inhibition of Alk signaling by virus-mediated shRNA knockdown of Alk and Mk leads to strongly reduced sympathetic neuron proliferation. Taken together, these results demonstrate that the extent and timing of sympathetic neurogenesis is controlled by Mk/Alk signaling. The predisposition for NB caused by activating ALK mutations may thus be explained by aberrations of normal neurogenesis, i.e. elevated and sustained Alk signaling and increased NMyc expression. Activation of Alk signaling, in particular overexpression of ALK wt and NB ALK mutants resulted in the upregulation of NMyc and trkB, which may contribute to continued proliferation and NB predisposition. MATERIALS AND METHODS Expression plasmidsPcDNA3.1 expression plasmids for human ALK wt , ALK F1174L and ALK R1275Q, and pCAGGS expression plasmids for Hand2, Phox2b wt and Phox2b K155X have been described previously (Janoueix-Lerosey et al., 2008;Reiff et al., 2010). shRNA constructsThe shRNA against Gallus gallus Alk and Mk were designed using BLOCK-iT RNAi Designer (Invitrogen, Karlsruhe, Germany) and target the following sequences: shAlk, 5Ј-AAU GGU UUC UCU CUA UGU CCA ACU C-3Ј; shMk, 5Ј-GAG CUG ACU GCA AGU ACA AGU UUG A-3Ј. Scrambled shRNAs (sc-shRNA, Invitrogen, Karlsruhe, Germany) served as controls. In situ hybridizationNon-radioactive in situ hybridization on cryosections and preparation of digoxigenin labeled probes for chick Phox2b was carried out as described previously (Ernsberger et al., 1997;Stanke et al., 1999). qPCR analysisEqual amounts of RNA were used to synthesize cDNA with Oligo(dT)-primers and Superscript-III-reverse-transcriptase according to manufacturer's instructions (Invitrogen, Karlsruhe, Germany). The PCR was carried out using Abgene's Absolute Blue SYBR-Green qPCR Mix (Abgene, Epsom, UK) in a Stratagene Mx3000p Light Cycler (Stratagene, Waldbronn, Germany). All primers (MWG Biotech AG, Ebersberg, Germany) were designed to anneal optimally at 58°C with PerlPrimer (Marshall, 2004) (95°C for 30 seconds, 58°C for 30 seconds, 72°C for 30 seconds, repeated for 50 cycles and a subsequent dissociation curve). The primer pairs (see Table S1 in the supplementary material) were ...

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Section: Forced Expression Of Constitutively Active Alk Mutations Asmentioning

confidence: 99%

Midkine and Alk signaling in sympathetic neuron proliferation and neuroblastoma predisposition

et al. 2011

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“…Between E4 and E7, chick DRG contain progenitors whose initial survival is independent of NG,F (Ernsberger and Rohrer, 1988). As development proceeds, sensory neuron sur-viva1 can be enhanced by NGF, brain-derived neurotrophic factor (BDNF), and neurotrophin-3 (NT3) (Levi-Montalcini and Angeletti, 1968;Barde et al, 1980;Lindsay et al, 1985;Rohrer et al, 1985;Ernfors et al, 1990;Hohn et al, 1990;Maisonpierre et al, 1990).…”

Section: Stem Cell Factor Is a Neurotrophicmentioning

confidence: 99%

Stem cell factor is a neurotrophic factor for neural crest-derived chick sensory neurons

1994

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We have found that stem cell factor (SCF) selectively enhances the survival of cultured embryonic chick dorsal root ganglia (DRG) neurons. Neurons grown in the presence of SCF expressed both neurofilament 150 kDa subunit and calcitonin-gene related peptide. SCF does not, however, enhance the survival of parasympathetic, placode-derived sensory or sympathetic neurons in culture. Combining SCF with brain-derived neurotrophic factor or neurotrophin-3, but not with NGF, maintains more neurons than either factor alone, suggesting that these factors have partially overlapping activities. SCF preferentially rescues small neurons from the DRG. Labeling studies with bromodeoxyuridine indicate that the neurons sustained by SCF are not differentiating from a dividing progenitor.

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“…At all the stages studied here, ganglia contain indifferent cells (undifferentiated, neural progenitors), primitive neuroblasts, and intermediate neuroblasts (Pannese, 1974). It has been shown that neurons that have only recently developed from precursors are identifiable as SDB cells (Rohrer et al, 1985;Ernsberger and Rohrer, f 988;Wright et al, 1992). Like these authors, we, therefore, assumed that SDB cells were not axotomized by the isolation procedure and that they therefore grew their first neurites in culture.…”

Section: Quantitative Analysis Of Neurite Outgrowth In Culturementioning

confidence: 71%

“…To investigate further the possibility that newly formed neurons may behave differently from others in culture, measurements of neurite length were made from a morphologically distinct population of immature (SDB) neurons. These neurons had not been axotomized; in other words, they extended their first neurites in culture (Rohrer et al, 1985;Ernsberger and Rohrer, 1988;Wright et al, 1492). When these new neurons were taken from different segmental levels and cultured, statistically significant differences in the amounts of neurite outgrowth after 24 hours were observed.…”

Section: Segmental Independence Of Neurite Outgrowthmentioning

confidence: 99%

“…The shape of the distributions could be related to properties of neuronal subpopulations within the DRG, which might behave differently when placed into culture. However, although numerous cell surface and intracellular markers have been used to identify subpopulations of DRG cells (Rohrer et al, 1985(Rohrer et al, , 1986Marusich et al, 1986;New and Mudge, 1986;Sieber-Blum, 1989;Scott et al, 1990), it remains unclear how these are related to functionally distinct subsets within very immature DRG. For instance, mature ganglia contain histochemically distinct populations of small, dorsomedially located neurons, and larger, ventrolaterally located neurons (Hamburger and Levi-Montalchi, 1949;Hamburger et al, 198 1 ).…”

Section: Distribution Of Neurite Lengthsmentioning

confidence: 99%

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Segmental independence and age dependence of neurite outgrowth from embryonic chick sensory neurons

Parson

Ribchester

1995

J. Neurobiol.

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SUMMARYTargets in limb regions of the chick embryo are further removed from the dorsal root ganglia that innervate them compared with thoracic ganglion-to-target distances. It has been inferred that axons grow into the limb regions two to three times faster than into nonlimb regions. We tested whether the differences were due to intrinsic properties of the neurons located at different segmental levels. Dorsal root ganglia (DRG) were isolated from the forelimb, trunk, and hind limb regions of stage 25-30 embryos. Neurite outgrowth was measured in dissociated cell culture and in cultures of DRG explants. Although there was considerable variability in the amount of neurite outgrowth, there were no substantive differences in the amount or the rate of outgrowth comparing brachial, thoracic, or lumbosacral neurons. The amount of neurite outgrowth in dissociated cell cultures increased with the stage of development. Overall, our data suggest that DRG neurons express a basal amount of outgrowth, which is initially independent of target-derived neurotrophic influences; the magnitude of this intrinsic growth potential increases with stage of development; and the neurons of the DRG are not intrinsically specified to grow neurites at rates that are matched to the distance they are required to grow to make contact with their peripheral targets in vivo. We present a speculative model based on Poisson statistics, which attempts to account for the variability in the amount of neurite outgrowth from dissociated neurons.

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Progenitor cells from embryonic chick dorsal root ganglia differentiate in vitro to neurons: biochemical and electrophysiological evidence.

Cited by 78 publications

References 30 publications

Midkine and Alk signaling in sympathetic neuron proliferation and neuroblastoma predisposition

Midkine and Alk signaling in sympathetic neuron proliferation and neuroblastoma predisposition

Stem cell factor is a neurotrophic factor for neural crest-derived chick sensory neurons

Segmental independence and age dependence of neurite outgrowth from embryonic chick sensory neurons

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