Remodeling of the Vertebral Body in Hereditary Lordoscoliotic Rabbits Revealed by In Situ Hybridization

The etiology of human idiopathic scoliosis remains still unknown, although several etiologic factors and many animal models of scoliosis have been reported. Kin previously reported radiological and histological findings of spinal deformities in the Hereditary Lordoscoliotic Rabbit (LSR), which develops thoracic lordosis or lordoscoliosis during growth. We investigated biological activities of bone cells of the LSR vertebrae by in situ hybridization. Digoxigenin labeled in situ hybridization probes of mRNA of osteopontin, type I collagen and bone morphogenetic protein (BMP)-2 were prepared. Hybridization was carried out on the sagittal section of lordotic or lordoscoliotic thoracic vertebra in the LSR. In the apical vertebral body and spinous process of the lordotic thoracic spine of LSR during developmental age, stronger positive signals for type I collagen and BMP-2 mRNA were detected in osteoblasts in the endosteum of the ventral portion than those in the dorsal portion of the vertebra, which indicates osteogenesis. In the apical vertebral body of the lordotic thoracic spine, positive signals for osteopontin mRNA were detected in osteoclasts of the Howship’s lacunae and osteocytes around these pits in dorsal endosteal portion, which indicates bone resorption. No such signals were detected in the ventral endosteal portion of the same vertebral body. The vertebral bones of the LSR showed a specific mode of remodeling response to the force producing lordosis

Spontaneous Establishment of a Novel Human Cell Line Derived From Lens Epithelium

This is the first report on the spontaneous establishment of a human lens epithelial cell line. The cell line obtained from a surgical specimen of cataract was established in vitro, and subcultured for three years. The cells were spindle/dendrite-shaped, mitochondria-rich, and morphologically similar to lens epithelial cells in in vitro primary culture. The karyotype of the cells was 45,X, and the nucleotide sequence of the myoglobin gene was completely identical to that of the same gene in humans. The cell line was free from contamination with microbes such as mycoplasma. The production ofαA crystalline by the cell line was demonstrated by immunohistochemistry. The production of type I and type IV collagens,αA crystallin andα-smooth muscle actin (αSMA) by the cell line was increased by the stimulation of transforming growth factor (TGF)β1 andβ2. In addition, since we were able to show the inhibition of the production of the above proteins by anti-after-cataract agents, the cell line is considered to be a beneficial tool in research for developing therapeutic drugs for after-cataract and other diseases involving lens epithelial cells. The cell line stimulated by TGF βs may be useful as an in vitro model of lens disease

Post-weaning increases in the milk-fat globule EGF-factor VIII on fat globules in mouse milk and in the uptake of the fat globules by HC11 mammary epithelial cells

Milk fat globules (MFGs) secreted by lactating mammary gland are unique lipid surrounded by a phospholipid bi-layer. We report here post-weaning changes in milk fat globule EGF factor VIII (MFG-E8) and annexin V-accessible phosphatidylserine on the surface of MFGs. The MFG content in milk markedly decreased to about half within 2 days after forced weaning, despite a slight increase in milk protein content. Immunofluorescence-staining of MFGs using anti-MFG-E8 and annexin V indicated that MFG-E8 was present on some, but not all, MFGs before weaning, whereas most of MFGs were MFG-E8-positive and annexin V-negative after weaning. Free MFG-E8 with binding activity to phosphatidylserine was present abundantly in the post-weaning milk, and indeed exhibited binding to MFGs in pre-weaning milk. MFGs were taken up by HC11 mouse mammary epithelial cells in vitro, and those from post-weaning milk were remarkable for such cellular uptake. Moreover, the uptake of MFGs by the cells was inhibited by an anti-MFG-E8 antibody. Taken together, these findings suggest that MFG-E8 plays a critical role in regulation of MFG dynamics after weaning or during the suckling interval through the control of MFG-epithelial cell interaction in lactating mammary glands