Abstract
Monoclonal antibodies A33/22 and La11G7 have been used to study the distribution of the corre-sponding antigens, PwA33 and La, on the lampbrush chromosome loops and nucleoplasmic structures of P. waltl oocytes, using immunofluorescence, confocal laser scanning microscopy and immunogold labeling. The results obtained with these antibodies have been compared with those obtained with the Sm-antigen-specific monoclonal antibody Y12. All these monoclonal antibodies (mAbs) labeled the matrices of the majority of normal loops along their whole length. Nucleoplasmic RNP granules showed a strong staining with the mAbs La11G7 and Y12 throughout their mass, but with the mAb A33/22, they showed only a weak peripheral labeling in the form of patches on their surface. This patchy labeling was confirmed by confocal laser scanning microscopy. Electron microscopy revealed that this patchy labeling might be due to a hitherto undescribed type of submicroscopic granular structure, around 100 nm in either dimension, formed by 10-nm particles. Such granules were observed either attached to the RNP granules or free in the nucleoplasm, but rarely in relation with the normal loop matrices. These 100-nm granules may have a role in the movement of proteins and snRNPs inside the oocyte nuclei for storage, recycling, and/or degradation. Our results also suggest that all the microscopically visible free RNP granules of the nucleoplasm of P. waltl oocytes correspond to B snurposomes. The granules forming the B (globular) loops showed a labeling pattern similar to that of B snurposomes; their possible relationship is discussed.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bachmann M, Pfeifer K, Schröder HC, Müller WEG (1987) The nucleocytoplasmic shuttling of the La antigen in CV-1 cells. Mol Biol Rep 12:239–240
Bachmann M, Schröder HC, Falke D, Müller WEG (1988) Alteration of the intracellular localization of the La protein compared with the localization of UsnRNPs. Cell Biol Int Rep 12:765–789
Bachmann M, Falke D, Schröder HC, Müller WEG (1989a) Intracellular distribution of the La antigen in CV-1 cells after Herpes Simplex Virus type 1 infection compared with the localization of U small nuclear ribonucleoprotein particles. J Gen Virol 70:881–891
Bachmann M, Pfeifer K, Schröder HC, Müller WEG (1989b) The La antigen shuttles between the nucleus and the cytoplasm in CV-1 cells. Mol Cell Biochem 85:103–114
Bachmann M, Pfeifer K, Schröder, Müller WEG (1990) Characterization of the autoantigen La as a nucleic acid-dependent. ATPase/dATPase with melting properties. Cell 60:85–93
Bellini M, Lacroix JC, Gall JG (1993) A putative zinc-binding protein on lampbrush chromosome loops. EMBO J 12:107–114
Bendayan M (1984) Protein-A gold electron microscopic immunocytochemistry: methods, applications and limitations. J Electron Micros Tech 1:243–254
Bernhard W (1969) A new staining procedure for electron microscopic cytology. J Ultrastruc Res 27:250–265
Bonnanfant-Jaïs ML, Penrad-Mobayed M, Angelier N (1991) Ultrastructural similarity in landmark loops of amphibian lampbrush chromosomes. Biol Cell 71:105–114
Boucher D, Loones MT, Pyne CK, Simon F, Abbadie C, Charlemagne J, Lacroix JC (1991) Immunolocalization of a nuclear protein bound to the sphere organelle during oogenesis and embryogenesis in Pleurodeles waltl. Roux's Arch Dev Biol 199:458–468
Bucci S, Ragghianti M, Nardi I, Bellini M, Mancino G, Lacroix JL (1993) Identification of an amphibian oocyte nuclear protein as a candidate for a role in embryonic DNA replication. Int J Dev Biol 37:509–517
Callan HG (1986) Lampbrush chromosomes. Springer, Berlin New York Heidelberg
Callan HG, Gall JG (1991) Association of RNA with the B-snurposomes and C-snurposomes of Xenopus oocyte nuclei. Chromosoma 101:69–82
Carmo-Fonseca M, Pfeifer K, Schröder HC, Fatima Vaz M, Eurico Fonseca J, Müller WEG, Bachmann M (1990) Identification of La ribonucleoproteins as a component of interchromatin granules. Exptl Cell Res 185:73–85
Dreyfuss G (1986) Structure and function of nuclear and cytoplasmic ribonucleoprotein particles. Ann Rev Cell Biol 2:459–498
Dreyfuss G, Swanson MS, Pinol-Roma S (1988) Heterogenous nuclear ribonucleoprotein particles and the pathway of mRNA formation. Trends Biochem Sci 13:86–91
Fakan S, Leser G, Martin T (1984) Ultrastructural distribution of nuclear ribonucleoproteins as visualized by immunocytochemistry in thin sections. J Cell Biol 98:358–363
Gall JG (1991) Spliceosomes and snurposomes. Science 252:1499–1500
Gall JG (1992) Organelle assembly and function in the amphibian germinal vesicle. Adv Developmental Biochem 1:1–29
Gall JG, Callan HG (1989) The sphere organelle contains small nuclear ribonucleoproteins. Proc Nat Acad Sci USA 86:6635–6639
Gottlieb E, Steitz JA (1989a) The RNA binding protein La influences both the accuracy and the efficiency of RNA polymerase II transcription in vivo. EMBO J 8:841–850
Gottlieb E, Steitz JA (1989b) Function of the mammalian La protein: evidence for its action in transcription termination by RNA polymerase III. EMBO J 8:851–861
Hamelin R, Chan EK, Tan EM (1986) Antibodies against small nuclear ribonucleoproteins immunoprecipitate complexes containing ts110 Moloney murine sarcoma virus genomic and messenger RNAs. Virology 152:87–99
Lacroix JC (1968a) Etude descriptive des chromosomes en écouvillon dans le genre Pleurodeles (Amphibien, Urodèle). Ann d'Embryol Morphogenèse 1:179–202
Lacroix JC (1968b) Variations expérimentales ou spontanées de la morphologie et de l'organisation des chromosomes en écouvillon dans le genre Pleurodeles (Amphibien, Urodèle). Ann d'Embryol Morphogenèse 1:205–248
Lacroix JC, Azzouz R, Boucher D, Abbadie C, Pyne CK, Charlemagne J (1985) Monoclonal antibodies to lampbrush chromosome antigens of Pleurodeles waltlii. Chromosoma 92:69–80
Lerner EA, Lerner MR, Janeway CA, Steitz JA (1981) Monoclonal antibodies to nucleic acid-containing cellular constituents. Probes for molecular biology and autoimmune disease. Proc Natl Acad Sci USA 78:2737–2741
Leser GP, Martin TE (1987) The major protein components of hnRNP complexes. In: Thompson EB, Papaconstantinou J (eds) DNA: Protein interactions and gene regulations.) Univ Texas. Austin, TX, USA, pp 97–106
Lührmann R, Kastner B, Bach M (1990) Structure of spliceosomal snRNPs and their role in pre-mRNA splicing. Biochim Biophys Acta 1087:265–292
Malcolm DB, Sommerville J (1974) The structure of chromosome derived ribonucleoproteins in oocytes of Triturus cristatus carnifex (Laurenti). Chromosoma 48:137–158
Martin TE, Okamura CS (1981) Immunocytochemistry of nuclear hnRNP complexes. In: Busch H (ed) The cell nucleus, vol 9. Academic Press, New York, pp 119–144
N'Da E, Bonnanfant-Jaïs ML, Penrad-Mobayed M, Angelier N (1986) Size uniformity of ribonucleoprotein matrix particles in loops of Pleurodeles waltlii lampbrush chromosomes visualized by electron microscopy. J Cell Sci 81:17–27
Penrad-Mobayed M, Bonnanfant-Jaïs ML, N'Da E, Angelier N (1986) Evidence for a particular mode of transcription in globular loops of lampbrush chromosomes of the newt Pleurodeles waltlii. Chromosoma 94:319–328
Piñol-Roma S, Dreyfuss G (1992) Shuttling of pre-messenger RNA binding proteins between nucleus and cytoplasm. Nature 355:730–732
Piñol-Roma S, Swanson MS Gall JG, Dreyfuss G (1989) A novel heterogenous nuclear RNP protein with a unique distribution on nascent transcripts. J Cell Biol 109:2575–2585
Puvion E, Viron A, Assens C, Leduc EH, Jeanteur Ph (1984) Immunocytochemical identification of Nuclear structures containing snRNPs in isolated Rat liver cells. J Ultrastruct Res 87:180–189
Ryne CK, Charlemagne J, Boucher D, Lacroix JC (1988) Light and electron microscopic immunolocalization of two nuclear antigens in the liver of Pleurodeles waltl using monoclonal antibodies. Biol Cell 64:343–352
Pyne CK, Loones MT, Lacroix JC (1989) Correlation between isolated lampbrush chromosomes and nuclear structures of Pleurodeles waltl oocytes: an electron microscopic study. Chromosoma 98:181–193
Reed R, Griffith J, Maniatis T (1988) Purification and visualization of native spliceosomes. Cell 53:949–961
Roth MB, Gall JG (1987) Monoclonal antibodies that recognize transcription unit proteins on newt lampbrush chromosomes. J Cell Biol 105:1047–1054
Roth MB, Murphy C, Gall JG (1990) A monoclonal antibody that recognizes a phosphorylated epitope stains lampbrush chromosome loops and small granules in the amphibian germinal vesicle. J Cell Biol 111:2217–2224
Scott SEM, Sommerville J (1974) Location of nuclear proteins on the chromosomes of newt oocytes. Nature 250:680–682
Tuma RS, Stolk JA, Roth MB (1993) Identification and characterization of a sphere organelle protein. J Cell Biol 122:767–773
Wu CH, Gall JG (1993) U7 small nuclear RNA in C snurposomes of the Xenopus germinal vesicle. Proc Nat Acad Sci USA 90:6257–6259
Wu Z, Murphy C, Callan HG, Gall JG (1991) Small nuclear ribonucleoproteins (snRNPs) and heterogenous nuclear ribonucleoproteins (hnRNPs) in the amphibian germinal vesicle: Loops, spheres and snurposomes. J Cell Biol 113:465–483
Zahler AM, Lane WS, Stolk JA, Roth MB (1992) SR proteins: a conserved family of pre-mRNA splicing factors. Genes Dev 6:837–847
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Pyne, C.K., Simon, F., Loones, M.T. et al. Localization of antigens PwA33 and La on lampbrush chromosomes and on nucleoplasmic structures in the oocyte of the urodele Pleurodeles waltl: Light and electron microscopic immunocytochemical studies. Chromosoma 103, 475–485 (1994). https://doi.org/10.1007/BF00337386
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00337386