Summary
Using histochemical procedures for the detection of lactate dehydrogenase (LDH), succinate dehydrogenase (SDH), and cytochrome c oxidase (cytox), we investigated the levels of these enzymes of the energy metabolism in postimplantation rat embryos (9.5–12.5 days of gestation). On day 10.5 of gestation, the neural tube, somites, myocardium, and mesenchyme displayed moderate levels of LDH activity; this activity gradually increased in strength, so that, on day 12.5 of gestation, intense LDH activity was uniformly distributed in these intraembryonic tissues. In contrast to LDH, distinet regional differences in the distribution of SDH and cytox were detected. On day 10.5 of gestation, the myocardium exhibited weak to moderate SDH and cytox activity, and on day 11.5, the myocardial activity of these enzymes had become moderate to intense. However, in all other embryonic tissues, e.g., the neural tube and somites, only weak SDH and cytox activity was present. On day 12.5 of gestation, the myocardium displayed very intense SDH and cytox activity, whereas the mantle layer of the neural tube, the spinal ganglia, and the myotomes exhibited only moderate levels of SDH and cytox activity. In the matrix of the neural tube and mesenchyme, these enzyme activities remained at low levels. At electron microscopy, cytox activity was detectable in the spaces between the inner and outer membranes as well as in the intracristal spaces of mitochondria. In general, cytox activity increased in paralled with the differentiation of mitochondria (i.e., increased mitochondrial numbers and size, and the development of mitochondrial cristae), but when the distribution of the cytox activity was considered in detail, it was found to differ among mitochondria. The relationship between, on the one hand, changes in the enzymatic patterns with a bearing on the energy-yielding metabolism and, on the other hand, cellular differentiation during major organogenesis in rat embryos is discussed.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Aksu O, Mackler B, Shepard TH, Lemire RJ (1968) Studies of the development of congenital anomalies in embryos of riboflavin-deficient, galactoflavin fed rats. II. Role of the terminal electron transport systems. Teratology 1:93–102
Altman J, Bayer SA (1984) The development of the rat spinal cord. Adv Anat Embryol Cell Biol 85:1–164
Anderson WA, Perotti ME (1975) An ultracytochemical study of the respiratory potency, integrity, and fate of the sea urchin sperm mitochrondria during early embryogenesis. J Cell Biol 66:367–376
Anderson WA, Bara G, Seligman AM (1975) The ultrastructural localization of cytochrome oxidase via cytochrome c1. J Histochem Cytochem 23:13–20
Angermüller S, Fahimi HD (1981) Selective cytochemical localization of peroxidase, cytochrome oxidase and catalase in rat liver with 3,3′-diaminobenzidine. Histochemistry 71:33–44
Brown NA, Fabro S (1981) Quantitation of rat embryonic development in vitro: A morphological scoring system. Teratology 24:65–78
Clough JR, Whittingham DG (1983) Metabolism of (14C)glucose by postimplantation mouse embryos in vitro. J Embryol Exp Morphol 74:133–142
Cockroft DL (1979) Nutrient requirements of rat embryo's undergoing organogenesis in vitro. J Reprod Fertil 57:505–510
Gass P, Kugler P (1984) Enzymdifferenzierung des visceralen Dottersackepithels der Ratte. Z Mikrosk Anat Forsch 98:721–745
Harper HA (1969) Review of physiological chemistry. 13th Maruzen Asian Edition. Maruzen, Tokyo, Japan
Hirai K-I, Ogawa K (1986) Cytochemical quantitation of cytochrome oxidase activity in rat pulmonary alveolar epithelial cells and possible defect in type I cells. J Electron Microsc 35:19–28
Jakoveic S, Haddock J, Getz GS, Rabinowitz M, Swift H (1971) Mitochondrial development in liver of foetal and newborn rats. Biochem J 121:341–347
Lewis PR, Knight DP (1977) Staining methods for sectioned material. North-Holland, Amsterdam New York Oxford, pp 25–53
Loida Z, Gossrau R, Schiebler TH (1979) Enzyme histochemistry: A laboratory manual. Springer, Berlin Heidelberg New York
Mackler B, Grace R, Duncan HM (1971) Studies of mitochondrial development during embryogenesis in the rat. Arch Biochem Biophys 144:603–610
Miki A, Kugler P (1984) Comparative enzyme histochemical study on the visceral yolk sac endoderm in the rat in vivo and in vitro. Histochemistry 81:409–415
Miki A, Mizoguti H (1982a) Relationship between the proliferating ability and acetylcholinesterase activity of the myotomal cells in the early chick embryo. Acta Histochem Cytochem 15:474
Miki A, Mizoguti H (1982b) Acetylcholinesterase activity in the myotome of the early chick embryo. Cell Tissue Res 227:23–40
Miki A, Mizoguti H (1986) Orgin of myotome cells in the chick embryo studied by3H-thymidine autoradiography and acetylcholinesterase histochemistry. Acta Histochem Cytochem 19:11–20
New DAT (1978) Whole-embryo culture and the study of mammalian embryos during organogenesis. Biol Rev 53:81–122
Nilsson BO, Magnusson C, Widehn S, Hillensjo T (1982) Correlation between blastcyst oxygent consumption and trophoblast cytochrome oxidase reaction at initiation of implantation of delayed mouse blastcysts. J Embryol Exp Morphol 71:75–82
Schoenen J (1978) Histoenzymology of the developing rat spinal cord. Neuropathol Appl Neurobiol 4:37–46
Seligman AM, Karnovsky MJ, Wasserkrug HL, Hanker JS (1968) Nondroplet ultrastructural demonstration of cytochrome oxidase activity with a polynerizing osmiophilic reagent, diaminobenzidine (DAB). J Cell Biol 38:1–14
Shepard TH, Tanimura T, Robkin MA (1970) Energy metabolism in early mammalian embryos. Dev Biol (Suppl) 4:42–68
Stern S, Biggers JD, Anderson E (1971) Mitochondria and early development of the mouse. J Exp Zool 176:179–192
Tanimura T, Shepard TH (1970) Glucose metabolism by rat embryos in vitro. Proc Soc Exp Biol Med 135:51–54
Venable JH, Coggeshall R (1965) A simplified lead citrate stain for use in electron microscopy. J Cell Biol 25:407–408
Vertel BM, Fischman DA (1977) Mitochondrial development during myogenesis. Dev Biol 58:356–371
Author information
Authors and Affiliations
Additional information
Dedicated to Professor Dr. T.H. Schiebler on the occasion of his 65th birthday
Rights and permissions
About this article
Cite this article
Miki, A., Mizoguchi, A. & Mizoguti, H. Histochemical studies of enzymes of the energy metabolism in postimplantation rat embryos. Histochemistry 88, 489–495 (1988). https://doi.org/10.1007/BF00570314
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00570314