Summary
To demonstrate the myo-inositol depletion hypothesis in hyperglycaemia-induced embryopathy, rat conceptuses of 9.5 days of gestation in the early head-fold stage were grown in vitro during neural tube formation for 48 h with increasing amounts of glucose. The effects of an aldose reductase inhibitor and the myo-inositol supplementation were also investigated. Sorbitol and myo-inositol contents were measured in separated embryos and extra-embryonic membranes including yolk sac and amnion at the end of culture. After addition of 33.3 mmol/l and 66.7 mmol/l glucose to the culture media, the myo-inositol content of the embryos was significantly decreased by 43.1% (p<0.05) and 64.6% (p < 0.01) of the control group, while a marked accumulation of sorbitol was observed (25 and 41 times that of the control). Although the addition of an aldose reductase inhibitor (0.7 mmol/l) to the hyperglycaemic culture media containing an additional 66.7 mmol/l glucose significantly reduced the sorbitol content of embryos to approximately one-eighth, the myo-inositol content of embryos remained decreased and the frequency of neural lesions was unchanged (23.1% vs 23.9%, NS). Supplementation of the myo-inositol (0.28 mmol/l) completely restored the myo-inositol content of the embryos and resulted in a significant decrease in the frequency of neural lesions (7.1% vs 23.9%, p < 0.01) and a significant increase in crown-rump length and somite numbers. Much less significantly, sorbitol accumulation was also observed in the extra-embryonic membrane in response to hyperglycaemia, neither hyperglycaemia nor the myo-inositol supplementation modified the myo-inositol contents of the extra-embryonic membrane. We conclude that the mechanism of hyperglycaemia-induced teratogenicity was mediated by the myo-inositol depletion of the embryo at a critical stage of organogenesis.
Article PDF
Similar content being viewed by others
References
Mills JL (1982) Malformations in infants of diabetic mothers. Teratology 25: 385–394
Small M, Cassidy L, Leiper JM, Paterson KR, Lunan CB, MacCuish AC (1986) Outcome of pregnancy in insulin-dependent (Type I) diabetic women between 1971 and 1984. Q J Med 61: 1159–1169
Nelson RL (1986) Diabetes and pregnancy: control can make a difference. Mayo Clin Proc 61: 825–829
Mills JL, Baker L, Goldman AS (1979) Malformations in infants of diabetic mothers occur before the seventh gestational week. Implications for treatment. Diabetes 28: 292–293
New DAT (1978) Whole embryo culture and the study of mammalian embryos during organogenesis. Biol Rev 53: 81–122
Freinkel N, Cockroft DL, Lewis NJ, Gorman L, Akazawa S, Phillips LS, Shambaugh GE (1986) The 1986 McCollum award lecture. Fuel-mediated teratogenesis during early organogenesis: the effects of increased concentrations of glucose, ketones, or somatomedin inhibitor during rat embryo culture. Am J Clin Nutr 44: 986–995
Sadler TW (1980) Effects of maternal diabetes on early embryogenesis. II. Hyperglycemia-induced exencephaly. Teratology 21: 349–356
Cockroft DL, Coppola PT (1977) Teratogenic effects of excess glucose on head-fold rat embryos in culture. Teratology 16: 141–146
Sadler TW (1980) Effects of maternal diabetes on early embryogenesis. I. The teratogenic potential of diabetic serum. Teratology 21: 339–347
Garnham EA, Beck F, Clarke CA, Stainsstreet M (1983) Effects of glucose on rat embryos in culture. Diabetologia 25: 291–295
Ellington SKL (1987) Development of rat embryos cultured in glucose-deficient media. Diabetes 36: 1372–1378
Akazawa S, Akazawa M, Hashimoto M, Yamaguchi Y, Kuriya N, Toyama K, Ueda Y, Nakanishi T, Mori T, Miyake S, Nagataki S (1987) Effects of hypoglycaemia on early embryogenesis in rat embryo organ culture. Diabetologia 30: 791–796
Horton WE, Sadler TW (1983) Effects of maternal diabetes on early embryogenesis. Alterations in morphogenesis produced by the ketone body, B-hydroxybutyrate. Diabetes 32: 610–616
Sadler TW, Phillips LS, Balkan W, Goldstein S (1986) Somatomedin inhibitors from diabetic rat serum alter growth and development of mouse embryos in culture. Diabetes 35: 861–865
Shepard TH, Tanimura T, Robkin MA (1970) Energy metabolism in early mammalian embryos. Symp Soc Dev Biol 29: 42–58
Freinkel N, Lewis NJ, Akazawa S, Gorman L, Potaczek M (1983) The honeybee syndrome: teratogenic effects of mannose during organogenesis in rat embryo culture. Trans Assoc Am Physicians 96: 44–55
Freinkel N, Lewis NJ, Akazawa S, Roth SI, Gorman L (1984) The honeybee syndrome-implications of the teratogenicity of mannose in rat-embryo culture. N Engl J Med 310: 223–230
Lewis NJ, Akazawa S, Freinkel N (1983) Teratogenesis from β-hydroxybutyrate during organogenesis in rat embryo organ culture and enhancement by subteratogenic glucose. Diabetes 32 [Suppl 1]: 11A
Sheehan EA, Beck F, Clarke CA, Stanisstreet M (1985) Effects of β-hydroxybutyrate on rat embryos grown in culture. Experientia 41: 273–275
Sadler TW, Hunter ES III, Wynn RE, Phillips LS (1989) Evidence for multifactorial origin of diabetes-induced embryopathies. Diabetes 38: 70–74
Hunter ES III, Sadler TW, Wynn RE (1987) A potential mechanism of DL-β-hydroxybutyrate-induced malformations in mouse embryos. Am J Physiol 235: 72–80
Eriksson UJ, Naeser P, Brolin SE (1986) Increased accumulation of sorbitol in offspring of manifest diabetic rats. Diabetes 35: 1356–1363
Eriksson UJ, Brolin SE, Naeser P (1989) Influence of sorbitol accumulation on growth and development of embryos cultured in elevated levels of glucose and fructose. Diab Res 11: 27–32
Hod M, Star S, Passonneau JV, Unterman TG, Freinkel N (1986) Effect of hyperglycemia on sorbitol and myo-inositol content of cultured rat conceptus: failure of aldose reductase inhibitors to modify myo-inositol depletion and dysmorphogenesis. Biochem Biophys Res Comm 140: 974–980
Baker L, Piddington R, Goldman AS, Dahlem S, Egler J (1986) Myoinositol (MI) and Arachidonic acid (AA) are linked in the mechanism of diabetic embryopathy. Diabetes 35 [Suppl 1]: 12A
Lowry OH, Rosenbrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin Phenol reagent. J Biol Chem 193: 265–275
Dethy JM, Callaert-Deveen B, Janssens M, Lenaers A (1984) Determination of sorbitol and galactitol at the nanogram level in biological samples by high-performance liquid chromatography. Anal Biochem 143: 119–124
Snedecor GW, Cochran WG (1980) Statistical Methods, 7th edn. Iowa State University Press, Ames, Iowa, pp 64–130
Sadler TW, Hunter ES III (1987) Hypoglycemia: how little is too much for the embryo? Am J Obstet Gynecol 157: 190–193
Sussman I, Matschinsky FM (1988) Diabetes affects sorbitol and myo-inositol levels of neuroectodermal tissue during embryogenesis in rat. Diabetes 37: 974–981
Gabbay KH (1973) The sorbitol pathway and the complications of diabetes. N Engl J Med 288: 831–836
Yeh LA, Rafford CE, Beyer TA, Hutson NJ (1986) Effects of the aldose reductase inhibitor sorbinil on the isolated cultured rat lens. Metabolism 35 [Suppl 1]: 4–9
MacGregor LC, Matschisky FM (1986) Experimental diabetes mellitus impairs the function of the retinal pigmented epithelium. Metabolism 35 [Suppl 1]: 28–34
Williamson JR, Chang K, Rowold E, Marvel J, Tomlinson M, Sherman WR, Ackerman KE, Kilo C (1986) Diabetes-induced increases in vascular permeability and change in granulation tissue levels of sorbitol, myo-inositol, chiro-inositol, and scylloinositol are prevented by sorbinil. Metabolism 35 [Suppl 1]: 41–45
Kikkawa R, Umemura K, Haneda M, Arimura T, Ebata K, Shigeta Y (1987) Evidence for existence of polyol pathway in cultured rat mesangial cells. Diabetes 36: 240–243
Finegold D, Lattimer SA, Nolle S, Bernstein M, Green DA (1983) Polyol pathway activity and myo-inositol metabolism. A suggested relationship in the pathogenesis of diabetic neuropathy. Diabetes 32: 988–992
Cohen MP (1986) Aldose reductase, glomerular metabolism, and diabetic nephropathy. Metabolism 35 [Suppl 1]: 55–59
Pinter E, Reece EA, Leranth CZ (1986) Arachidonic acid prevents hyperglycemia-associated yolk sac damage and embryopathy. Am J Obstet Gynecol 155: 691–702
Goldman AS, Baker L, Piddington R, Marx B, Herold R, Egler J (1985) Hyperglycemia-induced teratogenesis is mediated by a functional deficiency of arachidonic acid. Proc Natl Acad Sci USA 82: 8227–8231
Weigensberg M, Garcia-Palmer F, Freinkel N (1987) Competition between glucose and myo-inositol for transport in the embryo: a possible contributor to the embryopathy of hyperglycemia? Clin Res 35: 863A
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Hashimoto, M., Akazawa, S., Akazawa, M. et al. Effects of hyperglycaemia on sorbitol and myo-inositol contents of cultured embryos: treatment with aldose reductase inhibitor and myo-inositol supplementation. Diabetologia 33, 597–602 (1990). https://doi.org/10.1007/BF00400203
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00400203