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cAMP-dependent reorganization of the Cajal bodies and splicing machinery in cultured Schwann cells.

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genetics, Animals, drug effects, Up-Regulation, Transcription, Genetic, Survival of Motor Neuron 1 Protein, metabolism, cytology, Schwann Cells, SMN Complex Proteins, S Phase, Ribonucleoproteins, Small Nuclear, Rats, Sprague-Dawley, Rats, RNA-Binding Proteins, RNA Splicing, Protein-Serine-Threonine Kinases, growth & development, Peripheral Nerves, Nerve Tissue Proteins, Histones, Gene Expression Regulation, DNA Replication, Cyclin-Dependent Kinases, Cyclin-Dependent Kinase 2, Cyclic AMP Response Element-Binding Protein, pharmacology, Cyclic AMP, Colforsin, Coiled Bodies, Cells, Cultured, Cell Nucleus, Cell Division, CDC2-CDC28 Kinases, Animals, Newborn

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      Abstract

      It is well established that forskolin-induced elevation of cAMP results in activation of DNA synthesis in Schwann cell cultures. This promitotic response is partially mediated by the Cdk2, which is required for the transition from the G1 to the S phase of the cell cycle. In the present study, we analyze the effects of cAMP elevation in cultured Schwann cells on the transcriptional activity and on the organization of two nuclear compartments involved in pre-mRNA processing: Cajal bodies (CBs) and splicing factor compartments. Our immunofluorescence and quantitative studies show that forskolin treatment induces a 5.6-fold increase in the proportion of S phase Schwann cells, detected by a short pulse (20 min) of BrdU incorporation. This increase in DNA synthesis correlates with an activation of global transcription, as is indicated by the higher nuclear incorporation of BrU in nascent RNA. Forskolin treatment significantly increases the percentage of Schwann cells containing typical CBs, which concentrate spliceosomal snRNPs and the survival motor neuron (SMN) protein. This increase in the number of CBs closely correlates with the activation of transcription. Moreover, the occurrence of CBs is significantly higher in BrdU (+) cells than in BrdU (-) cells, indicating that entry in the S phase promotes the formation of CBs. During the S phase, Schwann cell nuclei display higher Cdk2 nuclear staining and concentrate this kinase in CBs. Forskolin also induces a redistribution of the pre-mRNA splicing factors in Schwann cells. Primary cultures of Schwann cells provide an excellent physiological model to demonstrate that the assembly of CBs is a transcription- and replication-dependent cellular event. Moreover, the S phase accumulation of Cdk2 observed in Schwann cells supports a functional link between CBs and DNA replication, which is mediated by the possible participation of CBs in the regulatory control of histone gene expression. Copyright 2002 Wiley-Liss, Inc.

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      Journal
      10.1002/glia.10157
      12420317

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