ABSTRACT
Ca(2+)/calmodulin-dependent protein kinase II (CaMK-II) has been linked to the induction of differentiation in preneuronal cells. In these cells, delta isozymes represent the majority of CaMK-IIs expressed and are activated by differentiation stimuli. To determine whether delta CaMK-IIs are causative or coincident with in vitro differentiation, we overexpressed wild-type, constitutively active, and C-terminal domains of delta and gamma CaMK-II isozymes in mouse P19 and NIH/3T3 cells using high-efficiency transfections. At 1-2 days after transfection, only constitutively active delta CaMK-II isozymes induced branched cellular extensions in both cell types. In P19 cells, retinoic acid induced neurite extensions after 3-4 days; these extensions were coincident with a fourfold increase in endogenous CaMK-II activity. Extensions induced by both retinoic acid and delta CaMK-IIs contained class III beta-tubulin in a discontinuous or beaded pattern. C-terminal CaMK-II constructs disrupted the ability of endogenous CaMK-II to autophosphorylate and blocked retinoic acid-induced differentiation. delta CaMK-II was found along extensions, whereas gamma CaMK-II exhibited a more diffuse, cytosolic localization. These data not only support an extranuclear role for CaMK-II in promoting neurite outgrowth, but also demonstrate CaMK-II isozyme specificity in these early steps of neuronal differentiation.
Subject(s)
Calcium-Calmodulin-Dependent Protein Kinases/metabolism , Neoplastic Stem Cells/enzymology , Neurites/enzymology , Neurons/enzymology , 3T3 Cells , Animals , Calcium-Calmodulin-Dependent Protein Kinase Type 2 , Calcium-Calmodulin-Dependent Protein Kinases/genetics , Calcium-Calmodulin-Dependent Protein Kinases/pharmacology , Cell Differentiation/drug effects , Cell Differentiation/genetics , Cell Division/drug effects , Cell Division/genetics , Embryonal Carcinoma Stem Cells , Enzyme Activation/drug effects , Gene Expression , Isoenzymes/genetics , Isoenzymes/metabolism , Mice , Neoplastic Stem Cells/cytology , Neoplastic Stem Cells/drug effects , Neurites/drug effects , Neurons/cytology , Neurons/drug effects , Transfection , Tretinoin/pharmacology , Tumor Cells, CulturedSubject(s)
Antibody Formation/drug effects , Dental Pulp/immunology , Immunity, Cellular/drug effects , Root Canal Filling Materials/pharmacology , Animals , Antigens/immunology , Dental Pulp/drug effects , Dental Pulp Cavity/drug effects , Dental Pulp Cavity/immunology , Dogs , Humans , Lymphocyte Activation/drug effectsABSTRACT
Pulp tissue from three experimental dogs was tested for its antigenicity before and after incubation with CMCP. Reaction to Arthus skin tests showed an increased response to pulp that had been incubated in CMCP, washed, then injected, as compared with pulp or CMCP alone. Hemagglutinating antibody titers (1:360 to 1:450 dilutions) were obtained from pulp tissue altered by CMCP. Inhibition tests showed the specificity of the antibody to be at a dilution titer of 1:350. Therefore, CMCP altered dog pulp tissue and rendered it antigenically active, and a specific humoral response was produced.
Subject(s)
Antibody Formation , Antigens/immunology , Camphor/immunology , Chlorophenols/immunology , Dental Pulp/immunology , Animals , Antibodies/analysis , Dental Pulp Cavity/immunology , Dental Pulp Necrosis/immunology , Dogs , Drug Combinations/immunology , Freund's Adjuvant/administration & dosage , Immunization , Male , Skin TestsABSTRACT
Pulp tissue from three experimental dogs was tested for its antigenicity before and after incubation with AH-26 sealer. identical to Reactions to Arthus skin tests showed an increased response to pulp that had been incubated in AH-26, washed, and then injected, as compared with the material alone. Hemagglutinating antibody titers (1/500 to 1/800) were obtained from pulp tissue altered by AH-26. Inhibition tests showed the apecificity of the antibody to be at a dilution titer of 1:400. Therefore, a mixture of AH-26 sealer altered dog pulp tissue and rendered it antigenically active, and a specific humoral response was produced.
Subject(s)
Antibody Formation , Antigens , Dental Pulp Cavity/immunology , Dental Pulp/immunology , Root Canal Filling Materials , Agglutination Tests , Animals , Antibodies/analysis , Arthus Reaction/immunology , Dental Pulp Necrosis/immunology , Dogs , Immunization , Male , Skin TestsABSTRACT
Pulp tissue from experimental dogs was tested for its antigenicity before and after incubation with Grossman's Formula Sealer (GS). Reactions to Arthus skin tests showed an increased response to pulp that had been incubated in GS, washed, and then injected, as compared with the material alone. Hemagglutinating antibody titers (1:160 to 1:360) were obtained from pulp tissue altered by GS. Inhibition tests showed the specificity of the antibody to be at a dilution titer of 1:200. Therefore, GS altered dog pulp tissue and rendered it antigenically active, and a specific humoral response was produced.
Subject(s)
Antibody Formation , Antigens , Dental Pulp Cavity/immunology , Dental Pulp/immunology , Root Canal Filling Materials/pharmacology , Animals , Antibodies/analysis , Antigens/administration & dosage , Arthus Reaction/immunology , Dogs , Freund's Adjuvant/administration & dosage , Immunization , Male , Skin TestsABSTRACT
Pulp tissue from three experimental dogs was tested for its antigenicity before and after incubation with Kerr (Rickert's) sealer (KS). Reactions to Arthus skin tests showed an increased response to pulp that had been incubated in KS, washed, and then injected, as compared with the material alone. Hemagglutinating antibody titers (1/360--1/600) were obtained from pulp tissue altered by KS. Inhibition tests showed the specificity of the antibody to be at a dilution of titer of 1:300. Therefore, a mixture of Kerr (Rickert's) sealer altered dog pulp tissue and rendered it antigenically active, and a specific humoral response was produced.
