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The Histochemical Journal

, Volume 21, Issue 9–10, pp 575–583 | Cite as

Cytophotometric analysis of reaction rates of succinate and lactate dehydrogenase activity in rat liver, heart muscle and tracheal epithelium

  • Cornelis J. F. Van Noorden
  • Ilse M. C. Vogels
Papers

Summary

Reaction rates of succinate and lactate dehydrogenase activity in cryostat sections of rat liver, tracheal epithelium and heart muscle were monitored by continuous measurement of formazan formation by cytophotometry at room temperature. Incubation media contained polyvinyl alcohol as tissue protectant and Tetranitro BT as final electron acceptor. Control media lacked either substrate or substrate and coenzyme. Controls were also performed by adding malonate (a competitive inhibitor of succinate dehydrogenase), pyruvate (a non-competitive inhibitor of lactate dehydrogenase), oxalate (a competitive inhibitor of lactate dehydrogenase) orN-ethylmaleimide (a blocker of SH groups). A specific malonate-sensitive linear test minus control response for succinate dehydrogenase activity was obtained in liver (1.6 µmol H2 cm−3min−1) and tracheal epithelium (0.8 µmol H2cm−3 min−1) but not in heart muscle. All variations in the incubation conditions tested did not result in a linear test minus control response in the latter tissue. Because the reaction was sensitive to malonate, it was concluded that the initial reaction rate was the specific rate of succinate dehydrogenase activity in heart muscle (9.1 µmol H2 cm−3 min−1). Test minus control reactions for lactate dehydrogenase activity were distinctly non-linear for all tissues tested. This appeared to be due to product inhibition by pyruvate generated during the reaction and therefore it was concluded that the appropriate control reaction was the test reaction in the presence of 20mm pyruvate. The initial rate of the test minus this control was the true rate of lactate dehydrogenase activity. The lactate dehydrogenase activity thus found in liver parenchyma was 5.0 µmol of H2 generated per cm3 liver tissue per min.

Keywords

Pyruvate Oxalate Succinate Lactate Dehydrogenase Polyvinyl Alcohol 
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Copyright information

© Chapman and Hall Ltd 1989

Authors and Affiliations

  • Cornelis J. F. Van Noorden
    • 1
  • Ilse M. C. Vogels
    • 1
  1. 1.Laboratory of Cell Biology and HistologyUniversity of Amsterdam, Academic Medical CentreAmsterdamThe Netherlands

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