Original Article: Electron Microscopic Radioautographic Studies on the RNA Synthesis in the Colonic Epithelial Cells of Aging Mice
DOI:
https://doi.org/10.12970/2308-8044.2013.01.02.4Abstract
For the purpose of studying the aging changes of macromolecular synthesis in the colonic cells of experimental animals, we studied 10 groups of aging mice during aging from fetal day 19 to postnatal month 24. They were injected with 3H-uridine, a precursor for RNA synthesis, sacrificed and the colonic tissues were taken out, fixed and processed for light and electron microscopic radioautography. On many radioautograms the localization of silver grains demonstrating RNA synthesis in colonic epithelial cells in respective aging groups were analyzed qualitatively. The number of mitochondria per cell, the number of labeled mitochondria with silver grains in each cell in respective aging groups were analyzed qualitatively and quantitatively in relation to the aging of animals.
The results revealed that the RNA synthesis as expressed by the number of silver grains in cell nuclei, cell organelles, changed with the aging of animals. It was demonstrated that the number of mitochondria increased from embryonic day 19 to postnatal newborn day 1, 3, 7, 14, adult month 1 and 2, reaching the maximum, then decreased to senile year 1 to 2. On the other hand, the number of labeled mitochondria showing RNA synthesis at various ages increased from embryonic day 19 to postnatal newborn day 1, 3, 7, 14, adult month 1 and 2, reaching the maximum and decreased slightly to senile month 6 to senile year 1 and 2. To the contrary, the labeling index increased from embryonic day 19 to postnatal day 1, 2 and 3, then decreased to day 14 and month 1, and increased again to month 6, 12 and month 24, indicating the aging changes. These results demonstrated that intramitochondrial RNA synthesis in the colonic epithelial cells increased, then decreased and again increased due to aging of individual animals depending upon the cellular activities at respective aging stages. Based upon our findings, available literatures on macromolecular synthesis in mitochondria of various cells are reviewed.
References
Nagata T, Usuda N. Image processing of electron microscopic radioaautograms in clinical electron microscopy. J Clin Electron Microsc 1985; 18: 5-6.
Nagata T, Usuda N, Ma H. Electron microscopic radioautography of nucleic acid synthesis in pancreatic acinar cells of prenatal and postnatal aging mice. Proc. 11th Internat. Cong. Electron Microsc, Kyoto, Japan, 1986; Vol. 3: pp. 2281-2282.
Nagata T, Usuda N, Ma H. 1986; Application of high voltage electron microscopy to histochemistry of whole mount preparations of cultured cells or thick sections from embedded tissues. Proc. 11th Internat. Cong. Electron Micrsco. Kyoto, Japan, pp.1183-1184.
Nagata T, Usuda N, Ma H. Electron microscopic radioautograpahy of nucleic acid synthesis in pancreatic acinar cells of prenatal and postnatal aging mice. Proc. 11th Internat. Cong. Electron Micrsco. Kyoto, Japan, 1986; pp. 2281-2282.
Nagata T. Radiolabeling of soluble and insoluble compounds as demonstrated by light and electron microscopy. In, Recent Advances in Cellular and Molecular Biology. Vol. 6 Molecular Biology of Pyrimidines, DNA, peroxisomes, organelles and cell movements. Wegmann, R. J. and Wegmann, M. A. eds, Peeters Press, Leuven, Belgium 1992; pp. 9-21.
Nagata T. Quantitative analysis of histochemical reactions: image analysis of light and electron microscopic radioautograms. Acta Histochem Cytochem 1993; 26: 281-91. http://dx.doi.org/10.1267/ahc.26.281
Nagata T. Quantitative light and electron microscopic radioautographic studies on macromolecular synthesis in several organs of prenatal and postnatal aging mice. Chinese J Histochem Cytochem 1993; 2: 106-108.
Nagata T. Radioautographic study on intramitochondiral nucleic acid synthesis: Its relationship to the cell cycle in cultivated cells. Proc. 4th Internat. Cong. Histochem. Cytochem. Kyoto, Japan 1972; 1: 223-224.
Nagata T. Electron microscopic radioautography of intramitochondrial nucleic acid synthesis in mammalian cells in vitro. Proc. 8th Internat. Cong. Electron Microsc, Canberra, Australia 1974; 2: 346-47.
Nagata T. Electron microscopic radioautography of RNA synthesis of HeLa cells in culture. Histochemie 1972; 32: 163-70. http://dx.doi.org/10.1007/BF00303731
Nagata T, Iwadare N, Murata F. Mitochondiral and nucleolar RNA synthesis as revealed by electron microscopic radioatography. Proc. 5th Internat. Cong. Histochem. Cytochem. Bucharest, Romania 1976; 1: 242-243.
Nagata T. Radioautographology, General and Special. In, Prog. Histochem. Cytochem, Graumann, W. ed, Urban-Fischer, Jena, Germany 2002; 32(2): pp. 57-228.
Nagata T. Macromolecular synthesis in the livers of aging mice as revealed by electron microscopic radioautography. In, Prog. Histochem. Cytochem, Sasse, D, Schumacher, U, eds, Eslevier, Jena, Germany 2010; 45(1): pp. 1-80.
Nagata T. Macromolecular synthesis in mitochondria in adrenal glands of mice as observed by electron microscopy. In, DNA Microarrays, Synthesis and Synthetic DNA, Nova Science Publishers, NY, USA 2011; in press.
Nagata T, Shibata O, Nawa T. Incorporation of tritiated thymidine into mitochondrial DNA of the liver and kidney cells of chickens and mice in tissue culture. Histochemie 1967; 10: 305-308. http://dx.doi.org/10.1007/BF00304312
Nagata T. Aging changes of macromolecular synthesis in various organ systems as observed by microscopic radioautogtraphy. Methods Find Exp Clin Pharmacol 1999; 21: 683-706. http://dx.doi.org/10.1358/mf.1999.21.10.795781
Nagata T. Chapter 2, Special cytochemistry in cell biology. In, Jeon, K. W. ed, International Review of Cytology, Vol. 211, Academic Press, N. Y, USA 2001; pp. 33-154.
Nagata T. Light and electron microscopic radioautographic studies on macromolecular synthesis in amitotic hepatocytes in aging mice. Cell Mol Biol 2003; 49: 591-11.
Nagata T. Macromolecular synthesis in hepatocyte mitochondria of aging mice as revealed by electron microscopic radioautography. I: Nucleic acid synthesis. In, Modern Research and Educational Topics in Microscopy. Mendez-Vilas, A, Diaz, J. eds, Formatex Micrscopy Series No. 3, Formatex, Badajoz, Spain 2007; Vol. 1: pp. 245-258.
Nagata T. Macromolecular synthesis in hepatocyte mitochondria of aging mice as revealed by electron microscopic radioautography. II: Protein synthesis. In, Modern Research and Educational Topics in Microscopy. Mendez-Vilas, A. and Diaz, J. eds, Formatex Micrscopy Series No. 3, Formatex, Badajoz, Spain 2007; Vol. 1: pp. 259-271.
Nagata T. Sexual difference between the macromolecular synthesis of hepatocyte mitochondria in male and female mice in aging as revealed by electron microscopic radioautography. Chapter 22. In, Women and Aging: New Research, H. T. Bennninghouse, A. D. Rosset, eds. Nova Biomed. Books, New York, USA 2009; pp. 461-487.
Nagata T. Protein synthesis in hepatocytes of mice as revealed by electron microscopic radioautography. In, Protein Biosynthesis. Esterhouse, T. E. and Petrinos, L. B, eds, Nova Biomed. Books, New York, USA 2009; pp.133-161.
Nagata T. Electron microscopic radioautographic study on mitochondrial DNA synthesis in adrenal cortical cells of developing mice. J Cell Tis Res 2008; 8: 1303-12.
Nagata T. Electron microscopic radioautographic study on mitochondrial DNA synthesis in adrenal cortical cells of developing and aging mice. Sci World J 2008; 8: 683-97. http://dx.doi.org/10.1100/tsw.2008.93
Nagata T. Electron microscopic radioautographic study on mitochondrial DNA synthesis in adreno-cortical cells of aging ddY mice. Bull Shinshu Inst Alternat Med Welfare 2009; 4: 51-66.
Nagata T. Electron microscopic radioautographic study on mitochondrial RNA synthesis in adrenocortical cells of aging mice. Open Anat J 2010; 2: 91-97.
Nagata T. Electron microscopic radioautographic study on mitochondrial DNA synthesis in adrenal medullary cells of developing and aging mice. J Cell Tissue Res 2009; 9: 1793-802.
Nagata T. Electron microscopic radioautographic study on DNA synthesis of mitochondria in adrenal medullary cells of aging mice. Open Anat J 2009; 1: 14-24.
Nass S, Nass MMK. Intramitochondrial fibers with DNA characteristics. I. Fixation and electron staining reactions. II. Enzymatic and other hydrolytic treatments. J Cell Biol 1963; 19: 593-29. http://dx.doi.org/10.1083/jcb.19.3.593
Gibor A, Granick S. Plastids and mitochondria: Inheritable system. Science 1964; 145: 890-97. http://dx.doi.org/10.1126/science.145.3635.890
Gahan PB, Chayen J. Cytoplasmic deoxyribonucleic acid. Internat Rev Cytol 1965; 18: 223-47. http://dx.doi.org/10.1016/S0074-7696(08)60555-0
Nass MMK. The circularity of mitochondrial DNA. Proc Nat Acad Sci USA 1966; 56: 1215-22. http://dx.doi.org/10.1073/pnas.56.4.1215
van Bruggen EFJ, Borst P, Ruttenberg GJCM, Gruber M, Kroon AM. Circular mitochondrial DNA. Biochim Biophys Acta 1966; 119: 437-39. http://dx.doi.org/10.1016/0005-2787(66)90210-3
Sinclair JH, Stevens BJ. Circular DNA filaments from mouse mitochondria. Proc Natl Acad Sci USA 1966; 56: 508-14. http://dx.doi.org/10.1073/pnas.56.2.508
Schatz G. Biogenesis of mitochondria. In, Membranes of Mitochondria and Chloroplasts. Racker E, ed, Van Nostrand-Reinhold, New York, USA 1970; pp. 251-314.
Guttes E, Guttes S. Thymidine incorporation by mitochondria in Physarum polycephalum. Science 1964; 145: 1057-58. http://dx.doi.org/10.1126/science.145.3636.1057
Schuster FL. A deoxyribose nucleic acid component in mitochondria of Didymium nigirpes, a slime mold. Exp Cell Res 1965; 39: 329-45. http://dx.doi.org/10.1016/0014-4827(65)90038-8
Stone GE, Miller OL Jr. A stable mitochondrial DNA in Tetrahymena puriformis. Exp Zool 1965; 159: 33-37. http://dx.doi.org/10.1002/jez.1401590104
Chévremont M. Cytoplasmic deoxyribonucleic acids: Their mitochondrial localization and synthesis in somatic cells under experimental conditions and during the normal cell cycle in relation to the preparation for mitosis. Cell Growth and Cell Division. Symposia of the Internat. Soc. for Cell Biol. Vol. 2, Harris R. J. C, ed, Academic Press, New York, USA 1963; p. 323-333.
Salpeter MM, Bachmann L, Salpeter EE. Resolution in electron microscope radioautography. J Cell Biol 1969; 44: 1-20. http://dx.doi.org/10.1083/jcb.41.1.1
Nadler NJ. The interpretation of grain counts in electron microscope radioautography. J Cell Biol 1971; 49: 377-82.
Nagata T. Techniques and application of electron microscopic radioautography. J Electron Microsc 1996; 45: 258-74. http://dx.doi.org/10.1093/oxfordjournals.jmicro.a023442
Nagata T. Techniques and application of microscopic radioautography. Histol Histopathol 1997; 12: 1091-24.
Uchida K, Mizuhira V. Electron microscope autoradiography with special reference to the problem of resolution. Arch Histol Jap 1971; 31: 291-20. http://dx.doi.org/10.1679/aohc1950.31.291
Murata F, Yoshida K, Ohno S, Nagata T. Electron microscopic radioautography using a combination of phenidon developer and domestic emulsion. Acta Histochem Cytochem 1979; 12: 443-50. http://dx.doi.org/10.1267/ahc.12.443
Nagata T. Electron microscopic observation of target cells previously observed by phase-contrast microscopy: Electron microscopic radioautography of laser beam irradiated cultured cells. J Clin Electron Microsc 1984; 17: 589-90.
Nagata T, Murata F, Yoshida K, Ohno S, Iwadare N. Whole mount radioautography of cultured cells as observed by high voltage electron microscopy. Proc. 5th Internat. Conf. High Voltage Electron Microsc. Kyoto, Japan 1977b; pp. 347-350.
Nagata T, Iwadare N, Murata F. Electron microscopic radioautography of nucleic acid synthesis in cultured cells treated with several carcinogens. Acta Pharmacol Toxicol 1977; 41: 64-65.
Nagata T, Murata F. Electron microscopic dry-mounting radioautography for diffusible compounds by means of ultracryotomy. Histochemistry 1977; 54: 75-82. http://dx.doi.org/10.1007/BF00493331
Nagata T, Ohno S, Murata F. Electron microscopic dry-mounting radioautography for soluble compounds. Acta Pharmacol Toxicol 1977; 41: 62-63.
Nagata T, Nawa T, Yokota S. A new technique for electron microscopic radioautography of soluble compouds. Histochemie 1969; 18: 211-49.
Nagata T, Ito M, Chen S. Aging changes of DNA synthesis in the submandibular glands of mice as observed by light and electron microscopic radioautography. Ann Microsc 2000; 1: 13-22.
Nagata T, Ohno S, Kawahara I, Yamabayashi S, Fujii Y, Murata F. Light and electron microscopic radioautography of nucleic acid synthesis in mitochondria and peroxisomes of rat hepatic cells during and after DEHP administration. Acta Histochem Cytochem 1979; 16: 610-11.
Nagata T, Ohno S, Yoshida K, Murata F. Nucleic acid synthesis in proliferating peroxisomes of rat liver as revealed by electron microscopical radioautography. Histochem J 1982; 14: 197-204. http://dx.doi.org/10.1007/BF01041214
Nagata T, Fujii Y, Usuda N. Demonstration of extranuclear nucleic acid synthesis in mammalian cells under experimental conditions by electron microscopic radioautography. Proc. 10th Internat. Cong. Electron Microsc, Hamburg, Germany 1982; Vol. 2: pp. 305-306.
Ma H, Nagata T. Studies on DNA synthesis of aging mice by means of electron microscopic radioautography. J Clin Electron Microsc 1988; 21: 335-43.
Ma H, Nagata T. Electron microscopic radioautographic study of DNA synthesis in the livers of aging mice. J Clin Electron Microsc 1988; 21: 715-16.
Ma H, Gao F, Sun L, Jin C, Nagata T. Electron microscopic radioautographic study on the synthesis of DNA, RNA and protein in the livers of aging mice. Med Electron Microsc 1994; 27: 349-51. http://dx.doi.org/10.1007/BF02349694
Nagata T. Light and electron microscopic radioautographic studies on macromolecular synthesis in amitotic hepatocytes of aging mice. Cell Mol Biol 2003; 49: 591-11.
Nagata T, Ma H. Electron microscopic radioautographic study on mitochondrial DNA synthesis in hepatocytes of aging mouse. Ann Microsc 2005; 5: 4-18.
Nagata T. Electron microscopic radioautographic study on protein synthesis in hepatocyte mitochondria of developing mice. Ann Microsc 2006; 6: 42-54.
Nagata T. Electron microscopic radioautographic study on macromolecular synthesis in hepatocyte mitochondria of aging mouse. J Cell Tissue Res 2007; 7: 1019-29.
Nagata T. Electron microscopic radioautographic study on nucleic acids synthesis in hepatocyte mitochondria of developing mice. Trends Cell Mol Biol 2007; 2: 19-33.
Nagata T. Electron microscopic radioautographic study on protein synthesis in mitochondria of binucleate hepatocytes in aging mice. Scientific World J 2007; 7: 1008-23. http://dx.doi.org/10.1100/tsw.2007.140
Nagata T, Usuda N, Ma H. Electron microscopic radioautography of nucleic acid synthesis in pancreatic acinar cells of prenatal and postnatal aging mice. Proc. 11th Internat. Cong. Electron Microsc, Kyoto, Japan 1986; Vol. 3, pp. 2281-2282.
Sun L, Gao F, Jin C, Nagata T. DNA synthesis in the trachea of aging mice by light and electron microscopic radioautography. Acta Histochem Cytochem 1997; 30: 211-20. http://dx.doi.org/10.1267/ahc.30.211
Nagata T, Sun L. Electron Microscopic Radioautographic Study on Mitochondrial DNA and RNA Syntheses in Pulmonary Cells of Aging Mice. Ann Microsc 2007; 7: 36-59.
Hanai T, Nagata T. Electron microscopic radioautographic study on nucleic acid synthesis in perinatal mouse kidney tissue. Med Electron Microsc 1994; 27: 355-57. http://dx.doi.org/10.1007/BF02349695
Gao F, Ma H, Sun L, Jin C, Nagata T. Electron microscopic radioautographic study on the nucleic acid and protein synthesis in the aging mouse testis. Med Electron Microsc 1994; 27: 360-62. http://dx.doi.org/10.1007/BF02349697
Gao F, Chen S, Sun L, Kang W, Wang Z, Nagata T. Radioautographic study of the macromolecular synthesis of Leydig cells in aging mouse testis. Cell Mol Biol 1995; 41: 145-50.
Yamada AT, Nagata T. Light and electron microscopic radioautography of DNA synthesis in the endometria of pregnant ovariectomized mice during activation of implantation window. Cell Mol Biol 1992; 38: 763-74.
Yamada AT, Nagata T. Light and electron microscopic radioautographic studies on the RNA synthesis of peri-implanting pregnant mouse uterus during activation of receptivity for blastocyst implantation. Cell Mol Biol 1993; 39: 221-33.
Ito M, Nagata T. Electron microscopic radioautographic study on DNA synthesis and the ultrastructure of the adrenal gland in aging mice. Med Electron Microsc 1996; 29: 145-52. http://dx.doi.org/10.1007/BF02348068
Ito M. Radioautographic studies on aging changes of DNA synthesis and the ultrastructural development of mouse adrenal gland. Cell Mol Biol 1996; 42: 279-92.
Nagata T. Electron microscopic radioautographic study on mitochondrial DNA synthesis in adrenal cortical cells of developing mice. J Cell Tis Res 2008; 8: 1303-12.
Cui H, Gao F, Ma H, Nagata T. Study on DNA synthesis of cellular elements in the cerebella of aging mice by light and electron microscopic radioautography. Proc. 4th China-Japan Joint Histochem. Cytochem. Symp, Chongqing Publishing House, Chongqing, China 1996; pp. 111-112.
Gunarso W. Radioautographic studies on the nucleic acid synthesis in the retina of chicken embryo II. Electron microscopic radioautography. Shinshu Med J 1984; 32: 241-48.
Gunarso W, Gao F, Cui H, Ma H, Nagata T. A light and electron microscopic radioautographic study on RNA synthesis in the retina of chick embryo. Acta Histochem 1996; 98: 300-22. http://dx.doi.org/10.1016/S0065-1281(96)80024-9
Gunarso W, Gao F, Nagata T. Development and DNA synthesis in the retina of chick embryo observed by light and electron microscopic radioautography. Cell Mol Biol 1997; 43: 189-201.
Kong Y, Nagata T. Electron microscopic radioautographic study on nucleic acid synthesis of perinatal mouse retina. Med Electron Microsc 1994; 27: 366-68. http://dx.doi.org/10.1007/BF02349699
Nagata T. Aging changes of macromolecular synthesis in the avian and mammalian eyes as revealed by microscopic radioautography. Ann Rev Biomed Sci 2006; 8: 33-67.
Korr H, Phillipi V, Helg C, Schiefer J, Graeber MB, Kreutzberg GW. Unscheduled DNA synthesis and mitochondrial DNA synthetic rate following injuring of the facial nerve. Acta Neuropathol 1997; 94: 557-66. http://dx.doi.org/10.1007/s004010050750
Korr H, Kurz C, Seidler TO, Sommer D, Schmitz C. Mitochondrial DNA synthesis studied autoradiographically in various cell types in vivo. Braz J Med Biol Res 1998; 31: 289-98. http://dx.doi.org/10.1590/S0100-879X1998000200012
Schmitz C, Axmacher B, Zunker U, Korr H. Age related changes of DNA repair and mitochondrial DNA synthesis in the mouse brain. Acta Neuropathol 1999; 97: 71-81. http://dx.doi.org/10.1007/s004010050957
Schmitz C, Materne S, Korr H. Cell-type-specific differences in age-related changes of DNA repair in the mouse brain - Molecular basis for a new approach to understand the selective neuronal vulnerability in Alzheimer’s disease. J Alzheimer’s Disease 1999; 1: 387-407.
Nagata T. Recent studies on macromolecular synthesis labeled with 3H-thymidine in various organs as revealed by electron microscopic radioautography. Curr Radiopharm 2009; 2: 118-28. http://dx.doi.org/10.2174/1874471010902020118
Nagata T. Electron microscopic radioatuographic studies on macromolecular synthesis in mitochondria of various cells. 18EMSM Conference Proc. 9th Asia-Pacific Microscopy Conference (APMC9), Kuala Lumpur, Malaysia 2009; pp. 48-50.
Nagata T. Electron microscopic radioautographic studies on macromolecular synthesis in mitochondria of animal cells in aging. Ann Rev Biomed Sci 2009; 11: 1-17.
Nagata T. Electron microscopic radioautographic studies on macromoleclular synthesis in mitochondria of some organs in aging animals. Bull Shinshu Inst Alternat Med Welfare 2009; 4: 15-38.
Nagata T. Electron microscopic radioautographic studies on macromolecular synthesis in mitochondria ofanimal cells in aging. Ann Rev Biomed Sci 2010; 12: 1-29.
Nagata T. Macromolecular synthesis in the livers of aging mice as revealed by electron microscopic radioautography. In, Progress in Histochemistry and Cytochemistry. Vol. 45, No. 1, pp. 1-80, Elsevier, Jena, New York, 2010
Nagata T. DNA synthesis in the pancreatic acinar cells of aging mice as revealed by electron microscopic radioautography. IIOABJ (Institute of Integrative Omics and Applied Biotechnology) 2011; 2: 31-39.
Nagata T. RNA Synthesis in the pancreatic acinar cells of aging mice as revealedby electron microscopic radioautography. Curr Radiophamaceutics 2012; 5: 5-14. http://dx.doi.org/10.2174/1874471011205010005
