Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1981 Feb 1;88(2):274–280. doi: 10.1083/jcb.88.2.274

Application of scanning electron microscopy to x-ray analysis of frozen- hydrated sections. III. Elemental content of cells in the rat renal papillary tip

PMCID: PMC2111754  PMID: 7204493

Abstract

The electrolyte and water content of cellular and interstitial compartments in the renal papilla of the rat was determined by x-ray microanalysis of frozen-hydrated tissue sections. Papillae from rats on ad libitum water were rapidly frozen in a slush of Freon 12, and sectioned in a cryomicrotome at -30 to -40 degrees C. Frozen 0.5- micrometer sections were mounted on carbon-coated nylon film over a Be grid, transferred cold to the scanning microscope, and maintained at - 175 degrees C during analysis. The scanning transmission mode was used for imaging. Structural preservation was of good quality and allowed identification of tissue compartments. Tissue mass (solutes + water) was determined by continuum radiation from regions of interest. After drying in the SEM, elemental composition of morphologically defined compartments (solutes) was determined by analysis of specific x-rays, and total dry mass by continuum. Na, K, Cl, and H2O contents in collecting-duct cells (CDC), papillary epithelial cells (PEC), and interstitial cells (IC) and space were measured. Cells had lower water content (mean 58.7%) than interstitium (77.5%). Intracellular K concentrations (millimoles per kilogram wet weight) were unremarkable (79-156 mm/kg wet weight); P was markedly higher in cells than in interstitium. S was the same in all compartments. Intracellular Na levels were extremely high (CDC, 344 +/- 127 SD mm/kg wet weight; PEC, 287 +/- 105; IC, 898 +/- 194). Mean interstitial Na was 590 +/- 119 mm/kg wet weight. CI values paralleled those for Na. If this Na is unbound, then these data suggest that renal papillary interstitial cells adapt to their hyperosmotic environment by a Na-uptake process.

Full Text

The Full Text of this article is available as a PDF (786.4 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. APPELBOOM J. W., BRODSKY W. A., SCOTT W. N. EFFECT OF OSMOTIC DIURESIS ON INTRARENAL SOLUTES IN DIABETES INSIPIDUS AND HYDROPENIA. Am J Physiol. 1965 Jan;208:38–45. doi: 10.1152/ajplegacy.1965.208.1.38. [DOI] [PubMed] [Google Scholar]
  2. Atherton J. C., Evans J. A., Green R., Thomas S. Influence of variations in hydration and in solute excretion of the effects of lysine-vasopressin infusion on urinary and renal tissue composition in the conscious rat. J Physiol. 1971 Mar;213(2):311–327. doi: 10.1113/jphysiol.1971.sp009384. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Atherton J. C., Green R., Thomas S. Influence of lysine-vasopressin dosage on the time course of changes in renal tissue and urinary composition in the conscious rat. J Physiol. 1971 Mar;213(2):291–309. doi: 10.1113/jphysiol.1971.sp009383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Atherton J. C., Hai M. A., Thomas S. The time course of changes in renal tissue composition during mannitol diuresis in the rat. J Physiol. 1968 Jul;197(2):411–428. doi: 10.1113/jphysiol.1968.sp008567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Atherton J. C., Hai M. A., Thomas S. The time course of changes in renal tissue composition duruig water diuresis in the rat. J Physiol. 1968 Jul;197(2):429–443. doi: 10.1113/jphysiol.1968.sp008568. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Atherton J. C. Lability of renal papillary tissue composition in the rat. J Physiol. 1978 Jan;274:323–328. doi: 10.1113/jphysiol.1978.sp012150. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Battilana C. A., Dobyan D. C., Lacy F. B., Bhattacharya J., Johnston P. A., Jamison R. L. Effect of chronic potassium loading on potassium secretion by the pars recta or descending limb of the juxtamedullary nephron in the rat. J Clin Invest. 1978 Nov;62(5):1093–1103. doi: 10.1172/JCI109215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Bonventre J. V., Karnovsky M. J., Lechene C. P. Renal papillary epithelial morphology in antidiuresis and water diuresis. Am J Physiol. 1978 Jul;235(1):F69–F76. doi: 10.1152/ajprenal.1978.235.1.F69. [DOI] [PubMed] [Google Scholar]
  9. Bulger R. E., Nagle R. B. Ultrastructure of the interstitium in the rabbit kidney. Am J Anat. 1973 Feb;136(2):183–203. doi: 10.1002/aja.1001360206. [DOI] [PubMed] [Google Scholar]
  10. Bulger R. E., Trump B. F. Fine structure of the rat renal papilla. Am J Anat. 1966 May;118(3):685–721. doi: 10.1002/aja.1001180302. [DOI] [PubMed] [Google Scholar]
  11. CROSS R. B. THE EFFECTS OF OSMOTIC DIURESIS AND VASOPRESSIN UPON THE DISTRIBUTION OF SODIUM, POTASSIUM AND UREA IN THE RAT KIDNEY. Aust J Exp Biol Med Sci. 1964 Aug;42:523–538. doi: 10.1038/icb.1964.49. [DOI] [PubMed] [Google Scholar]
  12. Cameron I. L., Smith N. K., Pool T. B. Element concentration changes in mitotically active and postmitotic enterocytes. An x-ray microanalysis study. J Cell Biol. 1979 Feb;80(2):444–450. doi: 10.1083/jcb.80.2.444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. GOTTSCHALK C. W., MYLLE M. Micropuncture study of the mammalian urinary concentrating mechanism: evidence for the countercurrent hypothesis. Am J Physiol. 1959 Apr;196(4):927–936. doi: 10.1152/ajplegacy.1959.196.4.927. [DOI] [PubMed] [Google Scholar]
  14. Gardner K. D., Jr, Vierling J. M. Solids, water, and solutes in papillary region of the rat kidney. Am J Physiol. 1969 Jul;217(1):58–64. doi: 10.1152/ajplegacy.1969.217.1.58. [DOI] [PubMed] [Google Scholar]
  15. Hai M. A., Thomas S. The time-course of changes in renal tissue composition during lysine vasopressin infusion in the rat. Pflugers Arch. 1969;310(4):297–317. doi: 10.1007/BF00587241. [DOI] [PubMed] [Google Scholar]
  16. LEVITIN H., GOODMAN A., PIGEON G., EPSTEIN F. H. Composition of the renal medulla during water diuresis. J Clin Invest. 1962 May;41:1145–1151. doi: 10.1172/JCI104567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Moreton R. B., Echlin P., Gupta B. L., Hall T. A., Weis-Fogh T. Preparation of frozen hydrated tissue sections for x-ray microanalysis in the scanning electron microscope. Nature. 1974 Jan 11;247(5436):113–115. doi: 10.1038/247113a0. [DOI] [PubMed] [Google Scholar]
  18. SAIKIA T. C. COMPOSITION OF THE RENAL CORTEX AND MEDULLA OF RATS DURING WATER DIURESIS AND ANTIDIURESIS. Q J Exp Physiol Cogn Med Sci. 1965 Apr;50:146–157. doi: 10.1113/expphysiol.1965.sp001777. [DOI] [PubMed] [Google Scholar]
  19. Saubermann A. J., Beeuwkes R., 3rd, Peters P. D. Application of scanning electron microscopy to x-ray analysis of frozen-hydrated sections. II. Analysis of standard solutions and artificial electrolyte gradients. J Cell Biol. 1981 Feb;88(2):268–273. doi: 10.1083/jcb.88.2.268. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Saubermann A. J., Echlin P., Peters P. D., Beeuwkes R., 3rd Application of scanning electron microscopy to x-ray analysis of frozen-hydrated sections. I. Specimen handling techniques. J Cell Biol. 1981 Feb;88(2):257–267. doi: 10.1083/jcb.88.2.257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Schmidt-Nielsen B. Comparative physiology of cellular ion and volume regulation. J Exp Zool. 1975 Oct;194(1):207–219. doi: 10.1002/jez.1401940114. [DOI] [PubMed] [Google Scholar]
  22. Somlyo A. P., Somlyo A. V., Shuman H. Electron probe analysis of vascular smooth muscle. Composition of mitochondria, nuclei, and cytoplasm. J Cell Biol. 1979 May;81(2):316–335. doi: 10.1083/jcb.81.2.316. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Somlyo A. V., Shuman H., Somlyo A. P. Elemental distribution in striated muscle and the effects of hypertonicity. Electron probe analysis of cryo sections. J Cell Biol. 1977 Sep;74(3):828–857. doi: 10.1083/jcb.74.3.828. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. WIRZ H., HARGITAY B., KUHN W. Lokalisation des Konzentrierungsprozesses in der Niere durch direkte Kryoskopie. Helv Physiol Pharmacol Acta. 1951 Jun;9(2):196–207. [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES