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Rapid redistribution of F-actin that precedes the neurite outgrowth of PC12D cells in response to staurosporine and nerve growth factor
5-23
CO-CULTURE
SYSTEM
AUTONOMIC NERVOUS SYSTEM. Institute
of
Technology,
USING
PC-12
YASUHISA ~____._._. Matsugasaki,
CELLS
ENDO,
AS
Department
Sakyo,
kyoto
A MODEL OF THE PERIPHERAL of 606,
Applied
Biology,
Kyoto
Japan
the peripheral autonomic nerves have no clear synaptic contact with their target In general, various endocrine and exocrine cells etc. In order to clarify cells including smooth muscle cells, the innervation mechanism of them, we have tried to prepare an 112 vitro constant model using PC-12 cells as a neuronal element, Ski-3 cells as a model of smooth muscles and several ccl I lines. RINr cells as a mode1 of endocrine pancreas were co-cultured under various conditions. The relationships between the neurites of PC-12 and other cellular elements were investigated by and intracellular microinjection techniques of immunohistochemistry, electron microscopy, exocytotic release of large-cored vesicles of PC-12 was also fluorescent dyes. The non-synaptical, demonstrated ultrastructurally in this co-culture system, under the condition of high K+ and Ca”‘.
5-24
RAPID REDISTRIBUTION OF F-ACTIN THAT PRECEDES THE NEURITE OUTGROWTH OF PC12D CELLS IN RESPONSE TO STAUROSPORINE AND NERVE GROWTH FACTOR. Institute for Developmental Research, Aichi MISAO IWANAGA AND MAMORU SANO, Prefectural Colony, Kasuaai. Aichi 480-03, Jaoan PC12D cells, a subline of ~~12 cells, extend neurites very quickly in response to NGF or to dbcAMP even when RNA synthesis was blocked. A few minutes after the addition of such agents, staining of F-actin revealed the formation of a prominent microfilament network around the periphery of cells where ruffling occurs. We have implicated that the redistribution of F-actin and subsequent outgrowth of neurites in PC12D cells in response to NGF or to dbcAMP may be triggered by the activation of an inhibitor of protein Nevertheless, staurosporine, specific protein kinases. We showed kinase, has been found to induce the outgrowth of neurites in PC12 cells. that this alkaloid indeed caused rapid outgrowth of neurites in PClZD cells and in in PCl2D chick dorsal root ganglion. The bundles of F-actin induced by staurosporine cells were remarkably thick and were different in shape from that induced by NGF or dbcAMP. Cytochalasin B selectively inhibited the outgrowth of neurites in response to NGF or to dbcAMP. to staurosporine but do not significantly that in response However, colchicine only slightly affected the formation of neurites in response to The results suggest the possibility that the staurosporine-induced staurosporine. protrusions may not be complete neurites and may be large filopodia.
CO-CULTURE
SYSTEM
AUTONOMIC NERVOUS SYSTEM. Institute
of
Technology,
USING
PC-12
YASUHISA ~____._._. Matsugasaki,
CELLS
ENDO,
AS
Department
Sakyo,
kyoto
A MODEL OF THE PERIPHERAL of 606,
Applied
Biology,
Kyoto
Japan
the peripheral autonomic nerves have no clear synaptic contact with their target In general, various endocrine and exocrine cells etc. In order to clarify cells including smooth muscle cells, the innervation mechanism of them, we have tried to prepare an 112 vitro constant model using PC-12 cells as a neuronal element, Ski-3 cells as a model of smooth muscles and several ccl I lines. RINr cells as a mode1 of endocrine pancreas were co-cultured under various conditions. The relationships between the neurites of PC-12 and other cellular elements were investigated by and intracellular microinjection techniques of immunohistochemistry, electron microscopy, exocytotic release of large-cored vesicles of PC-12 was also fluorescent dyes. The non-synaptical, demonstrated ultrastructurally in this co-culture system, under the condition of high K+ and Ca”‘.
5-24
RAPID REDISTRIBUTION OF F-ACTIN THAT PRECEDES THE NEURITE OUTGROWTH OF PC12D CELLS IN RESPONSE TO STAUROSPORINE AND NERVE GROWTH FACTOR. Institute for Developmental Research, Aichi MISAO IWANAGA AND MAMORU SANO, Prefectural Colony, Kasuaai. Aichi 480-03, Jaoan PC12D cells, a subline of ~~12 cells, extend neurites very quickly in response to NGF or to dbcAMP even when RNA synthesis was blocked. A few minutes after the addition of such agents, staining of F-actin revealed the formation of a prominent microfilament network around the periphery of cells where ruffling occurs. We have implicated that the redistribution of F-actin and subsequent outgrowth of neurites in PC12D cells in response to NGF or to dbcAMP may be triggered by the activation of an inhibitor of protein Nevertheless, staurosporine, specific protein kinases. We showed kinase, has been found to induce the outgrowth of neurites in PC12 cells. that this alkaloid indeed caused rapid outgrowth of neurites in PClZD cells and in in PCl2D chick dorsal root ganglion. The bundles of F-actin induced by staurosporine cells were remarkably thick and were different in shape from that induced by NGF or dbcAMP. Cytochalasin B selectively inhibited the outgrowth of neurites in response to NGF or to dbcAMP. to staurosporine but do not significantly that in response However, colchicine only slightly affected the formation of neurites in response to The results suggest the possibility that the staurosporine-induced staurosporine. protrusions may not be complete neurites and may be large filopodia.