BMP7 is Downregulated in Lumbosacral Spinal Cord of Rat Embryos With Anorectal Malformation

Abstract

BackgroundBone morphogenetic proteins (BMPs) comprise a highly conserved signaling protein family, which are involved in spinal cord formation, development and differentiation. Malformations of the lumbosacral spinal cord are associated with postoperation complications of anorectal malformation (ARM). However, the mechanism underlying the development of these malformations remains elusive. Materials and methodsEmbryonic rat ARM model induced by ethylenethiourea (ETU) was introduced to investigate BMP7 expression in lumbosacral spinal cord. BMP7 expression was analyzed by immunohistochemical staining, qRT-PCR, and Western blot analysis on embryonic (E) days 16, 17, 19, and 21. The expression of the neuronal marker neurofilament (NF) and pSmad1/5 was determined by immunofluorescence double staining and Western blot analysis during peak BMP7 expression. ResultsBMP7 mRNA (E16, 1.041 ± 0.169 versus 0.758 ± 0.0423, P < 0.05; E17, 1.889 ± 0.444 versus 1.601 ± 0.263, P < 0.05; E19, 2.898 ± 0.434 versus 1.981 ± 0.068, P < 0.01; and E21, 2.652 ± 0.637 versus 1.957 ± 0.09, P < 0.05;) and protein (E16, 1.068 ± 0.065 versus 0.828 ± 0.066, P < 0.01; E17, 1.728 ± 0.153 versus1.4 ± 0.148, P < 0.05; E19, 2.313 ± 0.141 versus 1.696 ± 0.21, P < 0.01; and E21, 2.021 ± 0.13 versus 1.43 ± 0.128, P < 0.01) were downregulated, and their expressions were specifically low in interneurons (IN) located in the dorsal horn of the lumbosacral spinal cord in embryos with ARM. On E19, Western blot analysis revealed reduced P-Smad1/5(1.13 ± 0.08 versus 0.525 ± 0.06, P < 0.01). ConclusionsAn implication of this study is the possibility that BMP7 downregulation contributes to maldevelopment of the lumbosacral spinal cord during embryogenesis in fetal rats with ARM, indicating that BMP7 may play an important role in ARM pathogenesis and the complications thereof.