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Biotin Influences Palatal Development of Mouse Embryos in Organ Culture^1,2,

Department of Biochemistry and Molecular Biology ^* Department of Anatomy, Faculty of Medicine, University of Manitoba, Winnipeg, Manitoba, Canada R3E 0W3

Maternal biotin deficiency is strongly teratogenic in CD-1 mice. The most common malformations are craniofacial and limb defects such as cleft palate, micrognathia and micromelia. The effect of biotin deficiency on palatal development in mouse embryos on d 12 of gestation was studied by culturing mouse embryonic palates in serum-free medium using a suspension culture system. In control embryos palatal processes developed to the fused stage after 72 h in culture. The fusion of palatal processes was further increased by the addition of biotin (10^-8 mol/L) to the medium. The addition of organic acids such as propionic, ß-methyl crotonic or ß-hydroxy isovaleric acids as well as avidin to the medium did not affect the stage of palatal formation. Cycloheximide completely blocked the fusion of palatal shelves. In embryos from biotin-deficient mice, the incidence of fusion between the palatal shelves was <7% and increased to >30% when biotin (10^-8-10^-6 mol/L) was added to the medium. The addition of fatty acids to the organ culture medium did not have any effect on the fusion of palatal processes. The incorporation of ³⁵S-methionine into protein from biotin-deficient embryo explants was 88% of that in controls. The results indicate that biotin deficiency may interfere directly with synthesis of specific proteins and the formation of palatal processes.

KEY WORDS: • biotin deficiency • palatal development • organ culture • teratogenicity • mice

¹ Supported by a grant from the Medical Research Council of Canada. T.W. held a Medical Research Council Visiting Scientist Award.

² The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 USC section 1734 solely to indicate this fact.

³ Current address: Department of Hygiene and Preventive Medicine, Yamagata University School of Medicine, Yamagata 99023, Japan.

⁴ To whom correspondence and reprint requests should be addressed.

Manuscript received 5 July 1994. Revision accepted 7 March 1995.