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Rae Nishi - One of the best experts on this subject based on the ideXlab platform.
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neuronal differentiation from postmitotic precursors in the Ciliary Ganglion
Developmental Biology, 2002Co-Authors: John Sechrist, Vivian M Lee, Marianne Bronnerfraser, Rae NishiAbstract:In the chick Ciliary Ganglion, neuronal number is kept constant between St. 29 and St. 34 (E6–E8) despite a large amount of cell death. Here, we characterize the source of neurogenic cells in the Ganglion as undifferentiated neural crest-derived cells. At St. 29, neurons and nonneuronal cells in the Ciliary Ganglion expressed the neural crest markers HNK-1 and p75^(NTR). Over 50% of the cells were neurons at St. 29; of the nonneuronal cells, a small population expressed glial markers, whereas the majority was undifferentiated. When placed in culture, nonneuronal cells acquired immunoreactivity for HuD, suggesting that they had commenced neuronal differentiation. The newly differentiated neurons arose from precursors that did not incorporate bromodeoxyuridine. To test whether these precursors could undergo neural differentiation in vivo, purified nonneuronal cells from St. 29 quail ganglia were transplanted into chick embryos at St. 9–14. Subsequently, quail cells expressing neuronal markers were found in the chick Ciliary Ganglion. The existence of this precursor pool was transient because nonneuronal cells isolated from St. 38 ganglia failed to form neurons. Since all Ciliary Ganglion neurons are born prior to St. 29, these results demonstrate that there are postmitotic neural crest-derived precursors in the developing Ciliary Ganglion that can differentiate into neurons in the appropriate environment.
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cell death and neuronal replacement during formation of the avian Ciliary Ganglion
Developmental Biology, 2001Co-Authors: Vivian M Lee, Gregory G Smiley, Rae NishiAbstract:Programmed cell death is a prominent feature of embryonic development and is essential in matching the number of neurons to the target tissues that are innervated. Although a decrease in neuronal number which coincides with peripheral synaptogenesis has been well documented in the avian Ciliary Ganglion, it has not been clear whether cell death also occurs earlier. We observed TUNEL-positive neurons as early as stage 24, with a large peak at stage 29. This cell death at stage 29 was followed by a statistically significant (P < 0.0001) decrease in total neuron number at stage 31. The total number of neurons was recovered by stage 33/34. This suggested that dying neurons were replaced by new neurons. This replacement process did not involve proliferation because bromodeoxyuridine applied at stages 29 and 31 was unable to label neurons harvested at stage 33/34. The peak of cell death at stage 29 was increased 2.3-fold by removal of the optic vesicle and was reduced by 50% when chCNTF was overexpressed. Taken together, these results suggest that the regulation of neuron number in the Ciliary Ganglion is a dynamic process involving both cell death and neural replacement from postmitotic precursors prior to differentiation and innervation of target tissues.
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activin a and follistatin influence expression of somatostatin in the Ciliary Ganglion in vivo
Developmental Biology, 1998Co-Authors: Diane C Darland, Rae NishiAbstract:An important developmental question concerns whether neurotransmitter phenotype is an inherent property of neurons or is influenced by target tissues. This issue can be addressed in the avian Ciliary Ganglion (CG) which contains two cholinergic populations, Ciliary and choroid neurons, that differentially express the peptide cotransmitter, somatostatin. The present study tests the hypothesis that differences in the level of expression of activin A and its endogenous inhibitor follistatin in CG neuron target tissues are responsible for selective expression of somatostatin in choroid neurons. Intraocular injection of activin A or follistatin (300 ng injected at E10/E11) in cultured embryos resulted in a 39% increase or a 23% decrease, respectively, in somatostatin-positive neurons relative to controls. Chorioallantoic membrane application of follistatin (1 mg daily from E7 to E13) reduced somatostatin positive neurons by 54%. Neuron number, size, and target tissue morphology were unaffected by these treatments. Together with our previous studies, these data suggest that activin A and follistatin are target-derived molecules that regulate neuropeptide phenotype in the Ciliary Ganglion. © 1998 Academic Press
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overexpression of Ciliary neurotrophic factor in vivo rescues chick Ciliary Ganglion neurons from cell death
Journal of Neurobiology, 1998Co-Authors: Tom P Finn, Songte Kim, Rae NishiAbstract:Ciliary Ganglion (CG) neurons undergo target-dependent cell death during embryonic development. Although Ciliary neurotrophic factor (CNTF) was identified in vitro by its ability to support the survival of chick CG neurons, its function as a target-derived neurotrophic factor has been questioned by those working on mammalian-derived forms of CNTF. We have purified and cloned a chicken CNTF [chCNTF; formerly growth-promoting activity (GPA)] that is expressed in CG targets during the period of cell death and is secreted by cells transfected with chCNTF. In the present study we used a retroviral vector, RCASBP(A), to overexpress chCNTF in CG target tissues. Elevation of chCNTF biological activity three- to fourfold in the embryonic eye rescued an average of 31% of the neurons that would have normally died in vivo. In some individuals, nearly all of the neurons were rescued. ChCNTF had no effect on the number of neurons observed prior to cell death, nor were there any deleterious effects of either viral infection or overexpression of CNTF. These results show that chCNTF is able to function in vivo as a trophic factor for CG neurons, and suggest that limited availability of trophic support is one of the factors regulating CG neuron survival during development.
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activin a and follistatin expression in developing targets of Ciliary Ganglion neurons suggests a role in regulating neurotransmitter phenotype
Neuron, 1995Co-Authors: Diane C Darland, Brian A Link, Rae NishiAbstract:Abstract The avian Ciliary Ganglion contains choroid neurons that innervate choroid vasculature and express somatostatin as well as Ciliary neurons that innervate iris/Ciliary body but do not express somatostatin. We have previously shown in culture that activin A induces somatostatin immunoreactivity in both neuron populations. We now show in vivo that both targets contain activin A; however, choroid expressed higher levels of activin A mRNA. In contrast, follistatin, an activin A inhibitor, was higher in iris/Ciliary body. Iris cell-conditioned medium also contained an activity that inhibited activin A and could be depleted with antifollistatin antibodies. These results suggest that development of somatostatin is limited to choroid neurons by differential expression of activin A and follistatin in Ciliary Ganglion targets.
Kerstin Krieglstein - One of the best experts on this subject based on the ideXlab platform.
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Data_Sheet_2_Bid Expression Network Controls Neuronal Cell Fate During Avian Ciliary Ganglion Development.ZIP
2018Co-Authors: Sophie Koszinowski, Hauke Busch, Kerstin Krieglstein, Veronica La Padula, Frank Edlich, Melanie BoerriesAbstract:Avian Ciliary Ganglion (CG) development involves a transient execution phase of apoptosis controlling the final number of neurons, but the time-dependent molecular mechanisms for neuronal cell fate are largely unknown. To elucidate the molecular networks regulating important aspects of parasympathetic neuronal development, a genome-wide expression analysis was performed during multiple stages of avian CG development between embryonic days E6 and E14. The transcriptome data showed a well-defined sequence of events, starting from neuronal migration via neuronal fate cell determination, synaptic transmission, and regulation of synaptic plasticity to growth factor associated signaling. In particular, we extracted a neuronal apoptosis network that characterized the cell death execution phase at E8/E9 and apoptotic cell clearance at E14 by combining the gene time series analysis with network synthesis from the chicken interactome. Network analysis identified TP53 as key regulator and predicted involvement of the BH3 interacting domain death agonist (BID). A virus-based RNAi knockdown approach in vivo showed a crucial impact of BID expression on the execution of ontogenetic programmed cell death (PCD). In contrast, Bcl-XL expression did not impact PCD. Therefore, BID-mediated apoptosis represents a novel cue essential for timing within CG maturation.
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Data_Sheet_1_Bid Expression Network Controls Neuronal Cell Fate During Avian Ciliary Ganglion Development.PDF
2018Co-Authors: Sophie Koszinowski, Hauke Busch, Kerstin Krieglstein, Veronica La Padula, Frank Edlich, Melanie BoerriesAbstract:Avian Ciliary Ganglion (CG) development involves a transient execution phase of apoptosis controlling the final number of neurons, but the time-dependent molecular mechanisms for neuronal cell fate are largely unknown. To elucidate the molecular networks regulating important aspects of parasympathetic neuronal development, a genome-wide expression analysis was performed during multiple stages of avian CG development between embryonic days E6 and E14. The transcriptome data showed a well-defined sequence of events, starting from neuronal migration via neuronal fate cell determination, synaptic transmission, and regulation of synaptic plasticity to growth factor associated signaling. In particular, we extracted a neuronal apoptosis network that characterized the cell death execution phase at E8/E9 and apoptotic cell clearance at E14 by combining the gene time series analysis with network synthesis from the chicken interactome. Network analysis identified TP53 as key regulator and predicted involvement of the BH3 interacting domain death agonist (BID). A virus-based RNAi knockdown approach in vivo showed a crucial impact of BID expression on the execution of ontogenetic programmed cell death (PCD). In contrast, Bcl-XL expression did not impact PCD. Therefore, BID-mediated apoptosis represents a novel cue essential for timing within CG maturation.
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Table_1_Bid Expression Network Controls Neuronal Cell Fate During Avian Ciliary Ganglion Development.XLSX
2018Co-Authors: Sophie Koszinowski, Hauke Busch, Kerstin Krieglstein, Veronica La Padula, Frank Edlich, Melanie BoerriesAbstract:Avian Ciliary Ganglion (CG) development involves a transient execution phase of apoptosis controlling the final number of neurons, but the time-dependent molecular mechanisms for neuronal cell fate are largely unknown. To elucidate the molecular networks regulating important aspects of parasympathetic neuronal development, a genome-wide expression analysis was performed during multiple stages of avian CG development between embryonic days E6 and E14. The transcriptome data showed a well-defined sequence of events, starting from neuronal migration via neuronal fate cell determination, synaptic transmission, and regulation of synaptic plasticity to growth factor associated signaling. In particular, we extracted a neuronal apoptosis network that characterized the cell death execution phase at E8/E9 and apoptotic cell clearance at E14 by combining the gene time series analysis with network synthesis from the chicken interactome. Network analysis identified TP53 as key regulator and predicted involvement of the BH3 interacting domain death agonist (BID). A virus-based RNAi knockdown approach in vivo showed a crucial impact of BID expression on the execution of ontogenetic programmed cell death (PCD). In contrast, Bcl-XL expression did not impact PCD. Therefore, BID-mediated apoptosis represents a novel cue essential for timing within CG maturation.
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Bid Expression Network Controls Neuronal Cell Fate During Avian Ciliary Ganglion Development
Frontiers Media S.A., 2018Co-Authors: Sophie Koszinowski, Hauke Busch, Kerstin Krieglstein, Veronica La Padula, Frank Edlich, Melanie BoerriesAbstract:Avian Ciliary Ganglion (CG) development involves a transient execution phase of apoptosis controlling the final number of neurons, but the time-dependent molecular mechanisms for neuronal cell fate are largely unknown. To elucidate the molecular networks regulating important aspects of parasympathetic neuronal development, a genome-wide expression analysis was performed during multiple stages of avian CG development between embryonic days E6 and E14. The transcriptome data showed a well-defined sequence of events, starting from neuronal migration via neuronal fate cell determination, synaptic transmission, and regulation of synaptic plasticity to growth factor associated signaling. In particular, we extracted a neuronal apoptosis network that characterized the cell death execution phase at E8/E9 and apoptotic cell clearance at E14 by combining the gene time series analysis with network synthesis from the chicken interactome. Network analysis identified TP53 as key regulator and predicted involvement of the BH3 interacting domain death agonist (BID). A virus-based RNAi knockdown approach in vivo showed a crucial impact of BID expression on the execution of ontogenetic programmed cell death (PCD). In contrast, Bcl-XL expression did not impact PCD. Therefore, BID-mediated apoptosis represents a novel cue essential for timing within CG maturation
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RARβ regulates neuronal cell death and differentiation in the avian Ciliary Ganglion
Developmental Neurobiology, 2015Co-Authors: Sophie Koszinowski, Melanie Boerries, Hauke Busch, Kerstin KrieglsteinAbstract:Programmed cell death during chicken Ciliary Ganglion (CG) development is mostly discussed as an extrinsically regulated process, guided either by the establishment of a functional balance between preGanglionic and postGanglionic activity or the availability of target-derived neurotrophic factors. We found that the expression of the gene coding for the nuclear retinoic acid receptor β (RARB) is transiently upregulated prior to and during the execution phase of cell death in the CG. Using retroviral vectors, the expression of RARB was knocked down during embryonic development in ovo. The knockdown led to a significant increase in CG neuron number after the cell death phase. BrdU injections and active caspase-3 staining revealed that this increase in neuron number was due to an inhibition of apoptosis during the normal cell death phase. Furthermore, apoptotic neuron numbers were significantly increased at a stage when cell death is normally completed. While the cholinergic phenotype of the neurons remained unchanged after RARB knockdown, the expression of the proneural gene Cash1 was increased, but somatostatin-like immunoreactivity, a hallmark of the mature choroid neuron population, was decreased. Taken together, these results point toward a delay in neuronal differentiation as well as cell death. The availability of nuclear retinoic acid receptor β (RARβ) and RARβ-induced transcription of genes could therefore be a new intrinsic cue for the maturation of CG neurons and their predisposition to undergo cell death. © 2015 Wiley Periodicals, Inc. Develop Neurobiol 75: 1204–1218, 2015
Vivian M Lee - One of the best experts on this subject based on the ideXlab platform.
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both neural crest and placode contribute to the Ciliary Ganglion and oculomotor nerve
Developmental Biology, 2003Co-Authors: Vivian M Lee, John Sechrist, Simone Luetolf, Marianne BronnerfraserAbstract:The chick Ciliary Ganglion is a neural crest-derived parasympathetic Ganglion that innervates the eye. Here, we examine its axial level of origin and developmental relationship to other ganglia and nerves of the head. Using small, focal injections of DiI, we show that neural crest cells arising from both the caudal half of the midbrain and the rostral hindbrain contribute to the Ciliary as well as the trigeminal Ganglion. Precursors to both ganglia have overlapping migration patterns, moving first ventrolaterally and then rostrally toward the optic vesicle. At the level of the midbrain/forebrain junction, precursors to the Ciliary Ganglion separate from the main migratory stream, turn ventromedially, and condense in the vicinity of the rostral aorta and Rathke's pouch. Ciliary neuroblasts first exit the cell cycle at early E2, prior to and during Ganglionic condensation, and neurogenesis continues through E5.5. By E3, markers of neuronal differentiation begin to appear in this population. By labeling the ectoderm with DiI, we discovered a new placode, caudal to the eye and possibly contiguous to the trigeminal placode, that contributes a few early differentiating neurons to the Ciliary Ganglion, oculomotor nerve, and connecting branches to the ophthalmic nerve. These results suggest for the first time a dual neural crest and placodal contribution to the Ciliary Ganglion and associated nerves.
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neuronal differentiation from postmitotic precursors in the Ciliary Ganglion
Developmental Biology, 2002Co-Authors: John Sechrist, Vivian M Lee, Marianne Bronnerfraser, Rae NishiAbstract:In the chick Ciliary Ganglion, neuronal number is kept constant between St. 29 and St. 34 (E6–E8) despite a large amount of cell death. Here, we characterize the source of neurogenic cells in the Ganglion as undifferentiated neural crest-derived cells. At St. 29, neurons and nonneuronal cells in the Ciliary Ganglion expressed the neural crest markers HNK-1 and p75^(NTR). Over 50% of the cells were neurons at St. 29; of the nonneuronal cells, a small population expressed glial markers, whereas the majority was undifferentiated. When placed in culture, nonneuronal cells acquired immunoreactivity for HuD, suggesting that they had commenced neuronal differentiation. The newly differentiated neurons arose from precursors that did not incorporate bromodeoxyuridine. To test whether these precursors could undergo neural differentiation in vivo, purified nonneuronal cells from St. 29 quail ganglia were transplanted into chick embryos at St. 9–14. Subsequently, quail cells expressing neuronal markers were found in the chick Ciliary Ganglion. The existence of this precursor pool was transient because nonneuronal cells isolated from St. 38 ganglia failed to form neurons. Since all Ciliary Ganglion neurons are born prior to St. 29, these results demonstrate that there are postmitotic neural crest-derived precursors in the developing Ciliary Ganglion that can differentiate into neurons in the appropriate environment.
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cell death and neuronal replacement during formation of the avian Ciliary Ganglion
Developmental Biology, 2001Co-Authors: Vivian M Lee, Gregory G Smiley, Rae NishiAbstract:Programmed cell death is a prominent feature of embryonic development and is essential in matching the number of neurons to the target tissues that are innervated. Although a decrease in neuronal number which coincides with peripheral synaptogenesis has been well documented in the avian Ciliary Ganglion, it has not been clear whether cell death also occurs earlier. We observed TUNEL-positive neurons as early as stage 24, with a large peak at stage 29. This cell death at stage 29 was followed by a statistically significant (P < 0.0001) decrease in total neuron number at stage 31. The total number of neurons was recovered by stage 33/34. This suggested that dying neurons were replaced by new neurons. This replacement process did not involve proliferation because bromodeoxyuridine applied at stages 29 and 31 was unable to label neurons harvested at stage 33/34. The peak of cell death at stage 29 was increased 2.3-fold by removal of the optic vesicle and was reduced by 50% when chCNTF was overexpressed. Taken together, these results suggest that the regulation of neuron number in the Ciliary Ganglion is a dynamic process involving both cell death and neural replacement from postmitotic precursors prior to differentiation and innervation of target tissues.
Michele H. Jacob - One of the best experts on this subject based on the ideXlab platform.
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dystrophin and utrophin isoforms are expressed in glia but not neurons of the avian parasympathetic Ciliary Ganglion
Brain Research, 2008Co-Authors: Rachel C Blitzblau, Elizabeth K Storer, Michele H. JacobAbstract:Muscular dystrophy patients often show cognitive impairment, in addition to muscle degeneration caused by dystrophin gene defects. The cognitive impairments lead to speculation that the dystrophin protein family may play a key role at neuronal synapses. Dystrophin regulates the stability of selected GABAA receptor subtypes and α3-containing nicotinic acetylcholine receptors (nAChRs) at a subset of central GABAergic and peripheral sympathetic nicotinic neuron synapses. Similarly, utrophin, the autosomal homologue of dystrophin, is not required for clustering but indirectly stabilizes muscle-type nAChRs at the neuromuscular junction. We examined dystrophin and utrophin expression and localization in the avian parasympathetic Ciliary Ganglion (CG) to determine whether these proteins play a general role at neuronal nicotinic synapses. We have determined that full-length utrophin and dystrophin and the short dystrophin isoform Dp116 are the major isoforms expressed in the CG based on immunoblotting and immunolabeling. Unexpectedly, the cytoskeletal proteins were not detected at nicotinic synapses or in CG neurons. They are expressed in myelinating and non-myelinating Schwann cells. Further, utrophin expression developmentally precedes that of dystrophin. The proteins show partially overlapping distributions, but also differential accumulation along the surface membrane of Schwann cells adjacent to neuronal somata versus axonal processes. Our findings are consistent with reports that dystrophin protein family members function in the maintenance of cell–cell interactions and myelination by anchoring the Schwann cell surface membrane to the basal lamina. In contrast, our results differ from those in skeletal muscle and a subset of sympathetic neurons where utrophin and dystrophin localize at nicotinic synapses.
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Target Tissues and Innervation Regulate the Characteristics of K+ Currents in Chick Ciliary Ganglion Neurons Developing in situ
The Journal of Neuroscience, 1994Co-Authors: Michelle M. Dourado, Mary E. Wisgirda, Craig L. Brumwell, Michele H. Jacob, Stuart E. DryerAbstract:The expression of appropriate ensembles of ionic channels is necessary for the differentiation and normal function of vertebrate neurons. Cell- cell interactions may regulate the expression and properties of ionic channels in embryonic neurons. Previous studies have shown that the expression of A-type K+ channels (IA) and Ca2+-activated K+ channels (lK[Ca]) is abnormal in chick Ciliary Ganglion neurons developing in vitro in the absence of normal cell-cell interactions. Other voltage- activated currents develop normally under these conditions. The present studies were designed to establish the role of the target tissues and the preGanglionic innervation in regulating the expression of these currents in embryonic chick Ciliary Ganglion neurons developing in situ. Surgical manipulations were used to remove the developing optic vesicle, which contains the target tissues, the mid-dorsal region of the midbrain primordium, which contains the preGanglionic nucleus, or both, all prior to the formation of the Ciliary Ganglion. IA and IK[Ca] were then examined in acutely isolated neurons that developed in ovo in the presence (OV+) or absence (OV-) of the normal target tissues, in the presence (MB+) or absence (MB-) of preGanglionic innervation, and in the absence of both preGanglionic innervation and target tissues (OV- /MB-). The amplitude of IA was unaffected by the operations. However, the activation and inactivation kinetics of IA were two- to threefold faster in OV- or OV-/MB- cells compared to neurons isolated from control OV+ ganglia at embryonic days 11–14 (E11-E14). There were no changes in the voltage dependence of activation or steady-state inactivation, or in the time course of recovery from inactivation. By contrast, neurons isolated from MB- ganglia expressed an IA with amplitude, voltage dependence, and kinetics that were indistinguishable from those of control MB+ and OV+ ganglia. Therefore, interactions with target tissues in the eye play a role in determining the characteristics of IA in developing Ciliary Ganglion neurons, whereas preGanglionic innervation does not. Furthermore, the amplitude of IK[Ca] was reduced by 90–100% in OV-, MB-, and OV-/MB- neurons isolated at E12-E14 as compared to MB+ and OV+ controls. Voltage-activated Ca2+ currents were present at normal amplitudes in all of these neurons. Thus, the expression of IK[Ca] in chick Ciliary Ganglion neurons is regulated by both target tissue interactions and preGanglionic innervation. Therefore, cell-cell interactions are necessary for the expression of a normal ensemble of ionic channels in chick Ciliary Ganglion neurons developing in situ.
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reduced levels of acetylcholine receptor expression in chick Ciliary Ganglion neurons developing in the absence of innervation
The Journal of Neuroscience, 1993Co-Authors: Lynn S Arenella, Jeanne M Oliva, Michele H. JacobAbstract:Chick Ciliary Ganglion neurons receive innervation from a single source, the accessory oculomotor nucleus (AON), and nicotinic ACh receptors (AChRs) mediate chemical synaptic transmission through the Ganglion. Previous experiments examining the developmental expression of AChRs in embryonic chick Ciliary Ganglion neurons in situ have shown that AChR levels increase substantially in the neurons at the time of innervation. Prior to synapse formation, few AChRs are detected in the neurons. In the present experiments, the role of presynaptic inputs in inducing an increase in AChRs was established by examining AChR levels in Ciliary Ganglion neurons that have been deprived of innervation by surgical ablation of the AON prior to synapse formation. AChR levels were dramatically reduced in neurons of input-deprived ganglia as compared to control innervated neurons at all developmental stages examined from embryonic day (ED) 5 to ED 12 as determined by indirect immunocytochemical labeling of frozen Ganglion sections with the anti- AChR monoclonal antibody mAb 35, and light microscopy. In contrast, neuronal somata of input-deprived and control ganglia had equivalent levels of immunolabeling for three other components, a transmembrane glycoprotein of synaptic vesicles, SV2, and two microtubule-associated proteins, MAP 1B and MAP 2, from ED 5 up to ED 10. The results demonstrate that presynaptic inputs specifically increase the levels of AChR expression in developing neurons. In addition, changes in the levels of immunolabeling for AChRs, SV2, MAP 1B, and MAP 2 in neuronal somata after ED 10 demonstrate that other major developmental events also influence the levels of these components in neurons. Declines in the intensity of AChR, SV2, MAP 1B, and MAP 2 immunolabeling within a subset of neuronal somata in both operated and control ganglia at ED 10 and 12 coincide with the period of neuronal cell death. Increases in AChR labeling in the rest of the neuronal population of input-deprived ganglia at ED 12 suggest that, in addition to innervation, synapse formation with the peripheral target tissue influences AChR levels in developing neurons in situ.
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acetylcholine receptor expression in developing chick Ciliary Ganglion neurons
The Journal of Neuroscience, 1991Co-Authors: Michele H. JacobAbstract:Little is known about the levels of nicotinic ACh receptors (AChRs) in neurons prior to innervation and whether the distribution and number of receptors change in response to innervation. In the present study, AChR levels were examined in developing chick Ciliary Ganglion neurons in situ at stages preceding and during early and late phases of synaptogenesis. AChRs were localized in surface and intracellular pools of intact and saponin-permeabilized Ganglionic neurons, respectively, by using a highly sensitive immunocytochemical approach that included the binding of an anti-AChR monoclonal antibody (mAb) followed by a biotinylated secondary antibody and an avidin-biotinylated HRP complex. At older stages of development, embryonic day (ED) 7-7.5 and ED 11, when all of the neurons are known to be receiving synaptic contacts, AChRs were present in both internal and surface pools. Within the neurons, AChRs were associated with organelles that function in the biosynthesis, processing, and transport of integral plasma membrane proteins. On the surface of the neurons, AChRs were predominantly localized in the specialized postsynaptic membrane, with low levels of AChRs being present in extrasynaptic regions. The earliest stage at which synapses could be detected in the Ganglion was ED 4.5. Synapses were detected by light microscopic immunocytochemical labeling with anti-SV2, an mAb to a synaptic vesicle protein, and by ultrastructural analysis. At this stage, most of the neurons were not labeled by the anti-AChR mAb, while a few neurons had dense deposits of reaction product on the rough endoplasmic reticulum and portions of the nuclear envelope. Low levels of reaction product were also found on the surface of a small number of neurons, being localized predominantly on the specialized postsynaptic membrane of the few immature synapses present. Occasionally, small patches of labeling were observed in extrasynaptic regions. In contrast, little internal and no surface anti-AChR immunolabeling was detected in Ciliary Ganglion neurons prior to innervation, at ED 3.5-4. The finding of a large increase in both internal and surface AChR levels in the neurons at the time of innervation suggests that signals from the presynaptic input play an important role in the induction of AChR expression in neurons.
Sophie Koszinowski - One of the best experts on this subject based on the ideXlab platform.
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Data_Sheet_2_Bid Expression Network Controls Neuronal Cell Fate During Avian Ciliary Ganglion Development.ZIP
2018Co-Authors: Sophie Koszinowski, Hauke Busch, Kerstin Krieglstein, Veronica La Padula, Frank Edlich, Melanie BoerriesAbstract:Avian Ciliary Ganglion (CG) development involves a transient execution phase of apoptosis controlling the final number of neurons, but the time-dependent molecular mechanisms for neuronal cell fate are largely unknown. To elucidate the molecular networks regulating important aspects of parasympathetic neuronal development, a genome-wide expression analysis was performed during multiple stages of avian CG development between embryonic days E6 and E14. The transcriptome data showed a well-defined sequence of events, starting from neuronal migration via neuronal fate cell determination, synaptic transmission, and regulation of synaptic plasticity to growth factor associated signaling. In particular, we extracted a neuronal apoptosis network that characterized the cell death execution phase at E8/E9 and apoptotic cell clearance at E14 by combining the gene time series analysis with network synthesis from the chicken interactome. Network analysis identified TP53 as key regulator and predicted involvement of the BH3 interacting domain death agonist (BID). A virus-based RNAi knockdown approach in vivo showed a crucial impact of BID expression on the execution of ontogenetic programmed cell death (PCD). In contrast, Bcl-XL expression did not impact PCD. Therefore, BID-mediated apoptosis represents a novel cue essential for timing within CG maturation.
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Data_Sheet_1_Bid Expression Network Controls Neuronal Cell Fate During Avian Ciliary Ganglion Development.PDF
2018Co-Authors: Sophie Koszinowski, Hauke Busch, Kerstin Krieglstein, Veronica La Padula, Frank Edlich, Melanie BoerriesAbstract:Avian Ciliary Ganglion (CG) development involves a transient execution phase of apoptosis controlling the final number of neurons, but the time-dependent molecular mechanisms for neuronal cell fate are largely unknown. To elucidate the molecular networks regulating important aspects of parasympathetic neuronal development, a genome-wide expression analysis was performed during multiple stages of avian CG development between embryonic days E6 and E14. The transcriptome data showed a well-defined sequence of events, starting from neuronal migration via neuronal fate cell determination, synaptic transmission, and regulation of synaptic plasticity to growth factor associated signaling. In particular, we extracted a neuronal apoptosis network that characterized the cell death execution phase at E8/E9 and apoptotic cell clearance at E14 by combining the gene time series analysis with network synthesis from the chicken interactome. Network analysis identified TP53 as key regulator and predicted involvement of the BH3 interacting domain death agonist (BID). A virus-based RNAi knockdown approach in vivo showed a crucial impact of BID expression on the execution of ontogenetic programmed cell death (PCD). In contrast, Bcl-XL expression did not impact PCD. Therefore, BID-mediated apoptosis represents a novel cue essential for timing within CG maturation.
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Table_1_Bid Expression Network Controls Neuronal Cell Fate During Avian Ciliary Ganglion Development.XLSX
2018Co-Authors: Sophie Koszinowski, Hauke Busch, Kerstin Krieglstein, Veronica La Padula, Frank Edlich, Melanie BoerriesAbstract:Avian Ciliary Ganglion (CG) development involves a transient execution phase of apoptosis controlling the final number of neurons, but the time-dependent molecular mechanisms for neuronal cell fate are largely unknown. To elucidate the molecular networks regulating important aspects of parasympathetic neuronal development, a genome-wide expression analysis was performed during multiple stages of avian CG development between embryonic days E6 and E14. The transcriptome data showed a well-defined sequence of events, starting from neuronal migration via neuronal fate cell determination, synaptic transmission, and regulation of synaptic plasticity to growth factor associated signaling. In particular, we extracted a neuronal apoptosis network that characterized the cell death execution phase at E8/E9 and apoptotic cell clearance at E14 by combining the gene time series analysis with network synthesis from the chicken interactome. Network analysis identified TP53 as key regulator and predicted involvement of the BH3 interacting domain death agonist (BID). A virus-based RNAi knockdown approach in vivo showed a crucial impact of BID expression on the execution of ontogenetic programmed cell death (PCD). In contrast, Bcl-XL expression did not impact PCD. Therefore, BID-mediated apoptosis represents a novel cue essential for timing within CG maturation.
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Bid Expression Network Controls Neuronal Cell Fate During Avian Ciliary Ganglion Development
Frontiers Media S.A., 2018Co-Authors: Sophie Koszinowski, Hauke Busch, Kerstin Krieglstein, Veronica La Padula, Frank Edlich, Melanie BoerriesAbstract:Avian Ciliary Ganglion (CG) development involves a transient execution phase of apoptosis controlling the final number of neurons, but the time-dependent molecular mechanisms for neuronal cell fate are largely unknown. To elucidate the molecular networks regulating important aspects of parasympathetic neuronal development, a genome-wide expression analysis was performed during multiple stages of avian CG development between embryonic days E6 and E14. The transcriptome data showed a well-defined sequence of events, starting from neuronal migration via neuronal fate cell determination, synaptic transmission, and regulation of synaptic plasticity to growth factor associated signaling. In particular, we extracted a neuronal apoptosis network that characterized the cell death execution phase at E8/E9 and apoptotic cell clearance at E14 by combining the gene time series analysis with network synthesis from the chicken interactome. Network analysis identified TP53 as key regulator and predicted involvement of the BH3 interacting domain death agonist (BID). A virus-based RNAi knockdown approach in vivo showed a crucial impact of BID expression on the execution of ontogenetic programmed cell death (PCD). In contrast, Bcl-XL expression did not impact PCD. Therefore, BID-mediated apoptosis represents a novel cue essential for timing within CG maturation
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RARβ regulates neuronal cell death and differentiation in the avian Ciliary Ganglion
Developmental Neurobiology, 2015Co-Authors: Sophie Koszinowski, Melanie Boerries, Hauke Busch, Kerstin KrieglsteinAbstract:Programmed cell death during chicken Ciliary Ganglion (CG) development is mostly discussed as an extrinsically regulated process, guided either by the establishment of a functional balance between preGanglionic and postGanglionic activity or the availability of target-derived neurotrophic factors. We found that the expression of the gene coding for the nuclear retinoic acid receptor β (RARB) is transiently upregulated prior to and during the execution phase of cell death in the CG. Using retroviral vectors, the expression of RARB was knocked down during embryonic development in ovo. The knockdown led to a significant increase in CG neuron number after the cell death phase. BrdU injections and active caspase-3 staining revealed that this increase in neuron number was due to an inhibition of apoptosis during the normal cell death phase. Furthermore, apoptotic neuron numbers were significantly increased at a stage when cell death is normally completed. While the cholinergic phenotype of the neurons remained unchanged after RARB knockdown, the expression of the proneural gene Cash1 was increased, but somatostatin-like immunoreactivity, a hallmark of the mature choroid neuron population, was decreased. Taken together, these results point toward a delay in neuronal differentiation as well as cell death. The availability of nuclear retinoic acid receptor β (RARβ) and RARβ-induced transcription of genes could therefore be a new intrinsic cue for the maturation of CG neurons and their predisposition to undergo cell death. © 2015 Wiley Periodicals, Inc. Develop Neurobiol 75: 1204–1218, 2015