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Glucose-6-phosphate dehydrogenase activity in spinach as measured by image analysis: A new approach for plant enzyme histochemistry

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Summary

A relatively low-cost computer-assisted image analysis system is described. Software has been specifically written for the continuous monitoring of absorbance readings on cryostat sectons of plant tissues incubated in media to reveal enzyme activities. The equipment was tested by quantify ing glucose-6-phosphate dehydrogenase activity in cryostat sections from shoot apices of spinach plants. The reaction rate of the dehydrogenase activity was monitored at two incubation temperatures, 20°C and 30°C. Control incubations were performed in media lacking substrate. The specific test minus control reaction at 30°C was twice that at 20°C. Variation of the substrate concentration at 30°C yielded a Km value of 0.37 mM. These preliminary results show that our image analysis system can be used for kineti measurements of dehydrogenase activity in froizen tssue sections and constitute a new approach for enzyme histochemistry in the shoot apicla meristem.

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References

  • Albe, X., Vassilakos, P., Helfer-Guarnori, K., Givel, L. C., DeQuay, N., Suardet, L., Eliason, J. F. &Odartchenko, N. (1990) Independent prognostic value of ploidy in colorectal cancer. A prospective sudy using image cytometry.Cancer 66, 1168–75.

    CAS  PubMed  Google Scholar 

  • Altman, F. P. (1975) Quantitation in histochemistry: a review of some commercially available microdensitometers.Histochem. J. 7, 375–95.

    Article  CAS  PubMed  Google Scholar 

  • Auderset, G., Gahan, P. B., Dawson, A. L. &Greppin, H. (1980) Glucose-6-phosphate dehydrogenase as an early marker of floral induction in shoot apices ofSpinacia oleracea var. Nobel.Plant Sci. Lett. 20, 109–13.

    CAS  Google Scholar 

  • Auderset, G., Gahan, P. B., Onyia, G. O. C. &Greppin, H.. (1985) Increased pentose phosphate activity linke to floral induction in apices ofSpinacia oleracea during short days.Ann. Bot. 55, 61–4.

    CAS  Google Scholar 

  • Bernier, G., Kinet, J. M. &Sachs, R. M. (1981) The Physiology of Flowering. Boca Raton: CRC Press.

    Google Scholar 

  • Blanco, C. E., Sieck, G. C. &Edgerton, V. R. (1988) Quantitative histochemical determination of succinic dehydrogenase activity in skeletal muscle fibres.Histochem. J. 20, 230–43.

    Article  CAS  PubMed  Google Scholar 

  • Bodson, M. &Outlaw, W. H. Jr (1985) Elevation of sucrose content of the shoot apical meristem ofSinapis alba at floral evocation.Plant Physiol. 79, 420–4.

    CAS  Google Scholar 

  • Butcher, R. G. (1972) Precise cytochemical measurement of neotetrazolium formazan by scanning and integrating microdensitometry.Histochemistry 32, 171–90.

    CAS  PubMed  Google Scholar 

  • Butcher, R. G. (1978) The measurement in tissue sections of the two formazans derived from nitroblue tetrazolium in dehydrogenase reactions.Histochem. J. 10, 739–44.

    Article  CAS  PubMed  Google Scholar 

  • Butcher, R. G. &Van Noorden, C. J. F. (1985) Reaction rate studies of glucose-6-phosphate dehydrogenase activity in sections of rat liver using four tetrazolium salts.Histochem. J. 17, 993–1008.

    Article  CAS  PubMed  Google Scholar 

  • Chieco, P., Jonker, A., Melchiorri, C., Vanni, G. &Van Noorden, C. J. F. (1994) A user's guide for avoiding errors in absorbance image cytometry: a review with original experimental observations.Histochem. J. 26, 1–19.

    Article  CAS  PubMed  Google Scholar 

  • Crespi, P., Crévecoeur, M., Penel, C. &Greppin, H. (1993) In vitro modification of spinach plasmalemma thickness, Biol. Plant.35, 491–7.

    Google Scholar 

  • Crèvecoeur, M., Crespi, P., Lefort, F. &Greppin, H. (1992) Serols and plasmalemma modifications in spinach apex during transtion to flowering.J. Plant Physiol. 139, 595–9.

    Google Scholar 

  • Gahan, P. B. (1991) Quantative enzyme cytochemistry in plant biotechnology.Phytochem. Anal. 2, 97–106.

    CAS  Google Scholar 

  • Gahan, P. B., McLean, J., Kalina, M. &Sharma, W. (1967) Freezesectioning of plant tissues: the technique and its use in plant histochemistry.J. Exp. Bot. 18, 151–9.

    CAS  Google Scholar 

  • Gahan, P. B., Auderset, G. &Greppin, H. (1979) Pentose phosphate pathway activity during floral induction inSpinacia oleracea var. Nobel.Ann. Bot. 44, 121–4.

    CAS  Google Scholar 

  • Greppin, H. (1986) Peroxidase as a tool in the study of the flowering process. InMolecular and Physiological Aspects of Plant Peroxidases (edited byGreppin, H., Penel, C. &Gaspar, Th.) Genève: pp. 333–9.Université de Genève, Centre de Botanique.

    Google Scholar 

  • Greppin, H., Bonzon, M., Crespi, P.,Crèvecoeur, M., Degli Agosti, R. &Penel, C. (1990) Physiological macrofunctions and indicators of the flowering process. InIntra- and Intercellular Communications in Plants (edited byMillet, B.., &Greppin, H.) pp. 107–23 Paris: INRA.

    Google Scholar 

  • Hildebrand, R. &Schleicheri, A. (1986) Image analysis of the histochemical demonstration of glucose-6-phosphatase activity in rat liver.Histochemistry 86, 181–90.

    Article  CAS  PubMed  Google Scholar 

  • Jacqmard, A. (1978) Histochemical localization of enzyme acitivy during floral evocation in the shoot apical meristem ofSinapis alba.Protoplam 94, 315–24.

    CAS  Google Scholar 

  • Jonges, G. N. &Van Noorden, C. J. F. (1989)In situ kinetic parameters of glucose-6-phosphate dehydrogenase and phosphogluconate dehydrogenase in different areas of the rat liver acinus.Histochem. J. 21, 585–94.

    Article  CAS  PubMed  Google Scholar 

  • Kugler, P. (1981) Kinetic and morphometric measurements of enzyme reactions in tissue sections with a new instrumental setup.Histochemistry 71, 433–49.

    Article  CAS  PubMed  Google Scholar 

  • Labordf, K., Bussieres, L., De Smet, A., Dechaux, M. &Sachs, C. (1990) Quantification of renal Na−K-ATPase activity by image analysing system.Cytometry 11, 859–68.

    Google Scholar 

  • Lawrence, G. M., Beesley, A. H. &Matthews, J. B. (1989) The use of continous monitoring and computer-assisted image analysis for the histochemical quantification of hexokinase activity.Histochem. J. 21, 557–64.

    Article  CAS  PubMed  Google Scholar 

  • Lowry, O. H. &Passonneal, J. V. (1972) A Flexible System of Enzymatic Analysis. New York: Acadmic Press.

    Google Scholar 

  • Markovic, N., Markovic, O., Rorets, J. &Markovic S. (1994)A new assay for intracellular measurement of inosine monophosphate dehydrogenase activity: a guide for better selection of patients for enzyme-targeted chemotherapy.H. Histochem. Cytochem. 42, 23–35.

    CAS  Google Scholar 

  • Medrord, J. I. (1992) Vegatative apical meristems.Plant Cell 4 1029–39

    Google Scholar 

  • Negi, D. S. &Stephens, R. J. (1977) An improved method for the histochemical localization of glucose-6-phosphate dehydrogenase in animal and plant tissues.J. Histochem. Cytochem. 25, 149–54.

    CAS  PubMed  Google Scholar 

  • Nolte, J. &Pette, D. (1972) Microphotometric determination of enzyme activity in single cells in cryostat sections. I. Application of the gel film technique to microphotometry and studies of the intracellular distribution of succinate dehydrogenase and lactate dehydrogenase activities in rat liver.J. Histochem. Cytochem. 20, 567–76.

    CAS  PubMed  Google Scholar 

  • Nougarède, A. (1967) Experimental cytology of the shoot apical cells during vegetative growth and flowering.Int. Rev. Cytol. 21, 203–51.

    PubMed  Google Scholar 

  • Orr, A. R. (1984) Histochemical study of enzyme acitivty in the shoot apical meristem ofBrassica mapestris, L. during transition to flowering II cytochrome oxidase.Bot. Gaz. 145, 308–11.

    Article  CAS  Google Scholar 

  • Orr, A. R. (1985) Histochemical study of enzyme activity in the shoot apical meristem ofBrassica campestris L. during transition to flowering. III. Glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenas.Bot. Gaz. 146, 477–82.

    Article  CAS  Google Scholar 

  • Orr, A. R. (1987) Changes in glyceraldehyde-3-phsphate dehydrogenase activity in shoot apical metritems ofBrassica campestris during transition of flowering.Am. J. Bot. 74, 1161–6.

    CAS  Google Scholar 

  • Oultlaw, W. H. Jr (1980) Sedescriptive evaluation of quantitative histochemical methods based on pyridine nucleotides.Ann. Rev. Plant Physiol. 31, 299–311.

    Google Scholar 

  • Petersen, K. &Orr, A. (1983) Histochemical study of enzyme activity in the schoot apicla metristem ofBrassica campestris L. during transition to flowering. I. Succinic dehydrogenase.Bot. Gaz. 144, 338–41.

    CAS  Google Scholar 

  • Schlattner, U., Wagner, E., Greppin, H. &Bonzon, M. (1994) Adenylate kinase in tobacco cell cultures. II. Variability and regulation of isoform activity patterms in different cell lines.J. Plant Physiol. 144, 400–9.

    CAS  Google Scholar 

  • Sinowatz, F., Scheubeck, M., Wroibel, K. H. &Zwack, M. (1983) Histochemical localization and quantification of glucose-6-phosphate dehydrogenase in bovine Leyding cells.Histochem. J. 15, 831–44.

    Article  CAS  PubMed  Google Scholar 

  • Stoward, P. J., Altman, F. P. &Seldler, E. (1991) Principles of oxidoreductase histochemistry. InHistochemistry Theoretical and Applied (edited byStoward, P. J., Pearse, A. G. E.), Vol. III, pp. 1–25. Edinburg: Churchill Livingstone.

    Google Scholar 

  • Van Noorden, C. J. F. (1989) Principles of cytophotometry in enzyme histochemistry and validity of the reactions.Acta Histochem., Suppl.37, 21–35.

    Google Scholar 

  • Van Noorden, C. J. F. &Butcher, R. G (1991) Quantitative enzyme hitochemistry InHistochemistry, Theoretical and Applied (edited byStoward, P. J. &Pearse, A. G. E.), Vol. III, pp. 355–432, Edinburgh: Churchill Livingtone.

    Google Scholar 

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Authors and Affiliations

  1. Laboratory of Pllant Biochemistry and Physiology, University of Geneva, 3, pl de l'Université, 1211, Geneva 4, Switzerland

    Michèle Crèvecoeur, M'Baye Cissé &  Hubert Greppin

  2. Department of Pathology, 1 rue Michel Servet, 206, Geneva, Switzerland

    Xavier Albe

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  1. Michèle Crèvecoeur
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  2. M'Baye Cissé
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  3. Xavier Albe
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  4. Hubert Greppin
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Crèvecoeur, M., Cissé, M., Xavier Albe et al. Glucose-6-phosphate dehydrogenase activity in spinach as measured by image analysis: A new approach for plant enzyme histochemistry. Histochem J 28, 25–32 (1996). https://doi.org/10.1007/BF02331424

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  • DOI: https://doi.org/10.1007/BF02331424

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