Summary
Small nuclear ribonucleoprotein particles (snRNPs) are subunits of splicing complexes, which show a transcriptiondependent localization pattern. We have analyzed the labelling pattern of snRNPs during pollen development and microspore and pollen embryogenesis inBrassica napus with an antibody which recognizes protein D of Ul, U2, U4, U5, and U6 snRNPs. It was found that nuclei were labelled almost uniformly for snRNPs in microspores and young bicellular pollen. In the generative nuclei of late-bicellular pollen and in the vegetative nuclei and sperm nuclei of mature pollen no snRNPs could be detected. The snRNP-positive nuclei contained mostly one or two brightly labelled nuclear bodies, most likely coiled bodies, often closely related to the nucleolus. These nuclear bodies increased in size from 0.5 um in nuclei of young microspores up to 2 urn in nuclei of late microspores and the vegetative nucleus of early-bicellular pollen. Also their number increased during these developmental stages. After induction of embryogenesis the size of the coiled bodies decreased to about 0.5 μm and in several occasions the coiled body was found free in the nucleoplasm, away from the nucleolus. The results support the idea that the size and number of coiled bodies coincide with changes in general nuclear activity. They also indicate that, in nuclei ofBrassica napus, at least assembly and disassembly of coiled bodies takes place in the nucleoplasm, whereas mature coiled bodies are located adjacent to the nucleolus.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
Abbreviations
- snRNA:
-
small nuclear RNA
- snRNPs:
-
small nuclear ribonucleoprotein particles
References
Alliegro MC, Alliegro MA (1998) Protein heterogeneity in the coiled body compartment. Exp Cell Res 239: 60–68
Andrade LEC, Tan EM, Chan EKL (1993) Immunocytochemical analysis of the coiled body in the cell cycle and during prolifera-tion. Proc Natl Acad Sci USA 90:1947–1951
Beven AF, Simpson GG, Brown JWS, Shaw PJ (1995) The organization of spliceosomal components in the nuclei of higher plants. J Cell Sci 108: 509–518
Billings PB, Allen RW, Jensen FC, Hoch SO (1982) Anti-RNP monoclonal antibodies derived from mouse strain with lupus-like autoimmunity. J Immunol 128:1176–1180
— Barton JR, Hoch SO (1985) A murine monoclonal antibody recognizes the 13,000 molecular weight polypeptide of the Sm small nuclear ribonucleoprotein complex. J Immunol 135: 428–432
Binarova P, Straatman K, Hause B, Hause G, van Lammeren AAM (1993) Nuclear DNA synthesis during the induction of embryo-genesis in cultured microspores and pollen ofBrassica napus L. Theor Appl Genet 87: 9–16
Boudonck K, Dolan L, Shaw PJ (1998) Coiled body numbers in theArabidopsis root epidermis are regulated by cell type, developmental stage and cell cycle parameters. J Cell Sci 111: 3687–3694
——— (1999) The movement of coiled bodies visualized in living plant cells by the green fluorescent protein. Mol Biol Cell 10: 2297–2307
Brasch K, Ochs RL (1992) Nuclear bodies (NB): a newly“rediscovered” organelle. Exp Cell Res 202: 211–223
Carmo-Fonseca M, Pepperkok R, Carvalho MT, Lamond AI (1992) Transcription-dependent co-localization of the Ul, U2, U4/U6 and U5 snRNPs in coiled bodies. J Cell Biol 117:1–14
— Ferreira J, Lamond AI (1993) Assembly of snRNP-containing coiled bodies is regulated in interphase and mitosis: evidence that the coiled body is a kinetic nuclear structure. J Cell Biol 120: 841–852
Christensen AH, Sharrock RA, Quail PA (1992) Maize polyubiqui-tin genes: structure, thermal perturbation of expression and transcript splicing, and promoter activity following transfer to protoplasts by electroporation. Plant Mol Biol 18: 675–689
Concha II, Mansilla J, Riveros M, Burzio LO (1995) Ul snRNP com-ponents are present in the vegetative and generative nuclei of the pollen grain. Sex Plant Reprod 8: 339–344
Custers JBM, Cordewener JHG, Nöllen Y, Dons HJM, van Lookeren Campagne MM (1994) Temperature controls both gameto-phytic and sporophytic development in microspore cultures ofBrassica napus. Plant Cell Rep 13: 267–271
Fakan S (1994) Perichromatin fibrils are in situ forms of nascent transcripts. Trends Cell Biol 4: 86–96
Ferreira JA, Carmo-Fonseca M, Lamond AI (1994) Differential interaction of splicing snRNPs with coiled bodies and interchro-matin granules during mitosis and assembly of daughter cell nuclei. J Cell Biol 126: 11–23
Frey MR, Matera G (1995) Coiled bodies contain U7 small nuclear RNA and associate with specific DNA sequences in interphase human cells. Proc Natl Acad Sci USA 92: 5915–5919
Glyn MCP, Leitch AR (1995) The distribution of a spliceosome protein in cereal (Triticeae) interphase nuclei from cells with dif-ferent metabolic activities and through the cell cycle. Plant J 8: 531–540
González-Melendi P, Testillano PS, Ahmadian P, Fadon B, Vicente O, Risueno MC (1995) In situ characterization of the late vacuo-late microspore as a convenient stage to induce embryogenesis inCapsicum. Protoplasma 187: 60–71
——— Risueño MC (1996) New in situ approaches to study the induction of pollen embryogenesis inCapsicum annuum L. Eur J Cell Biol 69: 373–386
— Beven A, Boudonck K, Abranches R, Wells B, Dolan L, Shaw P (2000) The nucleus: a highly organized but dynamic structure. J Microsc 198:199–207
Jacobs EY, Frey MR, Wu W, Ingledue TC, Gebühr TC, Gao L, Marzluff WF, Matera AG (1999) Coiled bodies preferentially associate with U4, Uli, and U12 small nuclear RNA genes in interphase HeLa cells but not with U6 and U7 genes. Mol Biol Cell 10:1653–1663
Kingham KI, Duckett JG, Glyn MCP, Leitch AR (1995) Nuclear dif-ferentiation in the filamentous caulonema of the mossFunaria hygrometrica. New Phytol 131: 543–556
Lamond AI, Carmo-Fonseca M (1993) The coiled body. Trends Cell Biol 3: 198–204
— (1998) Structure and function in the nucleus. Science 280: 547–553
Leitch AR (2000) Higher levels of organization in the interphase nucleus of cycling and differentiated cells. Microbiol Mol Biol Rev 64:138–152
Lichter R (1982) Induction of haploid plants from isolated pollen ofBrassica napus. Z Pflanzenphysiol 105: 427–434
Lischwe MA, Ochs RL, Reddy R, Cook RG, Yeoman LC, Tan EM, Reichlin M, Busch H (1985) Purification and partial characterization of a nucleolar scleroderma antigen (Mr = 34,000; pi 8.5) rich in NG,NG-dimethylarginine. J Biol Chem 260: 14304–14310
Mascarenhas JP (1975) The biochemistry of angiosperm pollen development. Bot Rev 41: 259–314
Matera AG (1999) Nuclear bodies: multifaceted subdomains of the interchromatin space. Trends Cell Biol 9: 302–309
Ochs RL, Lischwe MA, Spohn WH, Busch H (1985) Fibrillarin: a new protein of the nucleolus identified by autoimmune sera. Biol Cell 54: 123–134
Ochs RL, Stein TJ, Tan EM (1994) Coiled bodies in the nucleolus of breast cancer cells. J Cell Sci 107: 385–399
Osteryoung KW, Sundberg H, Vierling E (1993) Poly(A) tail length of a heat shock protein RNA is increased by severe heat stress, but intron splicing is unaffected. Mol Gen Genet 239: 323–333
Pechan P, Keller WA (1988) Identification of potentially embryogénic microspores inBrassica napus. Physiol Plant 74: 377–384
Raska I, Andrade LE, Ochs RL, Chan EK, Chang CM, Roos G, Tan EM (1991) Imraunological and ultrastructural studies of the nuclear coiled body with autoimmune antibodies. Exp Cell Res 195:27–37
Sánchez-Pina MA, Kieft H, Schel JHN (1989) 2. J Cell Sci 93:615–622
Schul W, van Driel R, de Jong L (1998) Coiled bodies and U2 snRNA genes adjacent to coiled bodies are enriched in factors required for snRNA transcription. Mol Biol Cell 9: 1025–1036
Sleeman J, Lyon CE, Platani M, Kreivi JP, Lamond AI (1998) Dynamic interactions between splicing snRNPs, coiled bodies and nucleoli revealed using snRNP protein fusions to the green fluorescent protein. Exp Cell Res 243: 290–304
Smith KP, KC Carter, CV Johnson, JB Lawrence (1995) U2 and Ul snRNA gene loci associated with coiled bodies. J Cell Biochem 59: 473–485
Sokal RR, Rohlf FJ (1969) Biometry: the principles and practice of statistics in biological research. W H Freeman, San Francisco
Solymosy F, Pollak T (1993) Uridylate-rich small nuclear RNAs (UsnRNAs), their genes and pseudogenes, and UsnRNPs in plants -structure and function: a comparative approach. Crit Rev Plant Sci 12: 275–369
Straatman KR, Nijsse J, Kieft H, van Aelst AC, Schel JHN (2000) Nuclear pore dynamics during pollen development and andro-genesis inBrassica napus. Sex Plant Reprod 13: 43–51
Testillano PS, Sánchez-Pina MA, Olmedilla A, Fuchs JP, Risueno MC (1993) Characterization of the interchromatin region as the nuclear domain containing snRNPs in plant cells: a cytochemical and immunoelectron microscopy study. Eur J Cell Biol 61: 349–361
— González-Melendi P, Ahmadian P, Fadon B, Risueno MC (1995) The immunolocalization of nuclear antigens during the pollen developmental program and the induction of pollen embryogen-esis. Exp Cell Res 221: 41–54
Zirbel RM, Mathieu UR, Kurz A, Cremer T, Lichter P (1993) Evi-dence for a nuclear compartment of transcription and splicing located at the chromosome boundaries. Chromosome Res 1: 93–106
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Straatman, K.R., Schel, J.H.N. Distribution of splicing proteins and putative coiled bodies during pollen development and androgenesis in Brassica napus L.. Protoplasma 216, 191–200 (2001). https://doi.org/10.1007/BF02673871
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02673871