To determine the extent to which elevated glucose and 3-hydroxybutyrate (3OHB) concentrations contribute to the embryotoxic properties of diabetic serum, we tested the effects of serum from untreated or acutely insulin-treated diabetic rats on the development of mouse embryos during neurulation in vitro. Male Sprague-Dawley rats (n = 143) with streptozocin-induced diabetes for 1 week received infusions of insulin (n = 105) or saline (n = 38) for up to 120 min. The insulin-infused animals were exsanguinated when serum glucose concentrations fell to between 5.6 and 8.3 mM. Saline-infused animals were exsanguinated after a similar duration of infusion. Serum samples were tested for embryotoxic effects on 3–6 somite mouse embryos cultured in vitro for 24 h. Of embryos cultured in serum from untreated diabetic animals (glucose: 24 ± 1 mM; 3OHB: 2.0 ± 0.3 mM), 36% (31 of 87) exhibited gross malformations, mostly of the neural tube. Only 16% (10 of 62) of embryos grown in serum from acutely insulin-treated animals (glucose: 7.4 ± 0.2 mM; 3OHB: 0.20 ± 0.06 mM) were malformed. This rate that was < half the rate caused by exposure to diabetic serum (P < 0.01), but a rate that remained much > the rate associated with culture in normal serum (0% in this study; < 2% historically). In vitro addition of glucose to serum from insulin-treated animals to re-establish hyperglycemia in the diabetic range (25 mM) resulted in a 17% (12 of 70) malformation rate, nearly identical to the 16% rate caused by normoglycemic serum from insulin-treated animals. Addition of D-3OHB to approximate the highest concentration encountered in untreated diabetic serum (∼4 mM) increased the malformation rate to 29% (21 of 73 embryos; P = 0.08 vs. unmodified serum from insulin-treated animals). Addition of glucose (to ∼25 mM) and 3OHB (to ∼4 mM) resulted in a 23% malformation rate, which was not significantly different from the rate caused by unmodified serum from the insulintreated group. These findings demonstrate that factors other than glucose and 3OHB account for much of the teratogenic potential of diabetic serum in this in vitro culture system. Moreover, the data suggest that glucose perse is not a major cause of the embryotoxicity of diabetic serum in this system, whereas 3OHB may account for some of the toxic effects.
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May 01 1994
Diabetic Teratogenesis: In Vitro Evidence for a Multifactorial Etiology With Little Contribution From Glucose Per Se
Thomas A Buchanan;
Thomas A Buchanan
Departments of Medicine and Obstetrics and Gynecology, University of Southern California School of Medicine
Los Angeles, California
Department of Cell Biology and Anatomy, University of North Carolina
Chapel Hill, North Carolina
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Kelly M Denno;
Kelly M Denno
Departments of Medicine and Obstetrics and Gynecology, University of Southern California School of Medicine
Los Angeles, California
Department of Cell Biology and Anatomy, University of North Carolina
Chapel Hill, North Carolina
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George F Sipos;
George F Sipos
Departments of Medicine and Obstetrics and Gynecology, University of Southern California School of Medicine
Los Angeles, California
Department of Cell Biology and Anatomy, University of North Carolina
Chapel Hill, North Carolina
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Thomas W Sadler
Thomas W Sadler
Departments of Medicine and Obstetrics and Gynecology, University of Southern California School of Medicine
Los Angeles, California
Department of Cell Biology and Anatomy, University of North Carolina
Chapel Hill, North Carolina
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Address correspondence and reprint requests to Dr. Thomas A. Buchanan, Room 8250 General Hospital, 1200 North State Street, Los Angeles, CA 90033.
Diabetes 1994;43(5):656–660
Article history
Received:
April 20 1993
Revision Received:
December 23 1993
Accepted:
December 23 1993
PubMed:
8168642
Citation
Thomas A Buchanan, Kelly M Denno, George F Sipos, Thomas W Sadler; Diabetic Teratogenesis: In Vitro Evidence for a Multifactorial Etiology With Little Contribution From Glucose Per Se. Diabetes 1 May 1994; 43 (5): 656–660. https://doi.org/10.2337/diab.43.5.656
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