Histochemistry of Lactic Dehydrogenase in Heart and Pectoralis Muscles of Rat

Baba, Nobuhisa; Sharma, Hari

doi:10.1083/jcb.51.3.621

Cited by 92 publications

(61 citation statements)

References 33 publications

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“…It is also known that purified LDH binds to intracellular proteins such as tubulin (K d ϭ 1.0 M), microtubules (K d ϭ 3.5 M) (Walsh et al 1989), and troponin (K d ϭ 0.73-2.3 M; Yasykova et al 1990). Skeletal muscle LDH has been observed to be localized in sarcoplasmic reticulum (e.g., Baba and Sharma 1971) and mitochondria (Baba and Sharma 1971;Lluis 1984;Szcz sna-Kaczmarek 1990). Mitochondria isolated from liver also contain LDH (e.g., Brandt et al 1987).…”

Section: Discussionmentioning

confidence: 99%

Effects of Tissue Protectants on the Kinetics of Lactate Dehydrogenase in Cells

Nakae

Stoward

1997

J Histochem Cytochem.

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SUMMARYWe studied the effects of two tissue protectants, polyvinyl alcohol (PVA) and agarose gel, on a kinetic parameter of lactate dehydrogenase LDH that is assumed to be related to the extent of diffusion of the enzyme out of tissue sections during its histochemical assay. The kinetics of the enzyme in mouse gastrocnemius (skeletal) muscle fibers and periportal hepatocytes were determined in unfixed sections incubated either on substrate ( L -lactate)-containing agarose gel films or in aqueous assay media in the presence or absence of 18% PVA. The absorbances of the formazan final reaction products at their isobestic point were measured continuously in the cytoplasm of individual cells as a function of incubation time, using a real-time image analysis system. Whichever incubation medium was used, the absorbances in the two cell types increased nonlinearly during the first minute of incubation but linearly for incubation times between 1 and 3 min. The nonlinearity of the LDH reaction was analyzed using the equation v i Ϫ v ϭ a Њ A , where v i is the observed initial velocity determined from the absorbance changes during the first 10 sec of incubation and v and Њ A are respectively the gradient and intercept on the absorbance axis of the linear regression line of the absorbance on incubation times between 1 and 3 min. The plots of the observed ( v i Ϫ v ) against Њ A were linear. Their gradients a were characteristic for each cell type and tissue protectant. The a values for skeletal muscle fibers were 12-43% lower than those for hepatocytes. The a value for hepatocytes obtained with the PVA method was 32% lower than that determined with the gel film method. For skeletal muscle fibers, the a values determined by the two methods were almost the same. Addition of excess pyruvate to the aqueous assay medium had no effects on a for either muscle fibers or hepatocytes. In contrast, a was zero for sections of polyacrylamide gels containing purified enzyme, whether incubated on agarose films or in PVA media. These data confirmed that the constant a is related to the extent to which the enzyme diffuses out of sections during incubation but not to product inhibition of LDH by pyruvate. PVA was more effective for protecting diffusion of LDH from hepatocytes than from skeletal muscle fibers, possibly because hepatocytes contain a greater proportion of diffusable (unbound) LDH than skeletal muscle fibers.

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Section: Discussionmentioning

confidence: 99%

Effects of Tissue Protectants on the Kinetics of Lactate Dehydrogenase in Cells

Nakae

Stoward

1997

J Histochem Cytochem.

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“…Histochemical (24,25) and immunolabeling (19) techniques have been used to verify the presence of LDH in the mitochondria. Using stable carbon isostope-labeled lactate (i.e.…”

Section: Discussionmentioning

confidence: 99%

Physical and Functional Association of Lactate Dehydrogenase (LDH) with Skeletal Muscle Mitochondria

Elustondo

White²,

Hughes³

et al. 2013

Journal of Biological Chemistry

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Background:The notion of mitochondrial lactate oxidation in skeletal muscle is controversial. Results: Mitochondrial respiration increased in the presence of substrates and cofactors for the lactate dehydrogenase (LDH) reaction. Respiration was inhibited when mitochondrial pyruvate transport was blocked. Conclusion: Extra-matrix LDH is associated with muscle mitochondria. Significance: LDH is strategically positioned within skeletal muscle fibers to functionally interact with mitochondria.

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“…The immunohistochemical demonstration of G6PD also reports a basically similar localization on the r-ER (Tanahashi and Hori, 1980). In addition, other soluble enzymes are also known positive in the cytoplasm immediately adjacent to the r-ER ultracytochemically (Watabiki et al , 1979, Fahimi and Karnovsky, 1966, Chida et al, 1975, Baba and Sharma, 1971Yamashita et al, 1979). Applying as low a concentration of glutaraldehyde as 0.25% with 1% paraformaldehyde, it might not be easy to detect G6PD as being in close relationship with the ribosomes, but just remain to say in hyaloplasm.…”

Section: Discussionmentioning

confidence: 99%