Enzyme-Histochemical Demonstration of Intralobular Lymphatic Vessels in the Mouse Thymus

Intralobular lymphatic vessels in the mouse thymus were demonstrated enzyme-histochemically by combined light and electron microscopy. In sections reacting to both 5′-nucleotidase (5′-Nase) and alkaline phosphatase (ALPase), the intralobular lymphatic vessels were identified as irregularly shaped sp...

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Published inArchives of Histology and Cytology Vol. 53; no. Supplement; pp. 87 - 94
Main Author KATO, Seiji
Format Journal Article
LanguageEnglish
Published Japan International Society of Histology and Cytology 1990
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ISSN0914-9465
1349-1717
1349-1717
DOI10.1679/aohc.53.Suppl_87

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Abstract Intralobular lymphatic vessels in the mouse thymus were demonstrated enzyme-histochemically by combined light and electron microscopy. In sections reacting to both 5′-nucleotidase (5′-Nase) and alkaline phosphatase (ALPase), the intralobular lymphatic vessels were identified as irregularly shaped spaces with strong 5′-Nase activity. The lymphatic vessels were closely associated with the branches of ALPase-positive intralobular arteries and veins. The initial lymphatics, which presumably originate from the perivascular spaces, were 5′-Nase positive. The distribution and intensity of the 5′-Nase activity in the lymphatic vessels revealed by light microscopy correlated well with those by backscattered image electron microscopy. The backscattered image scanning electron microscopy of the same area as observed under a light microscope more clearly highlighted the peculiar contours of lymphatic endothelial cells. Transmission electron microscopy showed that specific reaction product of the 5′-Nase after incubation in a medium containing L-tetramisole was predominantly localized on the outer surface of the lymphatic endothelial cell membranes.
AbstractList Intralobular lymphatic vessels in the mouse thymus were demonstrated enzyme-histochemically by combined light and electron microscopy. In sections reacting to both 5'-nucleotidase (5'-Nase) and alkaline phosphatase (ALPase), the intralobular lymphatic vessels were identified as irregularly shaped spaces with strong 5'-Nase activity. The lymphatic vessels were closely associated with the branches of ALPase-positive intralobular arteries and veins. The initial lymphatics, which presumably originate from the perivascular spaces, were 5'-Nase positive. The distribution and intensity of the 5'-Nase activity in the lymphatic vessels revealed by light microscopy correlated well with those by backscattered image electron microscopy. The backscattered image scanning electron microscopy of the same area as observed under a light microscope more clearly highlighted the peculiar contours of lymphatic endothelial cells. Transmission electron microscopy showed that specific reaction product of the 5'-Nase after incubation in a medium containing L-tetramisole was predominantly localized on the outer surface of the lymphatic endothelial cell membranes.
Intralobular lymphatic vessels in the mouse thymus were demonstrated enzyme-histochemically by combined light and electron microscopy. In sections reacting to both 5'-nucleotidase (5'-Nase) and alkaline phosphatase (ALPase), the intralobular lymphatic vessels were identified as irregularly shaped spaces with strong 5'-Nase activity. The lymphatic vessels were closely associated with the branches of ALPase-positive intralobular arteries and veins. The initial lymphatics, which presumably originate from the perivascular spaces, were 5'-Nase positive. The distribution and intensity of the 5'-Nase activity in the lymphatic vessels revealed by light microscopy correlated well with those by backscattered image electron microscopy. The backscattered image scanning electron microscopy of the same area as observed under a light microscope more clearly highlighted the peculiar contours of lymphatic endothelial cells. Transmission electron microscopy showed that specific reaction product of the 5'-Nase after incubation in a medium containing L-tetramisole was predominantly localized on the outer surface of the lymphatic endothelial cell membranes.Intralobular lymphatic vessels in the mouse thymus were demonstrated enzyme-histochemically by combined light and electron microscopy. In sections reacting to both 5'-nucleotidase (5'-Nase) and alkaline phosphatase (ALPase), the intralobular lymphatic vessels were identified as irregularly shaped spaces with strong 5'-Nase activity. The lymphatic vessels were closely associated with the branches of ALPase-positive intralobular arteries and veins. The initial lymphatics, which presumably originate from the perivascular spaces, were 5'-Nase positive. The distribution and intensity of the 5'-Nase activity in the lymphatic vessels revealed by light microscopy correlated well with those by backscattered image electron microscopy. The backscattered image scanning electron microscopy of the same area as observed under a light microscope more clearly highlighted the peculiar contours of lymphatic endothelial cells. Transmission electron microscopy showed that specific reaction product of the 5'-Nase after incubation in a medium containing L-tetramisole was predominantly localized on the outer surface of the lymphatic endothelial cell membranes.
Author KATO, Seiji
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References HEUSERMANN, U.: Morphologie der Lymphgefäße, der Nerven, der Kapsel und der Trabekel der menschlichen Milz. Habilischr. Med. Fakultät, Kiel, 1979 (quoted from WERNER et al., 1987).
OTSUKI, Y., Y. ITO and S. MAGARI: Lymphocyte subpopulations in high endothelial venules and lymphatic capillaries of bronchus-associated lymphoid tissue (BALT) in the rat. Amer. J. Anat. 184: 139-146 (1989).
KATO, S.: Intralobular lymphatic vessels and their relationship to blood vessels in the mouse thymus. Lightand electron-microscopy. Cell Tiss. Res. 253: 181-187 (1988).
EDWARDS, N. L., E. W. GELFAND, L. BURK, H. -M. DOSCH and I. H. Fox: Distribution of 5′-nucleotidase in human lymphoid tissues. Proc. Nat. Acad. Sci. 76: 3474-3476 (1979).
KOTANI, M., KAWAKITA, M. FUKANOGI, A. YAMASHITA, K. SEIKI and I. HORII: The passage of thymic lymphocytes to the circulation in the rat. Okajimas Fol. Anat. Jap. 43: 61-71 (1967).
MUELLER-HERMELINK, H. K.: Characterization of the B-cell and T-cell regions of human lymphatic tissue through enzyme histochemical demonstration of ATP ase and 5′-nucleotidase activities. Virchows. Arch. (B) Cell Pathol. 16: 371-378 (1974).
BURSTONE, M. S.: Enzyme histochemistry and its application in the study of neoplasms. Phosphatase. Academic Press, New York, 1962 (p. 160-292).
KATO, S.: Enzyme-histochemical identification of lymphatic vessels by light and backscattered image scanning electron microscopy. Stain Technol. 65 (1990, in press).
KATO, S. and G. I. SCHOEFL: The microvasculature of the normal and involuted mouse thymus: Light and electron microscopic study. Acta Anat. 135: 1-11 (1989).
KATO, S. and R. MIYAUCHI: Enzyme-histochemical visualization of lymphatic capillaries in the mouse tongue: Light and electron microscopic study. Okajimas Fol. Anat. Jap. 65: 391-404 (1989).
USHIKI, T.: A scanning electron-microscopic study of the rat thymus with special reference to cell types and migration of lymphocytes into the general circulation. Cell Tiss. Res. 244: 285-298 (1986).
UUSITALO, R. J. and M. J. KARNOVSKY: Surface localization of 5′-nucleotidase on the mouse lymphocytes. J. Histochem. Cytochem. 25: 87-96 (1977).
ROBINSON, J. M. and M. J. KARNOVSKY: Ultrastructural localization of 5′-nucleotidase in guinea pig neutrophils based upon the use of cerium as capturing agent. J. Histochem. Cytochem. 31: 1190-1196 (1983).
VETTER, W.: Alkaline Phosphatasen in Mastzellen, Blutund Lymphgefäßen der Rattenzunge. 5′-Nucleotidase-, unspezifische alkalische Phosphatase- und Polyphosphatase-(ATP′ase) Aktivität unter besondere Beriücksichtigung des pH. Z. Anat. Entw. -Gesch. 130: 153-176 (1970).
SMITH, C.: Studies on the thymus of the mammal. VIII. Intrathymic lymphatic vessels. Anat. Rec. 122: 173-179 (1955).
WERNER, J. A., M. SCHÜNKE and B. TILLMANN: Histochemical visualization of lymphatic capillaries in the rat: A comparison of methods demonstrated at the posterior pharyngeal surface. Arch Histol. Jap. 50: 505-514 (1987).
OMORI, H.: Finer distribution of the lymph vessels in the thymus. (In Japanese with English abstract) Acta Anat. Nippon. 48: 315-329 (1973).
HWANG, W. S., T. Y. HO, S. C. LUK and G. T. SIMON: Ultrastructure of the rat thymus. A transmission scanning electron microscope, and morphometric study. Lab. Invest. 31: 473-487 (1974).
INOUÉ, T. and H. OSATAKE: A new drying method of biological specimens for scanning electron microscopy. The t-butyl alcohol freeze-drying method. Arch. Histol. Cytol. 51: 53-59 (1988).
OTSUKI, Y. and S. MAGARI: Lymphocyte subpopulations in high endothelial venules and lymphatic capillaries in rat Peyer's patches: An immunoelectron microscopic study. Acta Histochem. Cytochem. 21: 33-44 (1988).
DE PIEREE, J. W. and M. L. KARNOVSKY: Ecto-enzyme of granulocytes: 5′-Nucleotidase. Science 184: 1096-1098 (1974).
KATO, S.: Enzyme-histochemical identification of lymphatic vessels. (In Japanese with English abstract). Jap. J. Lymphol. 12: 1-9 (1989).
WACHSTEIN, M. and E. MEISEL: Histochemistry of hepatic phosphatases at a physiologic pH. Amer. J. Clin. Pathol. 130: 153-176 (1957).
KOTANI, M., S. SEIKI, A. YAMASHITA and I. HORII: Lymphatic drainage of thymocytes to the circulation in the guinea pig. Blood 27: 511-520 (1966).
WHEELER, E. E., J. B. GAVIN and R. N. SEELYE: Freezedrying from tertiary butanol in the preparation of endothelium for scanning electron microscopy. Stain Technol. 50: 331-337 (1975).
References_xml – reference: BURSTONE, M. S.: Enzyme histochemistry and its application in the study of neoplasms. Phosphatase. Academic Press, New York, 1962 (p. 160-292).
– reference: KATO, S.: Intralobular lymphatic vessels and their relationship to blood vessels in the mouse thymus. Lightand electron-microscopy. Cell Tiss. Res. 253: 181-187 (1988).
– reference: WACHSTEIN, M. and E. MEISEL: Histochemistry of hepatic phosphatases at a physiologic pH. Amer. J. Clin. Pathol. 130: 153-176 (1957).
– reference: EDWARDS, N. L., E. W. GELFAND, L. BURK, H. -M. DOSCH and I. H. Fox: Distribution of 5′-nucleotidase in human lymphoid tissues. Proc. Nat. Acad. Sci. 76: 3474-3476 (1979).
– reference: DE PIEREE, J. W. and M. L. KARNOVSKY: Ecto-enzyme of granulocytes: 5′-Nucleotidase. Science 184: 1096-1098 (1974).
– reference: OTSUKI, Y. and S. MAGARI: Lymphocyte subpopulations in high endothelial venules and lymphatic capillaries in rat Peyer's patches: An immunoelectron microscopic study. Acta Histochem. Cytochem. 21: 33-44 (1988).
– reference: KATO, S.: Enzyme-histochemical identification of lymphatic vessels. (In Japanese with English abstract). Jap. J. Lymphol. 12: 1-9 (1989).
– reference: VETTER, W.: Alkaline Phosphatasen in Mastzellen, Blutund Lymphgefäßen der Rattenzunge. 5′-Nucleotidase-, unspezifische alkalische Phosphatase- und Polyphosphatase-(ATP′ase) Aktivität unter besondere Beriücksichtigung des pH. Z. Anat. Entw. -Gesch. 130: 153-176 (1970).
– reference: UUSITALO, R. J. and M. J. KARNOVSKY: Surface localization of 5′-nucleotidase on the mouse lymphocytes. J. Histochem. Cytochem. 25: 87-96 (1977).
– reference: HWANG, W. S., T. Y. HO, S. C. LUK and G. T. SIMON: Ultrastructure of the rat thymus. A transmission scanning electron microscope, and morphometric study. Lab. Invest. 31: 473-487 (1974).
– reference: KATO, S. and G. I. SCHOEFL: The microvasculature of the normal and involuted mouse thymus: Light and electron microscopic study. Acta Anat. 135: 1-11 (1989).
– reference: INOUÉ, T. and H. OSATAKE: A new drying method of biological specimens for scanning electron microscopy. The t-butyl alcohol freeze-drying method. Arch. Histol. Cytol. 51: 53-59 (1988).
– reference: MUELLER-HERMELINK, H. K.: Characterization of the B-cell and T-cell regions of human lymphatic tissue through enzyme histochemical demonstration of ATP ase and 5′-nucleotidase activities. Virchows. Arch. (B) Cell Pathol. 16: 371-378 (1974).
– reference: OTSUKI, Y., Y. ITO and S. MAGARI: Lymphocyte subpopulations in high endothelial venules and lymphatic capillaries of bronchus-associated lymphoid tissue (BALT) in the rat. Amer. J. Anat. 184: 139-146 (1989).
– reference: ROBINSON, J. M. and M. J. KARNOVSKY: Ultrastructural localization of 5′-nucleotidase in guinea pig neutrophils based upon the use of cerium as capturing agent. J. Histochem. Cytochem. 31: 1190-1196 (1983).
– reference: SMITH, C.: Studies on the thymus of the mammal. VIII. Intrathymic lymphatic vessels. Anat. Rec. 122: 173-179 (1955).
– reference: KATO, S. and R. MIYAUCHI: Enzyme-histochemical visualization of lymphatic capillaries in the mouse tongue: Light and electron microscopic study. Okajimas Fol. Anat. Jap. 65: 391-404 (1989).
– reference: KATO, S.: Enzyme-histochemical identification of lymphatic vessels by light and backscattered image scanning electron microscopy. Stain Technol. 65 (1990, in press).
– reference: KOTANI, M., S. SEIKI, A. YAMASHITA and I. HORII: Lymphatic drainage of thymocytes to the circulation in the guinea pig. Blood 27: 511-520 (1966).
– reference: OMORI, H.: Finer distribution of the lymph vessels in the thymus. (In Japanese with English abstract) Acta Anat. Nippon. 48: 315-329 (1973).
– reference: WHEELER, E. E., J. B. GAVIN and R. N. SEELYE: Freezedrying from tertiary butanol in the preparation of endothelium for scanning electron microscopy. Stain Technol. 50: 331-337 (1975).
– reference: WERNER, J. A., M. SCHÜNKE and B. TILLMANN: Histochemical visualization of lymphatic capillaries in the rat: A comparison of methods demonstrated at the posterior pharyngeal surface. Arch Histol. Jap. 50: 505-514 (1987).
– reference: KOTANI, M., KAWAKITA, M. FUKANOGI, A. YAMASHITA, K. SEIKI and I. HORII: The passage of thymic lymphocytes to the circulation in the rat. Okajimas Fol. Anat. Jap. 43: 61-71 (1967).
– reference: HEUSERMANN, U.: Morphologie der Lymphgefäße, der Nerven, der Kapsel und der Trabekel der menschlichen Milz. Habilischr. Med. Fakultät, Kiel, 1979 (quoted from WERNER et al., 1987).
– reference: USHIKI, T.: A scanning electron-microscopic study of the rat thymus with special reference to cell types and migration of lymphocytes into the general circulation. Cell Tiss. Res. 244: 285-298 (1986).
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SubjectTerms 5'-Nucleotidase - analysis
Alkaline Phosphatase - analysis
Animals
Female
Histocytochemistry
Lymphatic System - anatomy & histology
Lymphatic System - enzymology
Male
Mice
Mice, Inbred BALB C
Microscopy, Electron
Microscopy, Electron, Scanning
Thymus Gland - anatomy & histology
Thymus Gland - enzymology
Title Enzyme-Histochemical Demonstration of Intralobular Lymphatic Vessels in the Mouse Thymus
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