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Identification of the Cytolinker Protein Plectin in Neuronal Cellst-Expression of a Rodless Isoform in Neurons of the Rat Superior Cervical Ganglion

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Summary

Plectin, a large (≤500 kDa) dumbbell-shaped cytolinker protein plays an important role in the organization of the cytoskeletal network and the maintenance of cell integrity in a wide varietyof tissues and cell types. Earlier experiments revealed the presence of plectin in the central nervous system, whereas the expression in the peripheral nervous system remained unclear. Our results obtained with reverse transcriptase-PCR (RT-PCR) provide evidence that plectin is expressed in structures of the rat peripheral nervous system. In addition to well-characterized plectin transcriptswewere able to reveal novel splicing variants affecting the region coding for the central rod domain. Previous studies report a high, but tissuespecific variability of the N-terminal domain of plectin due to alternatively spliced first coding exons and the optionally spliced small exons 2á and 3á. We demonstrate for the first time, using single-cell RT-PCR and immunocytochemistry, that plectin is expressed in neurons of the rat superior cervical ganglion (SCG). Plectin transcripts of single SCG neurons, starting with exon 1c as the first coding exon, contain the optionally spliced exon 2α but lack exon 31. These data therefore suggest that plectin is expressed in rat SCG neurons as a rodless isoform with the molecular mass of 390 kDa.

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Abbreviations

ABD:

actin-binding domain

BPAG:

bullous pemphigoid antigen

bp:

base pairs

cDNA:

complementaryDNA

DRG:

dorsal root ganglion

EBS:

epidermolysis bullosa simplex

GAPDH:

glyceraldehyde-3-phosphate dehydrogenase

IF:

intermediate filament

mAb:

monoclonal antibody

MAP:

microtubule associated protein

MD:

muscular dystrophy

NF:

neurofilament

NMJ:

neuromuscular junction

PBS:

phosphate buffered saline

PSD:

postsynaptic density

RT-PCR:

reverse transcriptase-PCR

SCG:

superior cervical ganglion

References

  • Andrae, K., Lassmann, H., Bittner, R., Shorny, S., Fassler, R., Propst, F., and Wiche, G. (1997). Targeted inactivation of plectin reveals essential function in maintaining the integrity of skin, muscle, and heart cytoarchitecture. Genes Dev. 11:3143–3156.

    Article  Google Scholar 

  • Andrae, K., Nikolic, B., Stocher, M., Drenckhahn, D., and Wiche, G. (1998). Not just scaffolding: Plectin regulates actin dynamics in cultured cells. Genes Dev. 12:3442–3451.

    Article  Google Scholar 

  • Boehm, S., Huck, S. (1995). Alpha 2-adrenoreceptor-mediated inhibition of acetylcholine-induced noradrenaline release from rat sympathetic neurons: An action at voltage-gated Ca2+ channels. Neuroscience 69:221–231.

    Article  CAS  PubMed  Google Scholar 

  • Brown, A., Bernier, G., Mathieu, M., Rossant, J., and Kothary, R. (1995). The mouse dystonia musculorum gene is a neural isoform of bullous pemphigoid antigen 1. Nat. Genet. 10:301–306.

    Article  CAS  PubMed  Google Scholar 

  • Chomczynski, P., Sacchi, N. (1987). Single-step method of RNA isolation by acid guanidinium thiocyanate–phenol–chloroform extraction. Anal. Biochem. 162:156–159.

    Article  CAS  PubMed  Google Scholar 

  • Clubb, B. H., Chou, Y. H., Herrmann, H., Svitkina, T. M., Borisy, G. G., and Goldman, R. D. (2000). The 300-kDa intermediate filament-associated protein (IFAP300) is a hamster plectin ortholog. Biochem. Biophys. Res. Commun. 273:183–187.

    Article  CAS  PubMed  Google Scholar 

  • Dowling, J., Yang, Y., Wollmann, R., Reichardt, L. F., and Fuchs, E., (1997). Developmental expression of BPAG1-n: Insights into the spastic ataxia and gross neurologic degeneration in dystonia musculorum mice. Dev. Biol. 187:131–142.

    Article  CAS  PubMed  Google Scholar 

  • Dowling, J., Yu, Q. C., Fuchs, E. (1996). Beta4 integrin is required for hemidesmosome formation, cell adhesion and cell survival. J. Cell Biol. 134:559–572.

    Article  CAS  PubMed  Google Scholar 

  • Eger, A., Stockinger, A., Wiche, G., and Foisner, R. (1997). Polarisation-dependent association of plectin with desmoplakin and the lateral submembrane skeleton in MDCK cells. J. Cell Sci. 110:1307–1316.

    CAS  PubMed  Google Scholar 

  • Elliott, C. E., Becker, B., Oehler, S., Castanon, M. J., Hauptmann, R., and Wiche, G. (1997). Plectin transcript diversity: Identification and tissue distribution of variants with distinct first coding exons and rodless isoforms. Genomics 42:115–125.

    Article  CAS  PubMed  Google Scholar 

  • Errante, L. D., Wiche, G., and Shaw, G. (1994). Distribution of plectin, an intermediate filament-associated protein, in the adult rat central nervous system. J. Neurosci. Res. 37:515–528.

    Article  CAS  PubMed  Google Scholar 

  • Foisner, R., Feldman, B., Sander, L., Seifert, G., Artlieb, U., and Wiche, G. (1994). A panel of monoclonal antibodies to rat plectin: Distinction by epitope mapping and immunoreactivity with different tissues and cell lines. Acta Histochem. 96:421–438.

    CAS  PubMed  Google Scholar 

  • Foisner, R., Feldman, B., Sander, L., and Wiche, G. (1991). Monoclonal antibody mapping of structural and functional plectin epitopes. J. Cell Biol. 112:397–405.

    Article  CAS  PubMed  Google Scholar 

  • Foisner, R., Leichtfried, F. E., Herrmann, H., Small, J. V., Lawson, D., and Wiche, G. (1988). Cytoskeleton-associated plectin: In situ localization, in vitro reconstitution, and binding to immobilized intermediate filament proteins. J. Cell Biol. 106:723–733.

    Article  CAS  PubMed  Google Scholar 

  • Foisner, R., and Wiche, G. (1987). Structure and hydrodynamic properties of plectin molecules. J. Mol. Biol. 198:515–531.

    Article  CAS  PubMed  Google Scholar 

  • Fontao, L., Geerts, D., Kuikman, I., Koster, J., Kramer, D., and Sonnenberg, A. (2001). The interaction of plectin with actin: Evidence for cross-linking of actin filaments by dimerization of the actin-binding domain of plectin. J. Cell Sci. 114:2065–2076.

    CAS  PubMed  Google Scholar 

  • Fuchs, E., and Cleveland, D. W. (1998). A structural scaffolding of intermediate filaments in health and disease. Science 279:514–519.

    Article  CAS  PubMed  Google Scholar 

  • Fuchs, P., Zörer, M., Rezniczek, G. A., Spazierer, D., Oehler, S., Castanon, M. J., Hauptmann, R., and Wiche, G. (1999). Unusual 5′ transcript complexity of plectin isoforms: Novel tissue-specific exons modulate actin binding activity. Hum. Mol. Genet. 8:2461–2472.

    Article  CAS  PubMed  Google Scholar 

  • Gache, Y., Chavanas, S., Lacour, J. P., Wiche, G., Owaribe, K., Meneguzzi, G., and Ortonne, J. P. (1996). Defective expression of plectin/HD1 in epidermolysis bullosa simplex with muscular dystrophy. J. Clin. Invest. 97:2289–2298.

    Article  CAS  PubMed  Google Scholar 

  • Gao, F. B., Brenman, J. E., Jan, L. Y., and Jan, Y. N. (1999). Genes regulating dendritic outgrowth, branching, and routing in Drosophila. Genes Dev. 13:2549–2561.

    Article  CAS  PubMed  Google Scholar 

  • Gregory, S. L., and Brown, N. H. (1998). Kakapo, a gene required for adhesion between and within cell layers in Drosophila, encodes a large cytoskeletal linker protein related to plectin and dystrophin. J. Cell Biol. 143:1271–1282.

    Article  CAS  PubMed  Google Scholar 

  • Guo, L., Degenstein, L., Dowling, J., Yu, Q. C., Wollmann, R., Perman, B., and Fuchs, E. (1995). Gene targeting of BPAG1: Abnormalities in mechanical strength and cell migration in stratified epithelia and neurologic degeneration. Cell 81:233–243.

    Article  CAS  PubMed  Google Scholar 

  • Herrmann, H., and Wiche, G. (1987). Plectin and IFAP-300K are homologous proteins binding to microtubule-associated proteins 1 and 2 and to the 240-kilodalton subunit of spectrin. J. Biol. Chem. 262:1320–1325.

    CAS  PubMed  Google Scholar 

  • House, C. M., Frew, I. J., Huang, H. L., Wiche, G., Traficante, N., Nice, E., Catimel, B., and Bowtell, D. D. (2003). Proc. Natl. Acad. Sci., U.S.A. 100:3101–3106.

    Article  CAS  PubMed  Google Scholar 

  • Kletter, G., Evans, O. B., Lee, J. A., Melvin, B., Yates, A. B., and Bock, H. G. O. (1989). Congenital muscular dystrophy and epidermolysis bullosa simplex. J. Pediatr. 114:105–106.

    Google Scholar 

  • Leung, C. L., Green, K. J., and Liem, R. K. (2002). Plakins: A family of versatile cytolinker proteins. Trends Cell Biol. 12:37–45.

    Article  CAS  PubMed  Google Scholar 

  • Leung, C. L., Zheng, M., Prater, S. M., and Liem, R. K. (2001). The BPAG1 locus: Alternative splicing produces multiple isoforms with distinct cytoskeletal linker domains, including predominant isoforms in neurons and muscles. J. Cell Biol. 154:691–697.

    Article  CAS  PubMed  Google Scholar 

  • Liu, C. G., Maercker, C., Castanon, M. J., Hauptmann, R., and Wiche, G. (1996). Human plectin: Organization of the gene, sequence analysis, and chromosome localization (8q24). Proc. Natl. Acad. Sci. U.S.A. 93:4278–4283.

    Article  CAS  PubMed  Google Scholar 

  • Lunter, P. C., and Wiche, G. (2002). Direct binding of plectin to Fer kinase and negative regulation of its catalytic activity. Biochem. Biophys. Res.Commun. 296:904–910.

    Article  CAS  PubMed  Google Scholar 

  • McLean, W. H., Pulkkinen, L., Smith, F. J., Rugg, E. L., Lane, E. B., Bullrich, F., Burgeson, R. E., Amano, S., Hudson, D. L., Owaribe, K., McGrath, J. A., McMillan, J. R., Eady, R. A., Leigh, I. M., Christiano, A. M., and Uitto, J. (1996). Loss of plectin causes epidermolysis bullosa with muscular dystrophy: cDNA cloning and genomic organization. Genes Dev. 10:1724–1735.

    Article  CAS  PubMed  Google Scholar 

  • Nikolic, B., Mac Nulty, E., Mir, B., and Wiche, G. (1996). Basic amino acid residue cluster within nuclear targeting sequence motif is essential for cytoplasmic plectin–vimentin network junctions. J. Cell Biol. 134:1455–1467.

    Article  CAS  PubMed  Google Scholar 

  • Osmanagic-Myers, S., and Wiche, G. (2004). Plectin-RACK1 (receptor for activated C kinase 1) scaffolding: A novel mechanism to regulate protein kinase C activity. J. Biol. Chem. 279:18701–18710.

    Article  CAS  PubMed  Google Scholar 

  • Prokop, A., Uhler, J., Roote, J., and Bate, M. (1998). The kakapo mutation affects terminal arborization and central dendritic sprouting of Drosophila motorneurons. J. Cell Biol. 143:1283–1294.

    Article  CAS  PubMed  Google Scholar 

  • Pytela, R., and Wiche, G. (1980). High molecular weight polypeptides (270,000–340,000) from cultured cells are related to hog brain microtubule-associated proteins but copurify with intermediate filaments. Proc. Natl. Acad. Sci. U.S.A. 77:4808–4812.

    Article  CAS  PubMed  Google Scholar 

  • Reipert, S., Steinböck, F., Fischer, I., Bittner, R. E., Zeold, A., and Wiche, G. (1999). Association of mitochondria with plectin and desmin intermediate filaments in striated muscle. Exp. Cell Res. 252:479–491.

    Article  CAS  PubMed  Google Scholar 

  • Rezniczek, G. A., Abrahamsberg, C., Fuchs, P., Spazierer, D., and Wiche, G. (2003). Plectin 5′-transcript diversity: Short alternative sequences determine stability of gene products, initiation of translation and subcellular localization of isoforms. Hum. Mol. Genet. 12:3181–3194

    Article  CAS  PubMed  Google Scholar 

  • Rezniczek, G. A., de Pereda, J. M., Reipert, S., and Wiche, G. (1998). Linking integrin alpha6beta4-based cell adhesion to the intermediate filament cytoskeleton: Direct interaction between the beta4 subunit and plectin at multiple molecular sites. J. Cell Biol. 141:209–225.

    Article  CAS  PubMed  Google Scholar 

  • Ruhrberg, C., Hajibagheri, M. A., Parry, D. A., and Watt, F. M. (1997). Periplakin, a novel component of cornified envelopes and desmosomes that belongs to the plakin family and forms complexes with envoplakin. J. Cell Biol. 139:1835–1849.

    Article  CAS  PubMed  Google Scholar 

  • Smith, F. J. D., Eady, R. A. J., Leigh, I. M., McMillan, J. R., Rugg, E. L., Kelsell, D. P., Bryant, S. P., Spurr, N. K., Geddes, J. F., Kirtschig, G., Milana, G., de Bono, A. G., Owaribe, K., Wiche, G., Pulkkinen, L., Uitto, J., McLean, W. H. I., and Lane, E. B. (1996). Plectin deficiency results in muscular dystrophy with epidermolysis bullosa. Nat. Genet. 13:450–457

    Article  CAS  PubMed  Google Scholar 

  • Steinböck, F. A., Nikolic, B., Coulombe, P. A., Fuchs, E., Traub, P., and Wiche, G. (2000). Dose-dependent linkage, assembly inhibition and disassembly of vimentin and cytokeratin 5/14 filaments through plectin's intermediate filament-binding domain. J. Cell Sci. 113:483–491.

    PubMed  Google Scholar 

  • Steinböck, F. A., and Wiche, G. (1999). Plectin: A cytolinker by design. Biol. Chem. 380:151–158.

    Article  PubMed  Google Scholar 

  • Wiche, G. (1998). Role of plectin in cytoskeleton organization and dynamics. J. Cell Sci. 111:2477–2486.

    CAS  PubMed  Google Scholar 

  • Wiche, G., Becker, B., Luber, K., Weitzer, G., Castanon, M. J., Hauptmann, R., Stratowa, C., and Stewart, M. (1991). Cloning and sequencing of rat plectin indicates a 466-kD polypeptide chain with a three-domain structure based on a central alpha-helical coiled coil. J. Cell Biol. 114:83–99.

    Article  CAS  PubMed  Google Scholar 

  • Yang, Y., Dowling, J., Yu, Q. C., Kouklis, P., Cleveland, D. W., and Fuchs, E. (1996). An essential cytoskeletal linker protein connecting actin microfilaments to intermediate filaments. Cell 86:655–665.

    Article  CAS  PubMed  Google Scholar 

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Authors and Affiliations

  1. Institute ofCancerResearch,Department ofMedicine I, MedicalUniversity of Vienna, Vienna, Austria

    Ferdinand Steinboeck

  2. Division of Biochemistry and Molecular Biology, Brain Research Institute, Medical University of Vienna, Vienna, Austria

    Doris Kristufek

  3. Institute of Cancer Research, Department of Medicine I, Medical University of Vienna, Borschkegasse 8a, A-1090, Vienna, Austria

    Ferdinand Steinboeck

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  1. Ferdinand Steinboeck
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  2. Doris Kristufek
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Correspondence to Ferdinand Steinboeck.

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Steinboeck, F., Kristufek, D. Identification of the Cytolinker Protein Plectin in Neuronal Cellst-Expression of a Rodless Isoform in Neurons of the Rat Superior Cervical Ganglion. Cell Mol Neurobiol 25, 1151–1169 (2005). https://doi.org/10.1007/s10571-005-8503-0

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