Abstract
The distribution of peroxisomes (microbodies) in the rat nephron was studied cytochemically, using glutaraldehyde- or formaldehyde-fixed tissue, by means of α-hydroxy acid oxidase activity in light microscopy or oxidation of 3,3'-diaminobenzidine (DAB) at pH 9 in both light and electron microscopy.The two cytochemical methods show peroxisomes to be nearly sperical particles found only in cells of the proximal convoluted tubule. Lysosomes were identified in the same or parallel sections, with β-glycerophosphate or 5'-cytidylic acid as substrate. They are found in all cells of the nephron. These cytochemical methods visualize the two organelles for light microscopy; they also permit unequivocal differentiation of all kidney peroxisomes from lysosomes in electron micrographs. Peroxisomes are larger and more reactive in the cells of the pars descendens (P3 segment) of the proximal convolution, located in the outer medulla and medullary rays, than in the cells of the pars convoluta (P1 and P2 segments), situated in the cortex. In contrast, lysosomes are much smaller in the P3 segment and larger and more reactive in the P1 and P2 segments. In all cells of the proximal convolution, peroxisomes tend to be concentrated nearer the base of the cells than do lysosomes. Mitochondria in P3 cells also show low levels of DAB oxidation at pH 6, in contrast to those in P1 and P2 cells. The possibility is discussed that P3 cells possess an extramitochondrial means of oxidation in which peroxisome oxidases play an important role.
Full Text
The Full Text of this article is available as a PDF (1.8 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Allen J. M., Beard M. E., Kleinbergs S. The localization of alpha-hydroxy acid oxidase in renal microbodies. J Exp Zool. 1965 Dec;160(3):329–344. doi: 10.1002/jez.1401600310. [DOI] [PubMed] [Google Scholar]
- Baudhuin P., Beaufay H., De Duve C. Combined biochemical and morphological study of particulate fractions from rat liver. Analysis of preparations enriched in lysosomes or in particles containing urate oxidase, D-amino acid oxidase, and catalase. J Cell Biol. 1965 Jul;26(1):219–243. doi: 10.1083/jcb.26.1.219. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Beard M. E., Allen J. M. A study of properties of renal microbodies of the rat. J Exp Zool. 1968 Aug;168(4):477–489. doi: 10.1002/jez.1401680408. [DOI] [PubMed] [Google Scholar]
- Burg M., Grantham J., Abramow M., Orloff J. Preparation and study of fragments of single rabbit nephrons. Am J Physiol. 1966 Jun;210(6):1293–1298. doi: 10.1152/ajplegacy.1966.210.6.1293. [DOI] [PubMed] [Google Scholar]
- De Duve C., Baudhuin P. Peroxisomes (microbodies and related particles). Physiol Rev. 1966 Apr;46(2):323–357. doi: 10.1152/physrev.1966.46.2.323. [DOI] [PubMed] [Google Scholar]
- ERICSSON J. L. ABSORPTION AND DECOMPOSITION OF HOMOLOGOUS HEMOGLOBIN IN RENAL PROXIMAL TUBULAR CELLS. Acta Pathol Microbiol Scand Suppl. 1964;168:SUPPL 168–168:1+. [PubMed] [Google Scholar]
- ERICSSON J. L. TRANSPORT AND DIGESTION OF HEMOGLOBIN IN THE PROXIMAL TUBULE. I. LIGHT MICROSCOPY AND CYTOCHEMISTRY OF ACID PHOSPHATASE. Lab Invest. 1965 Jan;14:1–15. [PubMed] [Google Scholar]
- ERICSSON J. L., TRUMP B. F. ELECTRON MICROSCOPIC STUDIES OF THE EPITHELIUM OF THE PROXIMAL TUBULE OF THE RAT KIDNEY. I. THE INTRACELLULAR LOCALIZATION OF ACID PHOSPHATASE. Lab Invest. 1964 Nov;13:1427–1456. [PubMed] [Google Scholar]
- ERICSSON J. L., TRUMP B. F., WEIBEL J. ELECTRON MICROSCOPIC STUDIES OF THE PROXIMAL TUBULE OF THE RAT KIDNEY. II. CYTOSEGRESOMES AND CYTOSOMES: THEIR RELATIONSHIP TO EACH OTHER AND TO THE LYSOSOME CONCEPT. Lab Invest. 1965 Jul;14:1341–1365. [PubMed] [Google Scholar]
- Ericsson J. L., Trump B. F. Electron microscopic studies of the epithelium of the proximal tubule of the rat kidney. 3. Microbodies, multivesicular bodies, and the golgi apparatus. Lab Invest. 1966 Oct;15(10):1610–1633. [PubMed] [Google Scholar]
- Essner E. Endoplasmic reticulum and the origin of microbodies in fetal mouse liver. Lab Invest. 1967 Jul;17(1):71–87. [PubMed] [Google Scholar]
- Fahimi H. D. Cytochemical localization of peroxidase activity in rat hepatic microbodies (peroxisomes). J Histochem Cytochem. 1968 Aug;16(8):547–550. doi: 10.1177/16.8.547. [DOI] [PubMed] [Google Scholar]
- Graham R. C., Jr, Karnovsky M. J. The early stages of absorption of injected horseradish peroxidase in the proximal tubules of mouse kidney: ultrastructural cytochemistry by a new technique. J Histochem Cytochem. 1966 Apr;14(4):291–302. doi: 10.1177/14.4.291. [DOI] [PubMed] [Google Scholar]
- Griffith L. D., Bulger R. E., Trump B. F. The ultrastructure of the functioning kidney. Lab Invest. 1967 Feb;16(2):220–246. [PubMed] [Google Scholar]
- HRUBAN Z., SWIFT H. URICASE: LOCALIZATION IN HEPATIC MICROBODIES. Science. 1964 Dec 4;146(3649):1316–1318. doi: 10.1126/science.146.3649.1316. [DOI] [PubMed] [Google Scholar]
- Hruban Z., Swift H., Slesers A. Ultrastructural alterations of hepatic microbodies. Lab Invest. 1966 Dec;15(12):1884–1901. [PubMed] [Google Scholar]
- Jacobson N. O., Jorgensen F., Thomsen A. C. On the localization of some phosphatases in three different segments of the proximal tubules in the rat kidney. J Histochem Cytochem. 1967 Aug;15(8):456–469. doi: 10.1177/15.8.456. [DOI] [PubMed] [Google Scholar]
- LONGLEY J. B., FISHER E. R. Alkaline phosphatase and the periodic acid Schiff reaction in the proximal tubule of the vertebrate kidney; a study in segmental differentiation. Anat Rec. 1954 Sep;120(1):1–21. doi: 10.1002/ar.1091200102. [DOI] [PubMed] [Google Scholar]
- LUFT J. H. Improvements in epoxy resin embedding methods. J Biophys Biochem Cytol. 1961 Feb;9:409–414. doi: 10.1083/jcb.9.2.409. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Levy M. R., Hunt A. E. L-alpha-hydroxy acid oxidase activity in Tetrahymena. Change with physiological state. J Cell Biol. 1967 Sep;34(3):911–915. doi: 10.1083/jcb.34.3.911. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maunsbach A. B. Absorption of I-125-labeled homologous albumin by rat kidney proximal tubule cells. A study of microperfused single proximal tubules by electron microscopic autoradiography and histochemistry. J Ultrastruct Res. 1966 Jun;15(3):197–241. doi: 10.1016/s0022-5320(66)80108-9. [DOI] [PubMed] [Google Scholar]
- Maunsbach A. B. Observations on the segmentation of the proximal tubule in the rat kidney. Comparison of results from phase contrast, fluorescence and electron microscopy. J Ultrastruct Res. 1966 Oct;16(3):239–258. doi: 10.1016/s0022-5320(66)80060-6. [DOI] [PubMed] [Google Scholar]
- Maunsbach A. B. Observations on the ultrastructure and acid phosphatase activity of the cytoplasmic bodies in rat kidney proximal tubule cells. With a comment on their classification. J Ultrastruct Res. 1966 Oct;16(3):197–238. doi: 10.1016/s0022-5320(66)80059-x. [DOI] [PubMed] [Google Scholar]
- Maunsbach A. B. The influence of different fixatives and fixation methods on the ultrastructure of rat kidney proximal tubule cells. I. Comparison of different perfusion fixation methods and of glutaraldehyde, formaldehyde and osmium tetroxide fixatives. J Ultrastruct Res. 1966 Jun;15(3):242–282. doi: 10.1016/s0022-5320(66)80109-0. [DOI] [PubMed] [Google Scholar]
- NACHLAS M. M., TSOU K. C., DE SOUZA E., CHENG C. S., SELIGMAN A. M. Cytochemical demonstration of succinic dehydrogenase by the use of a new p-nitrophenyl substituted ditetrazole. J Histochem Cytochem. 1957 Jul;5(4):420–436. doi: 10.1177/5.4.420. [DOI] [PubMed] [Google Scholar]
- NOVIKOFF A. B., SHIN W. Y., DRUCKER J. Mitochondrial localization of oxidative enzymes: staining results with two tetrazolium salts. J Biophys Biochem Cytol. 1961 Jan;9:47–61. doi: 10.1083/jcb.9.1.47. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Neustein H. B. Hemoglobin absorption in the proximal tubules of the kidney in the rabbit. J Ultrastruct Res. 1967 Mar;17(5):565–587. doi: 10.1016/s0022-5320(67)80141-2. [DOI] [PubMed] [Google Scholar]
- Neustein H. B., Maunsbach A. B. Hemoglobin absorption by proximal tubule cells of the rabbit kidney. A study by electron microscopic autoradiography. J Ultrastruct Res. 1966 Sep;16(1):141–157. doi: 10.1016/s0022-5320(66)80028-x. [DOI] [PubMed] [Google Scholar]
- REYNOLDS E. S. The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J Cell Biol. 1963 Apr;17:208–212. doi: 10.1083/jcb.17.1.208. [DOI] [PMC free article] [PubMed] [Google Scholar]
- SABATINI D. D., BENSCH K., BARRNETT R. J. Cytochemistry and electron microscopy. The preservation of cellular ultrastructure and enzymatic activity by aldehyde fixation. J Cell Biol. 1963 Apr;17:19–58. doi: 10.1083/jcb.17.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
- STERNBERG W. H., FARBER E., DUNLAP C. E. Histochemical localization of specific oxidative enzymes. II. Localization of diphosphopyridine nucleotide and triphosphopyridine nucleotide diaphorases and the succindehydrogenase system in the kidney. J Histochem Cytochem. 1956 May;4(3):266–283. doi: 10.1177/4.3.266. [DOI] [PubMed] [Google Scholar]
- STRAUS W. CYTOCHEMICAL OBSERVATIONS ON THE RELATIONSHIP BETWEEN LYSOSOMES AND PHAGOSOMES IN KIDNEY AND LIVER BY COMBINED STAINING FOR ACID PHOSPHATASE AND INTRAVENOUSLY INJECTED HORSERADISH PEROXIDASE. J Cell Biol. 1964 Mar;20:497–507. doi: 10.1083/jcb.20.3.497. [DOI] [PMC free article] [PubMed] [Google Scholar]
- STRAUS W. Cytochemical investigation of phagosomes and related structures in cryostat sections of the kidney and liver of rats after intravenous administration of horseradish peroxidase. Exp Cell Res. 1962 Jun;27:80–94. doi: 10.1016/0014-4827(62)90045-9. [DOI] [PubMed] [Google Scholar]
- STRAUS W. OCCURRENCE OF PHAGOSOMES AND PHAGO-LYSOSOMES IN DIFFERENT SEGMENTS OF THE NEPHRON IN RELATION TO THE REABSORPTION, TRANSPORT, DIGESTION, AND EXTRUSION OF INTRAVENOUSLY INJECTED HORSERADISH PEROXIDASE. J Cell Biol. 1964 Jun;21:295–308. doi: 10.1083/jcb.21.3.295. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Seligman A. M., Karnovsky M. J., Wasserkrug H. L., Hanker J. S. Nondroplet ultrastructural demonstration of cytochrome oxidase activity with a polymerizing osmiophilic reagent, diaminobenzidine (DAB). J Cell Biol. 1968 Jul;38(1):1–14. doi: 10.1083/jcb.38.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Seligman A. M., Ueno H., Morizono Y., Wasserkrug H. L., Katzoff L., Hanker J. S. Electron microscopic demonstration of dehydrogenase activity with a new osmiophilic ditetrazolium salt (TC-NBT). J Histochem Cytochem. 1967 Jan;15(1):1–13. doi: 10.1177/15.1.1. [DOI] [PubMed] [Google Scholar]
- Shnitka T. K. Comparative ultrastructure of hepatic microbodies in some mammals and birds in relation to species differences in uricase activity. J Ultrastruct Res. 1966 Dec;16(5):598–625. doi: 10.1016/s0022-5320(66)80009-6. [DOI] [PubMed] [Google Scholar]
- Straus W. Changes in intracellular location of small phagosomes (micropinocytic vesicles) in kidney and liver cells in relation to time after injection and dose of horseradish peroxidase. J Histochem Cytochem. 1967 Jul;15(7):381–393. doi: 10.1177/15.7.381. [DOI] [PubMed] [Google Scholar]
- Straus W. Methods for the study of small phagosomes and their relationship to lysosomes with horseradish peroxidase as a "marker protein". J Histochem Cytochem. 1967 Jul;15(7):375–380. doi: 10.1177/15.7.375. [DOI] [PubMed] [Google Scholar]
- Tolbert N. E., Oeser A., Kisaki T., Hageman R. H., Yamazaki R. K. Peroxisomes from spinach leaves containing enzymes related to glycolate metabolism. J Biol Chem. 1968 Oct 10;243(19):5179–5184. [PubMed] [Google Scholar]
- Tsukada H., Mochizuki Y., Fujiwara S. The nucleoids of rat liver cell microbodies. Fine structure and enzymes. J Cell Biol. 1966 Mar;28(3):449–460. doi: 10.1083/jcb.28.3.449. [DOI] [PMC free article] [PubMed] [Google Scholar]
- WACHSTEIN M. Histochemical staining reactions of the normally functioning and abnormal kidney. J Histochem Cytochem. 1955 Jul;3(4):246–270. doi: 10.1177/3.4.246. [DOI] [PubMed] [Google Scholar]
- WACHSTEIN M., MEISEL E. Histochemistry of hepatic phosphatases of a physiologic pH; with special reference to the demonstration of bile canaliculi. Am J Clin Pathol. 1957 Jan;27(1):13–23. doi: 10.1093/ajcp/27.1.13. [DOI] [PubMed] [Google Scholar]
- WALKER D. G., SELIGMAN A. M. The use of formalin fixation in the cytochemical demonstration of succinic and DPN- and TPN-dependent dehydrogenases in mitochondria. J Cell Biol. 1963 Mar;16:455–469. doi: 10.1083/jcb.16.3.455. [DOI] [PMC free article] [PubMed] [Google Scholar]
