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Contributed by Philip W. Majerus, April 20, 2009 (received for review February 11, 2009)
Article Figures & SI
Figures
- Fig. 1.
Characterization of the ITPK1 antibody and ITPK1 protein levels in mice homozygous for the Itpk1–βgal gene trap allele. (A) Western blot analysis of soluble extracts from cadaveric human and mouse brain (6 μg protein/lane). (B and C) Immunoperoxidase staining of wild-type mouse brain showing ITPK1 immunoreactivity in hippocampal (B) and brainstem (C) neurons. (Scale bar = 30 μm.) (D and E) Diagram of the gene trap allele. Alternative mRNA splicing of the gene trap allele produces both a full-length wild-type transcript (D) and Itpk1–βgal fusion transcript (E). (F) Western blot analysis of ITPK1 protein in soluble brain extracts (2.5 μg protein/lane) from wild-type mice and mice either heterozygous or homozygous for the gene trap allele. (G and H) Correlation of Xgal staining and Itpk1 mRNA expression in a mouse heterozygous for the Itpk1–βgal allele. Adjacent cryosections of hippocampus (hc) were subjected to Xgal staining (G) or in situ hybridization for Itpk1 mRNA (H; dark-field microscopy). (Scale bar = 50 μm.)
- Fig. 2.
Expression of Itpk1 in the developing mouse. (A) Whole-mount Xgal staining of a mouse embryo heterozygous for the Itpk1–βgal gene trap allele. (B–L) Tissue sections from mice heterozygous for Itpk1–βgal gene trap allele were subjected to Xgal staining. Expression of the transgene was evident in rostal neuroepithelium (ne) and retinal neurons (r) (B, inset); ganglia of cranial nerves V, VII, and VIII (C); dorsal root ganglia (drg) of the lower trunk (D); adrenal medulla (m) (E); brown adipose tissue (bat) and vascular smooth muscle cells in an adjacent artery (ar) but not in skeletal muscle (sk) (F); coronary arteries (ca) (G); airway smooth muscle (sm) (H); neurons of the lateral temporal lobe and hippocampus (hc, inset) (I); neurons in the cerebellum, particularly in the Purkinje cell layer (pl; inset) (J); vascular smooth muscle cells in arterioles (ar) of the kidney (inset shows a juxtaglomerulary arteriole) (K); and serosal cells (s), visceral smooth muscle cells (sm), and arteriolar smooth muscle cells (ar) of the duodenum (L).
- Fig. 3.
NTDs and axial skeleton malformations in mouse embryos homozygous for the Itpk1–βgal gene trap allele. (A–C) Scanning electron micrographs of a normal-appearing E11.5 mouse (A) and 2 littermates (B and C) with varying degrees of exencephaly (arrowheads). The inset in panel C shows another view of the open neural tube. (D) Xgal-stained E10.5 mouse with both exencephaly (arrowhead) and spina bifida (arrows). The dashed line indicates the orientation of the tissue section shown in the inset. The asterisk highlights the open neural tube. (E) Nascent myelomeningocele (arrowheads) in an E11.5 mouse. (F and G) Dashed lines indicate the orientation of tissue sections through the intact neural tube (F) and open neural tube (G, asterisk). (H and I) E14.5 mouse skeletons stained with alcian blue and alizarin red to visualize cartilage and bone. The inset in panel H shows 2 littermates, one of which has kyphoscoliosis (arrowhead). Higher-magnification views of this embryo are shown in panels H and I. Note the presence of malformed ribs (arrowheads in I). Abbreviations: drg, dorsal root ganglia; hl, hind limb; lb, limb bud; mc, myelocele; ntc, notochord.
Tables
E9.5 E10.5 E11.5 E12.5 Total number of embryos 36 27 69 56 % with NTD 17 11 17 4 % resorbed 0 11 12 11 % growth retarded 6 4 1 0 % total birth defects 22 26 30 14
Data supplements
Supporting Information
Files in this Data Supplement:
Neural tube defects in mice with reduced levels of inositol 1,3,4-trisphosphate 5/6-kinase
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