The mutant gene "wal" is active in cells of mouse hair follicles

In order to determine the place of action of the mutant gene waved alopecia (wal), we have obtained chimeric wal/wal c/c Gpi-1aa<-->+/+ C/C Gpi-1bb animals by aggregation of eight-cellular embryos of BALB/c-wal/wal mice and CBA (+/+) mice. The presence or absence of the chimeric structure was determined from the mosaic nature of fur color and hair structure, as well as on the basis of the presence of electrophoretically distinct variants of glucosephosphate isomerase in blood. Chimeras had alternating transverse patches of different lengths and widths consisting of curly (genotype wal/wal) or straight (genotype +/+) hairs. The percentage of cells with wal/wal mutant genotype in chimeras established on the basis of glucosephosphate isomerase isozymes varied from 10 to 80%. A higher percentage of the parental wal/wal component in chimeras correlated with the number of patches having wavy hairs. Analysis of the fur pattern represented by the alternation of transverse patches of wavy or straight hairs in chimeric wal/wal (+/+ mice has shown that mutant gene wal acts in ectodermal cells of hair follicles.     

Mutant gene expression in murine aggregation chimeras. 5. The ocular retardation and fidget genes

Analysis of ocular retardation (or) and fidget (fi) genes expression in 18 day old embryos, 10 and 20 day old or/or C/C----+/+ c/c and fi/fi or/or C/C----+/+ +/+ c/c mice has shown that genes or and fi are active in developing retina and suppress cell proliferation. Structural defects of retina and decrease in the eye size in the chimaeras, compared to the normal embryos, were observed already in the presence of 13-16% of mutant cells. As the fraction of mutant cells increased, the degree of eye disturbances increased as well. In the fi/fi or/or----+/+ +/+ chimaeras structural defects of retina and decrease in the eye size are more pronounced than in the or/or----+/+ chimaeras, due to the synergetical effect of both mutant genes in the fi/fi or/or cell clones. In the ontogenesis of the or/or----+/+ chimaeras the development of the retinal photoreceptor layer is normalized due to the substitution of mutant cells for actively proliferating normal cells. No metabolic cooperation between the mutant and normal cells was observed in the developing retina of chimaeras.     

Expression of mutant genes in mouse aggregation chimeras. 4. The aphakia gene

Analysis of aphakia (ak) gene expression in ak/ak C/C in equilibrium +/+ c/c experimental chimaeras has shown that the ak gene acts in the lens rudiment cells blocking it differentiation. In the lens of 12 day old ak/ak C/C in equilibrium +/+ c/c chimaeric embryos undifferentiated ak/ak cells were present among the normally differentiating fibres. In 14 and 18 day old chimaeric embryos and 20 day old chimaeric mice ak/ak cells are located under the lens epithelium and the capsule of posterior lens half. In the locations of ak/ak cells on the posterior lens surface capsule breaks resulted in the extrusion of lens material into the secondary eye cavity. In all studied chimaeric embryos the lens structure is more similar to that in the normal embryos, than in ak/ak embryos. This suggests that in the developing chimaeric lens ak/ak cells are sorted out as the development proceeds. The proliferation rate of +/+ cells appears to be higher than that of ak/ak cells.     

Interaction of the mutant aphakia, fidget and ocular retardation genes in mice

The phenogenetic analysis of the effects of aphakia (ak) gene and its interaction with the ocular retardation (or) and fidget (fi) genes suggests that the ak gene acts in the lens cells with the result of arresting lens fibre differentiation. In mice homozygous for ak, the lens failure leads to secondary retina defects, in particular, to formation of retinal folds. In ak/ak or/or mice, the lens and retina morphogenesis stops at the optic cup stage, the eye is strongly reduced in size and more affected, compared to the corresponding single homozygotes. Unlike ak/ak or/or, in the ak/ak fi/fi mice the eyes are more regular in shape than those in the ak/ak +/+ condition. The fi gene inhibition of the retina anlage growth leads to some improvement of the eye development in double ak/ak fi/fi homozygotes, due to the absence of extensive retina folding.     

Collagenase-3 expression in periapical lesions: an immunohistochemical study

Collagenase-3 (matrix metalloproteinase-13) is a metalloproteinase (MMP) that is associated with bone lesions and exhibits variable expression patterns in odontogenic cysts; it may play a role in regulating focal proliferation and maturation of jaw cyst epithelium. We studied the localization, staining intensity and distribution of collagenase-3 in 13 periapical granulomas with epithelium, 16 periapical granulomas without epithelium and 10 radicular cysts using archived formalin fixed, paraffin embedded tissues. A monoclonal antibody against human collagenase-3 was used to evaluate its expression. Immunohistochemical staining intensities of collagenase-3 in all periapical lesions were (?), 4 (10%); (+), 1 (3%); (++), 22 (56%) and (+++), 12 (31%); differences were not statistically significant. Immunohistochemical distribution of collagenase-3 in epithelial cells was (?), 17 (44%); (+), 17 (44%); (++), 5 (13%); in fibroblasts it was (?), 8 (20%); (+), 23 (59%); (++), 8 (21%); in plasma cells it was (?), 7 (18%); (+), 22 (56%); (++), 10 (26%); in macrophages it was (?), 7 (18%); (+), and 15 (38%); and (++), 17 (44%). Statistically significant differences were found in epithelial cells (p = 0.00) and fibroblasts (p = 0.02), whereas differences were not statistically significant for plasma cells and macrophages. Collagenase-3 may play a role in the conversion of a periapical granuloma with epithelium to radicular cyst. MMP's influence not only epithelial rest cell migration, but also invasion of various stromal cells into granulomatous tissue.     

Expression of mutant eyeless genes in the mouse embryo retina

The aim of the present study was to determine the cellular site of eyeless-I (ey-I) and eyeless-2 (ey-2) gene action, causing anophthalmia or microphthalmia. Eye primordia from 10-day-old embryos of ZRDCT-AN and CC57BR (control) mice were cultured in vitro for 3 or 6 days. In 59 out of 77 cultured mutant eye primordia neural retina was disturbed. In 9 mutant eye primordia the disturbed neural retina was 4-6 times thinner than in the control. However, lens differentiation was similar to that in the control, epithelial and fibrous components were observed. Thus, mutant genes eyeless inhibit the growth of primordial retina, causing secondary developmental defects of the lens and other eye structures.     

Analysis of silver binding nucleolar organizer regions in exfoliative cytology smears of potentially malignant and malignant oral lesions

Nucleolar organizer regions are nucleolar components that contain proteins that are stained selectively by silver methods; they can be identified as black dots throughout the nucleolus and are known as silver binding nucleolar organizer regions (AgNOR). The number of AgNOR is related to the cell cycle and the proliferative activity of the cells. We investigated AgNOR using exfoliative cytology smears of potentially malignant oral lesions. Eighty individuals were divided into four equal groups: healthy controls, oral leukoplakia, oral submucous fibrosis and oral squamous cell carcinoma. The mean number of AgNOR in each study group gradually increased from control to oral leukoplakia to oral submucous fibrosis to oral squamous cell carcinoma. The proliferative index was increased in the oral premalignant and malignant patients compared to normal subjects. The mean AgNOR size gradually increased from control to oral leukoplakia to oral submucous fibrosis to oral squamous cell carcinoma. Spherical shaped AgNOR were most common in controls, whereas large, clustered and kidney shapes were most common in oral squamous cell carcinoma. Multiparameter analysis of AgNOR in oral exfoliative smears is a simple, sensitive and cost-effective method for differentiating premalignant from malignant lesions and can be used in conjunction with routine cytomorphological evaluation.     

Effects of the mutant gene wellhaarig in chimeric mice

The mutant gene wellhaaring (we) confers the waved coat in mice, which is most pronounced in homozygotes at 10 to 21 days of postnatal development. Abnormal hair growth and structure in the we/we mutant mice results from defective cell differentiation in the inner root sheath of a hair follicle. To localize the site of the we gene action, we obtained ten chimeric mice by aggregation of the early C57BL/6-2we/we and BALB/c embryos. The chimera coat was waved, shaggy, or almost normal depending on the percentage of the mutant component. In the we/we +/+ chimeric animals of the first generation (G1) aged 21 days, both mutant and normal hair phenotypes were observed, which was especially discernible in zigzag hair. Note that none of the chimeras exhibited the alternating patterns of transversely oriented stripes or patches of either mutant or normal hair; i.e., they had a mixed parental hair phenotype. We also did not observe the animals with an intermediate phenotype, which suggests a discontinuous hair formation in chimeras according to the "all or nothing" principle. The data obtained indicate that the dermal papilla cells of a hair follicle are the sites for the we gene action. During the embryonic development, dermal cells are strongly mixed, which accounts for the lack of the clear-cut transverse stripes of either mutant or normal hair. The mutant gene we is probably responsible for a disrupted induction signal from the dermal papilla towards ectodermal cells of a hair follicle.     

Genomic imprinting in mammals

A review of the data on the mechanisms and effects of genomic imprinting, an epigenetic phenomenon regulating the development in placentate mammals, is presented. In contrast to the majority of gene loci with biallelic expression, the expression of imprinted loci is monoallelic. In humans and mice, more than 300 imprinted loci have been identified, in which maternal or paternal alleles may either be expressed or be found in a repressed state during ontogeny. Imprinting is established during gametogenesis, and the repression of an allele of the imprinted locus is determined by methylation of the key regulatory element of this allele. Both the maternal and paternal chromosome sets are required for normal development in mammals. This is why parthenogenesis and androgenesis in these animals are impossible in nature. As a result of differential gene expression of many imprinted loci, the balance of gene activity is established, which is necessary for normal proliferation and differentiation of various cell clones in embryogenesis. Many human developmental abnormalities and syndromes are determined by defective genomic imprinting. In particular, the loss of imprints, which is followed by the occurrence of biallelic expression of some imprinted loci, may cause malignant tumors.