Skip to main content

Advertisement

SpringerLink
Log in

A new tool for plant cell biology: in vivo antibody uptake in plant protoplasts

  • Cell Biology and Morphogenesis
  • Published:
Plant Cell Reports Aims and scope Submit manuscript

Abstract

We report on the in vivo uptake of antibodies into plant protoplasts. When protoplasts of sunflower, Arabidopsis or tobacco were incubated in vivo with an antibody, this antibody was detected by immunofluorescence in the cytoplasm and/or the nucleus, depending on the location of the target protein. Furthermore, when protoplasts were cultured in the presence of antibodies, specific effects were observed. Incubation with antibodies raised against p34cdc2 led to a strong inhibition of the division rate, and a decrease in the average DNA content of protoplasts. With antibodies against HaWLIM1, a LIM domain protein of the CRP type, a negative effect on actin organisation was observed. We conclude that antibodies can penetrate plant protoplasts in vivo, and thus may be used as powerful tools for the study of protein function.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1 Fig. 2 Fig. 3 Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Abe Y, Nakayama K, Yamanaka A, Sakurai T, Goto K (2000) Subtype-specific trafficking of endothelin receptors. J Biol Chem 275:8664–8671

    Article  CAS  PubMed  Google Scholar 

  • Adamus G, Machnicki M, Seigel GM (1997) Apoptotic retinal cell death induced by antirecoverin autoantibodies of cancer-associated retinopathy. Invest Ophthalmol Vis Sci 38:283–291

    CAS  PubMed  Google Scholar 

  • Bach I (2000) The LIM domain: regulation by association. Mech Dev 91:5–17

    Article  CAS  PubMed  Google Scholar 

  • Barbier-Brygoo H, Ephritikhine G, Klämbt D, Ghislain M, Guern J (1989) Functional evidence for an auxin receptor at the plasmalemma of tobacco mesophyll protoplasts. Proc Natl Acad Sci USA 86:891–895

    CAS  Google Scholar 

  • Barbier-Brygoo H, Ephritikhine G, Klämbt D, Maurel C, Palme K, Schell J, Guern J (1991) Perception of the auxin signal at the plasma membrane of tobacco mesophyll protoplasts. Plant J 1:83–93

    CAS  Google Scholar 

  • Baron-Epel O, Charyal PK, Schindler M (1988) Pectins as mediators of wall porosity in soybean cells. Planta 175:389–395

    CAS  Google Scholar 

  • Batelli MG, Buonamici L, Bolognesi A, Stirpe F (1994) In vivo and in vitro uptake of an anti-CD30/saporin immunotoxin by rat liver parenchymal and nonparenchymal cells. Hepatology 20:940–947

    PubMed  Google Scholar 

  • Braun M (2001) Association of spectrin-like proteins with the actin-organized aggregate of endoplasmic reticulum in the Spitzenkorper of gravitropically tip-growing plant cells. Plant Physiol 125:1611–1619

    Article  CAS  PubMed  Google Scholar 

  • Brière C, Bordel AC, Barthou H, Jauneau A, Steinmetz A, Alibert G, Petitprez M (2003) Is the LIM-domain protein HaWLIM1 associated with cortical microtubules in sunflower protoplasts? Plant Cell Physiol 44:1055–1063

    Article  PubMed  Google Scholar 

  • Chanabé C, Burrus M, Alibert G (1989) Factors affecting the improvement of colony formation from sunflower protoplasts. Plant Sci 64:125–132

    Article  Google Scholar 

  • Chanabé C, Burrus M, Bidney D, Alibert G (1991) Studies on plant regeneration from protoplasts in the genes Helianthus. Plant Cell Rep 9:635–638

    Google Scholar 

  • Colasanti J, Cho SO, Wick S, Sundaresan V (1993) Localisation of the functional p34cdc2 homolog of maize in root tip and stomatal complex cells: association with predicted division sites. Plant Cell 5:1101–1111

    Article  CAS  PubMed  Google Scholar 

  • Doedens JR, Black RA (2000) Stimulation-induced down-regulation of tumor necrosis factor-alpha converting enzyme. J Biol Chem 275:14598–14607

    Article  CAS  PubMed  Google Scholar 

  • Draetta G, Beach D (1988) Activation of cdc2 protein kinase during mitosis in human cells: cell cycle dependent phosporylation and subunit rearrangement. Cell 54:17–26

    CAS  PubMed  Google Scholar 

  • Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50:151–158

    CAS  PubMed  Google Scholar 

  • Gehring CA, McConchie RM, Venis MA, Parish RW (1998) Auxin-binding-protein antibodies and peptides influence stomatal opening and alter cytoplasmic pH. Planta 205:581–586

    Article  CAS  PubMed  Google Scholar 

  • Halin C, Niesner U, Villani ME, Zardi L, Neri D (2002) Tumor-targeting properties of antibody-vascular endothelial growth factor fusion proteins. Int J Cancer 102:109–116

    Article  CAS  PubMed  Google Scholar 

  • Hemerly A, Engler Jde A, Bergounioux C, van Montagu M, Engler G, Inze D, Ferreira P (1995) Dominant negative mutants of the Cdc2 kinase uncouple cell division from iterative plant development. EMBO J 14:3925–3936

    CAS  PubMed  Google Scholar 

  • Hjelstuen MH, Rasch-Halvorsen K, Bruland OS, De L Davies C (1998) Uptake, penetration, and binding of monoclonal antibodies with increasing affinity in human osteosarcoma multicell spheroids. Anticancer Res 18:3153–3161

    CAS  PubMed  Google Scholar 

  • Kairemo KJA, Karnani P, Jekunen AP (1996) Modulation of antibody kinetics by the cell membrane active agent Tween 80 in vivo. Anticancer Res 16:3542–3550

    CAS  PubMed  Google Scholar 

  • Kairemo KJA, Lappalainen AK, Kaapa E, Laitinen OM, Hyytinen T, Karonen SL, Gronblad M (2001) In vivo detection of intervertebral disk injury using a radiolabeled monoclonal antibody against keratan sulfate. J Nucl Med 42:476–482

    CAS  PubMed  Google Scholar 

  • Kubitscheck U, Homann U, Thiel G (2000) Osmotically evoked shrinking of guard-cell protoplasts causes vesicular retrieval of plasma membrane into the cytoplasm. Planta 210:423–431

    CAS  PubMed  Google Scholar 

  • Lai JH, Ho LJ, Kwan CY, Chang DM, Lee TC (1999) Plant alkaloid tetrandrine and its analog block CD28-costimulated activities of human peripheral blood T cells: potential immunosuppressants in transplantation immunology. Transplantation 68:1383–1392

    Article  CAS  PubMed  Google Scholar 

  • Leonhardt N, Bazin I, Richaud P, Marin E, Vavasseur A, Forestier C (2001) Antibodies to the CFTR modulate the turgor pressure of guard cell protoplasts via slow anion channels. FEBS Lett 494:15–18

    Article  CAS  PubMed  Google Scholar 

  • Lewin B (1990) Driving the cell cycle: M phase kinase, its partners, and substrates. Cell 61:743–752

    CAS  PubMed  Google Scholar 

  • Linsmaier EM, Skoog F (1965) Organic growth factor requirements of tobacco tissue cultures. Physiol Plant 18:100–127

    CAS  Google Scholar 

  • Michalak M, Milner RE, Burns K, Opas M (1992) Calreticulin. Biochem J 285:681–692

    CAS  PubMed  Google Scholar 

  • Mironov V, De Veylder L, Van Montagu M, Inze D (1999) Cyclin-dependent kinases and cell division in plants—the nexus. Plant Cell Rep 11:509–521

    Article  CAS  Google Scholar 

  • Moreau S, Thomson RM, Kaiser N, Trevaski B, Guerinot ML, Udvardi MK, Puppo A, Day DA (2002) GmZip encodes a symbiosis-specific zinc transporter in soybean. J Biol Chem 277:4738–4746

    Article  CAS  PubMed  Google Scholar 

  • Mundel C, Baltz R, Eliasson A, Bronner R, Grass N, Krauter R, Evrard JL, Steinmetz A (2000) A LIM-domain protein from sunflower is localized to the cytoplasm and/or nucleus in a wide variety of tissues and is associated with the phragmoplast in dividing cells. Plant Mol Biol 42:291–302

    CAS  PubMed  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Plant 15:473–497

    CAS  Google Scholar 

  • Neuhaus J-M, Boevink P (2001) The fluorescent protein (GFP) as reporter in plant cells. In: Hawes C, Satiat-Jeunemaitre B (ed) Plant cell biology. Oxford University Press, Oxford, p 338

  • Reed SI (1991) G1-specific cyclins: in search of an S-phase-promoting factor. Trends Genet 7:95–99

    Google Scholar 

  • Rück A, Palme K, Venis MA, Napier RM, Felle HH (1993) Patch-clamp analysis establishes a role for an auxin binding protein in the auxin stimulation of plasma membrane current in Zea mays protoplasts. Plant J 4:41–46

    Article  Google Scholar 

  • Shutian R, O’Donoghue JA, Larson SM, Finn RD, Jungbluth A, Welt S, Humm JL (2000) Optimizing the sequence of combination therapy with radiolabeled antibodies and fractionated external beam. J Nucl Med 41:1905–1912

    CAS  PubMed  Google Scholar 

  • Walter J, Fluhrer R, Hartung B, Willem M, Kaether C, Capell A, Lammich S, Multhaup G, Haass C (2001) Phosphorylation regulates intracellular trafficking of beta-secretase. J Biol Chem 276:14634–14641

    CAS  PubMed  Google Scholar 

  • Zhu H, Baxter LT, Jain RK (1997) Potential and limitations of radioimmunodetection and radioimmunotherapy with monoclonal antibodies. J Nucl Med 38:731–741

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

The authors thank Philippe Anson and Marie-Josée Tavella for their technical assistance in plant culture and protoplast preparation.

Author information

Authors and Affiliations

  1. UMR CNRS/UPS 5546, BP 17 Auzeville, 31326, Castanet-Tolosan, France

    C. Brière

  2. Laboratoire “Biotechnologie et Amélioration des Plantes”, INP-ENSAT, BP 107 Auzeville, 31326, Castanet-Tolosan, France

    H. Barthou & M. Petitprez

Authors
  1. C. Brière
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. H. Barthou
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M. Petitprez
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to C. Brière.

Additional information

Communicated by M. Beckert

Rights and permissions

Reprints and permissions

About this article

Cite this article

Brière, C., Barthou, H. & Petitprez, M. A new tool for plant cell biology: in vivo antibody uptake in plant protoplasts. Plant Cell Rep 22, 878–884 (2004). https://doi.org/10.1007/s00299-004-0788-9

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00299-004-0788-9

Keywords

Search

Navigation