Keisuke Oboki

Reduced expression of IL-12 receptor β2 and IL-18 receptor α genes in natural killer cells and macrophages derived from B6-mi/mi mice

The mi transcriptional factor (MITF) is a basic helix–loop–helix leucine zipper-type transcriptional factor. The mi mutant allele encodes an abnormal MITF, in which one out of four consecutive arginines is deleted in the basic domain. The VGA-9-tg (tg) allele is another mutant allele and considered to be a null mutant allele. C57BL/6 (B6)-mi/mi mice showed abnormal phenotypes of natural killer (NK) cells and macrophages, whereas B6-tg/tg mice did not. The expression levels of the genes for the interleukin-12 receptor (IL-12R) β2 and IL-18Rα were reduced in both the NK cells and macrophages of B6-mi/mi mice, while the expression levels of the corresponding genes in B6-tg/tg mice were unaffected. The B6-mi/mi NK cells and B6-mi/mi macrophages showed impaired responses to stimulation with IL-12, IL-18, and IL-12 plus IL-18 stimulation. The abnormal NK cell and macrophage of B6-mi/mi mice appear to be due to decreased expression of the IL-12Rβ2 and IL-18Rα genes.

Number of mast cells in the peritoneal cavity of mice: Influence of microphthalmia transcription factor through transcription of newly found mast cell adhesion molecule, spermatogenic immunoglobulin superfamily

The mi (microphthalmia) locus of mice encodes a transcription factor, MITF. B6-tg/tg mice that do not express any MITF have white coats and small eyes. Moreover, the number of mast cells decreased to one-third that of normal control (+/+) mice in the skin of B6-tg/tg mice. No mast cells were detectable in the stomach, mesentery, and peritoneal cavity of B6-tg/tg mice. Cultured mast cells derived from B6-tg/tg mice do not express a mast cell adhesion molecule, spermatogenic immunoglobulin superfamily (SgIGSF). To obtain in vivo evidence for the correlation of nonexpression of SgIGSF with decrease in mast cell number, we used another MITF mutant, B6-mi(vit)/mi(vit) mice that have a mild phenotype, ie, black coat with white patches and eyes of normal size. B6-mi(vit)/mi(vit) mice had a normal number of mast cells in the skin, stomach, and mesentery, but the number of peritoneal mast cells decreased to one-sixth that of +/+ mice. Cultured mast cells and peritoneal mast cells of B6-mi(vit)/mi(vit) mice showed a reduced but apparently detectable level of SgIGSF expression, demonstrating the parallelism between mast cell number and expression level of SgIGSF. The number of peritoneal mast cells appeared to be influenced by MITF through transcription of SgIGSF.

Dual abnormal effects of mutant MITF encoded by Miwh allele on mouse mast cells: decreased but recognizable transactivation and inhibition of transactivation

MITF is a basic helix-loop-helix leucine zipper-type transcription factor and is important for development of mast cells. MITF encoded by Mi(wh) allele (Mi(wh)-MITF) was mutated at a single amino acid of basic domain, and possessed a deficient but apparent DNA-binding ability. Here, we characterized the unique effects of Mi(wh)-MITF on the expression of mast cell-related genes. The expression level of mouse mast cell protease (mMCP)-4, -5, and -6 genes in Mi(wh)/Mi(wh) cultured mast cells (CMCs) was intermediate between levels of normal (+/+) CMCs and tg/tg CMCs, which did not express any MITFs. Mi(wh)-MITF appeared to show the positive transactivation effect through the remaining DNA-binding ability. On the other hand, the expression level of tryptophan hydroxylase gene was lower in Mi(wh)/Mi(wh) CMCs than in tg/tg CMCs, suggesting the inhibitory effect of Mi(wh)-MITF on the transactivation. Mi(wh)-MITF possessed dual abnormal effects on transactivation of mast cell-related genes.

Molecular cause of the severe functional deficiency in osteoclasts by an arginine deletion in the basic domain of Mi transcription factor.

Abstract Severe osteopetrosis was observed in mi/mi mutant mice. However, the bone of VGA9/VGA9 mutant mice, in which Mi gene expression is undetectable, showed normal histology. No osteopetrosis was found in mi/+ mice, but was observed in VGA9/mi mice. Biochemical analysis revealed that the gene product encoded with the mi mutant allele (mi–Mi) has impared DNA binding activity and nuclear translocation ability. Furthermore, inhibitory effects of mi–Mi were shown not only on the DNA binding activity of wild-type Mi, but also on the nuclear translocation ability of Mi, PU.1 and cFOS. The present results suggest the presence of a target gene for Mi that is essential for the proliferation/differentiation of osteoclasts.

Deficient Eosinophil Chemotaxis-Promoting Activity of Genetically Normal Mast Cells Transplanted into Subcutaneous Tissue of Mitfmi-vga9/Mitfmi-vga9 Mice : Comparison of the Activity and Mast Cell Distribution Pattern with KitW/KitW-vMice

Despite the practical lack of mast cells in the skin tissue of WBB6F(1)-Kit(W)/Kit(W-v), the skin tissue of WBB6F(1)-Mitf(mi-vga9)/Mitf(mi-vga9) mice contains one third of mast cells than that of WBB6F(1)-+/+ mice. We attempted to investigate the function of the decreased but appreciable number of mast cells in the skin of WBB6F(1)-Mitf(mi-vga9)/Mitf(mi-vga9) mice. The substance P (SP)-induced eosinophil infiltration was examined using air-bleb assay. The air-bleb membrane was composed of the subcutaneous connective tissue. Unexpectedly, we found that the air-bleb membranes formed in the back of WBB6F(1)-Mitf(mi-vga9)/Mitf(mi-vga9) mice contained no mast cells. The WBB6F(1)-Mitf(mi-vga9)/Mitf(mi-vga9) mice showed impaired SP-induced eosinophil infiltration as observed in WBB6F(1)-Kit(W)/Kit(W-v) mice, indicating that mast cells detected in the dermis of WBB6F(1)-Mitf(mi-vga9)/Mitf(mi-vga9) mice did not help SP-induced eosinophil infiltration. Subcutaneous transplantation of cultured mast cells from WBB6F(1)-+/+ mice normalized SP-induced eosinophil infiltration in WBB6F(1)-Kit(W)/Kit(W-v) mice but not in WBB6F(1)-Mitf(mi-vga9)/Mitf(mi-vga9) mice. The greater number and the more dispersed distribution pattern of mast cells that appeared in the subcutaneous connective tissue of WBB6F(1)-Kit(W)/Kit(W-v) mice after the transplantation appeared to explain the difference between WBB6F(1)-Kit(W)/Kit(W-v) and WBB6F(1)-Mitf(mi-vga9)/Mitf(mi-vga9) mice.