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In vitro differentiation of neural crest cells into sensory-like neurons
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SUBPOP~TIONS OF NEURAL CREST CELLS AND NON-NEURONAL DORSAL ROOT GANGLION CELLS EXPRESS A SENSORY NEURON SPECIFIC EPITOPE. M.F. Marusich, K. Pourmehr, and J.A. Weston. Dept. of Biology, University of Oregon, Eugene, OR 97403. We have produced a monoclonal antibody (SNI) that binds a cell surface epitope expressed on a subpopulation of avian sensory neurons. Detectable levels of the antibody do not bind to other neurons or non-neurons within the PNS or CNS, with the exception of a small population of cells lacking neuronal morphology within sensory and sympathetic ganglia. The SNI(+), non-neuronal cells are found in sensory ganglia both prior to and after the appearance of SNI(+) neurons. We have also observed that SNI(+), morphologically non-neuronal cells arise in cultures of neural crest. It is possible therefore, that the SNI(+), non-neuronal cells observed in sensory and sympathetic ganglia represent neural crest derived cells at some intermediate stage of neurogenesis. Supported by NSF Grant PCM-8218899, NIH Grant DE-04316 and NIH Postdoctoral Fellowahip 5 F32 HD 06292 to M.M.
IN VITRO DIFFERENTIATION OF NEURAL CREST CELLS INTO SENSORY-LIKE NEURONS. M. Sieber-Blum and S. Patel. The Johns Hopkins University, Baltimore, MD 21205. In vivo neural crest progeny give rise to autonomic and sensory neurons. However, only terminally differentiated autonomic neurons have been described in culture. We here report the in vitro differentiation of quail neural crest cells into substance P- and VlP-like immunoreactive neurons. These neurons were bipolar or pseudounipolar. Processes lacked varicosities. Cell bodies were large (20 ~) and rounded. Addition of fibronectin and NGF caused an increase in the number of neurons per culture. Autonomic-like neurons that stained with antibodies against substance P and VIP were also observed. They were smaller (8-10 ~) and multipolar, and their processes had numerous varicosities. Large cells stained for carbonic anhydrase, which is concentrated in sensory neurons. Small cells were negative. These newly detected neurons may prove useful for the in vitro clonal analysis of neuronal lineages. (Supported by NIH grant HDI5311 and by a grant from the Dysautonomia Foundation)
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ORIGIN OF THE THYMIC MYOID CELLS IN BIRDS. H. Nakamura, K. Watanabe, C. Ayer-Le Lievre, and N. Le Douarin. Dept. of Anatomy, Hiroshima Univ. Sch. of Med., Kasumi 1-2-3, Minami-ku, Hiroshima 734, Japan, Dept. of Anatomy, Fukui Med. Sch., Fukui, Japan and Institut d'Embryologie du CNRS et du Coll~ge de France, Nogent-sur-Marne, France We have attempted to determine the origin of thymic myoid cells in birds by transplanting quail neural crest into a chick embryo. Thymuses were removed from operated embryos just before hatching, when myoid cells first appear. After staining with antibodies specific to muscle creatine kinase, sections were stained by the Feulgen reaction to identify quail nuclei. Other samples were processed for electron microscopy. Both methods demonstrated that myoid cells had quail nuclei and therefore one can conclude that thymic myoid cells in birds originate from the neural crest. (Supported by Grants No. 83-01-33 from NCNMMD of the Ministry of Health and Welfare, and No. 59213011 for Special Project Research from the Ministry of Education Sciences and Culture, Japan.)
REVERSAL OF A DEVELOPMENTAL RESTRICTION IN NEURAL CREST-DERIVED CELLS OF AVIAN EMBRYOS BY THE CO-CARCINOGEN, 12-0TETRADECANOYLPHORBOL-13-ACETATE (TPA). G. Ciment and J.A. Weston, Department of Biology, University of Oregon, Eugene, Oregon 97403. The neural crest of early vertebrate embryos gives rise to a variety of derivative tissues, including pigment cells. Earlier studies of Nichols and Weston (1977) showed that 5 day embryonic chick dorsal root ganglia--another crest derivative--contain cells which undergo adventitious pigmentation in culture, suggesting that the developmental restriction of melanogenesis in crest cells occurs after this developmental stage. We have reexamined this phenomenon using the co-carcinogenic drug TPA. We have found that this drug dramatically extends the developmental period in which adventitious pigmentation in dorsal root ganglion cells can be produced. This drug effect is dose- and stage-dependent and also occurs with other crest derivatives. We interpret these results to mean that TPA can reverse this restriction of melanogenesis occuring during early crest development. NSF grant PCM8218899.
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SUBPOP~TIONS OF NEURAL CREST CELLS AND NON-NEURONAL DORSAL ROOT GANGLION CELLS EXPRESS A SENSORY NEURON SPECIFIC EPITOPE. M.F. Marusich, K. Pourmehr, and J.A. Weston. Dept. of Biology, University of Oregon, Eugene, OR 97403. We have produced a monoclonal antibody (SNI) that binds a cell surface epitope expressed on a subpopulation of avian sensory neurons. Detectable levels of the antibody do not bind to other neurons or non-neurons within the PNS or CNS, with the exception of a small population of cells lacking neuronal morphology within sensory and sympathetic ganglia. The SNI(+), non-neuronal cells are found in sensory ganglia both prior to and after the appearance of SNI(+) neurons. We have also observed that SNI(+), morphologically non-neuronal cells arise in cultures of neural crest. It is possible therefore, that the SNI(+), non-neuronal cells observed in sensory and sympathetic ganglia represent neural crest derived cells at some intermediate stage of neurogenesis. Supported by NSF Grant PCM-8218899, NIH Grant DE-04316 and NIH Postdoctoral Fellowahip 5 F32 HD 06292 to M.M.
IN VITRO DIFFERENTIATION OF NEURAL CREST CELLS INTO SENSORY-LIKE NEURONS. M. Sieber-Blum and S. Patel. The Johns Hopkins University, Baltimore, MD 21205. In vivo neural crest progeny give rise to autonomic and sensory neurons. However, only terminally differentiated autonomic neurons have been described in culture. We here report the in vitro differentiation of quail neural crest cells into substance P- and VlP-like immunoreactive neurons. These neurons were bipolar or pseudounipolar. Processes lacked varicosities. Cell bodies were large (20 ~) and rounded. Addition of fibronectin and NGF caused an increase in the number of neurons per culture. Autonomic-like neurons that stained with antibodies against substance P and VIP were also observed. They were smaller (8-10 ~) and multipolar, and their processes had numerous varicosities. Large cells stained for carbonic anhydrase, which is concentrated in sensory neurons. Small cells were negative. These newly detected neurons may prove useful for the in vitro clonal analysis of neuronal lineages. (Supported by NIH grant HDI5311 and by a grant from the Dysautonomia Foundation)
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ORIGIN OF THE THYMIC MYOID CELLS IN BIRDS. H. Nakamura, K. Watanabe, C. Ayer-Le Lievre, and N. Le Douarin. Dept. of Anatomy, Hiroshima Univ. Sch. of Med., Kasumi 1-2-3, Minami-ku, Hiroshima 734, Japan, Dept. of Anatomy, Fukui Med. Sch., Fukui, Japan and Institut d'Embryologie du CNRS et du Coll~ge de France, Nogent-sur-Marne, France We have attempted to determine the origin of thymic myoid cells in birds by transplanting quail neural crest into a chick embryo. Thymuses were removed from operated embryos just before hatching, when myoid cells first appear. After staining with antibodies specific to muscle creatine kinase, sections were stained by the Feulgen reaction to identify quail nuclei. Other samples were processed for electron microscopy. Both methods demonstrated that myoid cells had quail nuclei and therefore one can conclude that thymic myoid cells in birds originate from the neural crest. (Supported by Grants No. 83-01-33 from NCNMMD of the Ministry of Health and Welfare, and No. 59213011 for Special Project Research from the Ministry of Education Sciences and Culture, Japan.)
REVERSAL OF A DEVELOPMENTAL RESTRICTION IN NEURAL CREST-DERIVED CELLS OF AVIAN EMBRYOS BY THE CO-CARCINOGEN, 12-0TETRADECANOYLPHORBOL-13-ACETATE (TPA). G. Ciment and J.A. Weston, Department of Biology, University of Oregon, Eugene, Oregon 97403. The neural crest of early vertebrate embryos gives rise to a variety of derivative tissues, including pigment cells. Earlier studies of Nichols and Weston (1977) showed that 5 day embryonic chick dorsal root ganglia--another crest derivative--contain cells which undergo adventitious pigmentation in culture, suggesting that the developmental restriction of melanogenesis in crest cells occurs after this developmental stage. We have reexamined this phenomenon using the co-carcinogenic drug TPA. We have found that this drug dramatically extends the developmental period in which adventitious pigmentation in dorsal root ganglion cells can be produced. This drug effect is dose- and stage-dependent and also occurs with other crest derivatives. We interpret these results to mean that TPA can reverse this restriction of melanogenesis occuring during early crest development. NSF grant PCM8218899.
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