Ultrastructural localization of peroxidase in leukocytes of rat bone marrow and blood

Abstract

The differentiation of leukocytes in the bone marrow and blood of normal adult male rats was studied by electron microscopy and peroxidase cytochemistry. Tissue samples were fixed in glutaraldehyde, or paraformaldehyde-glutaraldehyde, and incubated in a peroxidase medium containing 3,3'-diaminobenzidine and H2O2 ad pH 7.6. Mature cells of blood were identified, and then the earlier stages of maturation in bone marrow were analyzed. In immature cells of four cell lines, neutrophils, monocytes, basophils, and eosinophils, peroxidase is synthesized and could be demonstrated in the rough endoplasmic reticulum (RER), Golgi complex, and in cytoplasmic granules. Later in maturation, reaction product for peroxidase could not be found in RER or Golgi complex, indicating that peroxidase synthesis had ceased. In two cell lines, neutrophils and monocytes, peroxidase-negative granules were formed, and the mature cells contained two populations of cytochemically distinct granules. All granules of mature eosinophils were peroxidase-positive. In mature basophils, some granules were clearly peroxidase-positive; others displayed variable density, making interpretation uncertain. Mast cells were never seen in blood, but were abundant in bone marrow; peroxidase was never found in their granules by either electron microscopic cytochemistry or a variety of light microscopic methods. Hence, these cells differ from basophils, not only in morphology but also in the enzyme content of their granules.

The preosteoclast and its cytodifferentiation into the osteoclast: Ultrastructural and histochemical studies of rat fetal parietal bone.

“…In the present study, endogenous peroxidase activity was studied cytochemically in order to elucidate the relationship between monocytes and preosteoclasts or osteoclasts, as this activity was found to be located at various sites and intensities in the mononuclear phagocytic cells during their life cycles (BENTFELD et al, 1977;DAEMS et al, 1975; VAN DER RHEE et al, 1977;ROBBINS et al, 1971). From the results of this study, intensive peroxidase activity was detected only in the monocyte granules, and not in the preosteoclast or the osteoclast, except for the mitochondria, which may due to a cytochrome oxidase reaction.…”

In situ heparin‐induced peroxisomal reticulum and biogenesis of peroxisomes in pulmonary intravascular macrophages (PIMs) of caprine lung: an ultrastructural and cytochemical study

“…After staining with DAB and H 2 O 2 in the incubation medium, we noted a strong expression of catalase in the azurophilic granules of the PMN and dense bodies of the platelets (Bentfield et al, 1977) (Fig. 14).…”

“…Taken together, these images may represent de novo biogenesis of peroxisomal structures. Therefore, microperoxisomes from small vesicles to spherical, tubular, and lenticular subforms may well be a model for the sequential multistep assembly pathway (Titorenko and Rachubinski, 1998;Titorenko et al, 2000;Bentfield et al, 1977) initiated at the PR, a transiently formed reticulum (Schrader et al, 2000). units into the vacuolar system, the reconstitution of the coat began at 2 hr post-heparin treatment.…”

Identification of leukocytes in healthy rat gingivae by endogenous peroxidase.

“…On the other hand, Attstrom et al (2) found neutrophils, lymphocytes and monocytes in the crevices of healthy gingivae, but ultrastructural and cytochemical details of the leukocytes have not yet been described because they prov ded only morpho-logical evidence at the light microscopic level. Therefore, the present study using the cytochemical diaminobenzidine method (5,11), which is specific for each leukocyte, was designed to 1) examine whether a variety of leukocytes are localized in the junctional epithelium of healthy gingivae, and 2) elucidate ultrastructural and cytochemical natures of the leukocytes.…”