Motomichi Sasaki

Localization of a novel v-erbB-related gene, c-erbB-2, on human chromosome 17 and its amplification in a gastric cancer cell line.

The c-erbB-2 gene is a v-erbB-related proto-oncogene which is distinct from the gene encoding the epidermal growth factor receptor. By using two independent methods, hybridization of both sorted chromosomes and metaphase spreads with cloned c-erbB-2 DNA, we mapped the c-erbB-2 locus on human chromosome 17 at q21, a specific breakpoint observed in a translocation associated with acute promyelocytic leukemia. Furthermore, we observed amplification and elevated expression of the c-erbB-2 gene in the MKN-7 gastric cancer cell line. These data suggest possible involvement of the c-erbB-2 gene in human cancer.

A phylogenetic study of bird karyotypes

Karyotypes were compared in 48 species, including 6 subspecies, of birds from 12 orders: Casuariiformes, Rheiformes, Sphenisciformes, Pelecaniformes, Ciconiiformes, Anseriformes, Phoenicopteriformes, Gruiformes, Galliformes, Columbiformes, Falconiformes and Strigiformes. — With the exception of the family Accipitridae, all the species studied are characterized by typical bird karyotypes with several pairs of macrochromosomes and a number of microchromosomes, though the boundary between the two is not necessarily sharp. The comparative study of complements revealed that a karyotype with 3 morphologically distinct pairs of chromosomes is frequently encountered in all orders except the Strigiformes. Those 3 pairs, submetacentric nos. 1 and 2, and a subtelocentric or telocentric no. 3, are not only morphologically alike but also have conspicuous homology revealed by the G-banding patterns. Furthermore, G-banding analysis provided evidence for the derivation of the owl karyotype from a typical bird karyotype.—The above cytogenetic features led to the assumption that the 3 pairs of marker chromosomes had been incorporated into an ancestral bird karyotype. It seems probable that those chromosomes have been transmitted without much structural changes from a common ancestor of birds and turtles, since the presence of the same marker chromosomes in the fresh water turtle Geoclemys reevesii is ascertained by G-banding patterns. — A profile of a primitive bird karyotype emerged through the present findings. Hence, it has become possible to elucidate mechanisms involved in certain structural changes of macrochromosomes observed in birds. It was concluded that a major role had been played by centric fission as well as fusion, translocation, and pericentric inversion.

Regional and temporal changes in the pattern of X-chromosome replication during the early post-implantation development of the female mouse.

We have made a detailed study of the X-chromosome replication pattern during the period when X-inactivation is occurring in the mouse embryo. Our observations show unequivocal regionalization of the embryo with respect to the temporal appearance, parental origin and DNA replication pattern of the allocyclic X-chromosome. The switch from isocyclic to allocyclic replication occurs in the embryonic ectoderm at approximately 6 days of development and is random with respect to parental origin of the X-chromosome. In the extra-embryonic tissues, however, the switch to allocyclic replication has apparently occurred prior to 5.3 days of development and almost exclusively involves the paternally-derived X-chromosome. Since these findings are consistent with results obtained in biochemical studies of X-chromosome activity in female embryos, we conclude that there is a close temporal relationship between the cytogenetic and biochemical manifestations of the X-inactivation process. In addition, we have observed a pattern of early paternal X-chromosome replication, transitory in some cases, that is unique to extra-embryonic tissues. These results suggest that there may be some differences in the mechanism by which X-inactivation occurs in the extra-embryonic tissues as compared with the embryonic ectoderm.

Location of Nucleolar Organizers in Animal and Plant Chromosomes by Means of an Improved N-Banding Technique

With an improved N-banding technique, the location of nucleolar organizing region was determined in 27 kinds of material including mammals, a marsupial, birds, amphibians, fishes, an insect and plants. In most cases the N-bands were clearly located on certain specific regions of chromosomes, such as the secondary constriction, satellite, centromere, telomere and heterochromatic segment, while in some species they were detected as very minute bodies distributed over many chromosomes. From the available cytological and biochemical data it was suggested that the N-bands represent certain structural non-histone proteins specifically linked to nucleolar organizers in various eukaryotic chromosomes.

Reversal of X-inactivation in female mouse somatic cells hybridized with murine teratocarcinoma stem cells in vitro

A series of near-diploid embryonal carcinoma-like hybrid cells were obtained from polyethylene glycol mediated cell fusion between murine embryonal carcinoma cells (PSA-6TG1 or OTF9-63) having one X chromosome and thymocytes or bone marrow cells from female mice carrying Cattanachs or Searles translocation. Prior to fusion with EC cells the somatic cells are presumed to contain only one active X chromosome. Following hybrid formation, the chronology of X chromosome replication and the expression of X-linked gene Pgk-1 indicated that all X chromosomes contributed by both parents were active in these hybrids. Experiments were performed to rule out the possibility that the hybrids were formed by fusion of EC cells with rare somatic cells in which both X chromosomes were active. Taken together the data indicate that within four days of fusion there is reactivation of the entire inactive X chromosome.

High Susceptibility to Hepatocellular Carcinoma Development in LEC Rats with Hereditary Hepatitis

A remarkably high incidence of hepatocellular carcinomas was observed in long‐surviving LEC rats with hereditary hepatitis. Among the 60 LEC rats examined between 12 and 28 months of age from F29, and F30, 55 (92%) developed putative preneoplastic and neoplastic lesions such as hyperplastic foci and nodules, and hepatocellular carcinomas. Of these, hepatocellular carcinomas were observed with a high frequency (46/55; 84%). All rats of advanced age that survived more than 18 months developed hepatocellular carcinomas. These results suggest that the development of liver tumors in LEC rats is an age‐associated phenomenon with serial hepatic alterations after the subsidence of acute hepatitis. The long‐surviving rats had no normal tissue and showed chronic hepatitis in nontumorous tissues of the liver. Cholangiofibrosis was also found in most rats with hepatic lesions. Metastasis of hepatocellular carcinomas was found in four rats. Histologically, the hepatocellular carcinomas were of a well‐differentiated type with a typical trabecular structure. Thus, LEC rats seem to be a promising animal model for studying the pathogenesis of hepatitis and hepatocellular carcinoma.

AN IMPROVED SILVER STAINING TECHNIQUE FOR NUCLEOLUS ORGANIZER REGIONS BY USING NYLON CLOTH

SummaryA simple and reproducible silver-staining technique for nucleolus organizer regions (NORs) was developed, use being made of nylon cloth as a coverslip for even impregnation of the silver solution. Ag-NORs were clearly and selectively visualized in human and mouse chromosomes, without equivocal staining of centrometric heterochromatin and background silver grains.

Cytologic evidence for preferential inactivation of the paternally derived x chromosome in xx mouse blastocysts.

A total of 941 mouse blastocysts obtained from two types of crosses in which one of parents carried Cattanachs X/autosome translocation was studied cytogenetically by quinacrine mustard fluorescence. The rearranged X (Xt) and the normal X (Xh) were distinguished by size. Karyotype analysis was successful in 721 embryos, of which 205 were heterozygous for Cattanachs translocation. A single heterochromatic and brightly fluorescent X chromosome was identified in 154 metaphase spreads from 89 blastocysts consisting of 32--96 cells. The paternally derived X chromosome (Xp) was heterochromatic in 87% and 88% of the informative cells from the crosses XnXn x XtY and XtXn x XnY, respectively. This preferential choice of Xp at the blastocyst stage might have an important bearing upon the preponderance of cells with an inactive Xp in the chorion and yolk-sac splanchnopleure.

Chromosome studies in four species of Ratitae (Aves)

Chromosomes were studied in female specimens of the ostrich, Struthio camelus L., cassowary, Casuarius casuarius (L.), emu, Dromiceius novaehollandiae (Lath.) and rhea, Rhea americana L. by means of blood and feather pulp culture techniques. Male karyotypes were also studied in the emu and rhea. The diploid chromosome number was most likely 80 in the ostrich and rhea and 82 in the emu, while the exact number could not be determined in the cassowary. Karyotypes of the 4 species were strikingly similar and apparently interchangeable with one another with slight modifications of the centromeric position in one or two pairs of macrochromosomes. No heteromorphic macrochromosomal pair was found either in female specimens or in male ones of the ratite species so far examined, except for a female rhea. This specimen was found to possess an acrocentric chromosome which was evidently a member of nos. 4–6, but considerably smaller than any other chromosome of the group. 3H-thymidine autoradiography provided no more information than the straightforward morphological analysis with regard to the differentiation of the sex-chromosomes.

A summary of cytogenetic studies on 534 cases of chronic myelocytic leukemia in Japan

Cytogenetic and clinical data on 534 patients with chronic myelocytic leukemia (CML) were collected from 10 institutions in Japan. The results of the analysis of the data were in substantial accord with those of the First International Workshop on Chromosomes in Leukemia and other published data, but certain differences were noted in the frequency of Philadelphia chromosome (Ph1)-negative cases, unusual and complex Ph1 translocations, and additional chromosome changes. Some of the findings are discussed with respect to the origin of unusual and complex Ph1 translocations, the relationship between chromosome abnormalities and survival, and geographic differences in chromosome abnormalities.