Detection of nucleolus organizer regions in chromosomes of human,chimpanzee, gorilla,orangutan and gibbon

Nucleolus organizer regions were detected by the Ag-AS silver method in fixed metaphase chromosomes from human and primates. In the human, silver was deposited in the secondary constriction of a maximum of five pairs of acrocentric chromosomes: 13, 14, 15, 21 and 22. The chimpanzee also had five pairs of acrocentric chromosomes stained, corresponding to human numbers 13, 14, 18, 21 and 22. A gibbon had a single pair of chromosomes with a secondary constriction, which corresponded to the nucleolus organizer region. In each case the Ag-AS method detected the sites which have been shown by in situ hybridization to contain the ribosomal RNA genes. An orangutan had eight pairs of acrocentric chromosomes stained with Ag-AS, probably corresponding to human numbers 13, 14, 15, 18, 21 and 22, plus two others. Two gorillas had silver stain over two pairs of small acrocentric chromosomes and at the telomere of one chromosome 1. The larger gorilla acrocentric chromosomes had no silver stain although they all had secondary constrictions and entered into satellite associations.     

Restriction endonuclease resistant chromatin in human chromosomes

Summary The recent addition of restriction endonucleases in obtaining selective bands in the human genome has added a new dimension to molecular genetics. However, a considerable discrepancy exists in banding patterns produced by AluI in chromosomes 19 and 20, by MboI in chromosomes 4, 5, 8, 21 and 22 and by RsaI in chromosomes 12, 21 and 22. The principal causes of these differences are highlighted.     

The banding pattern produced by restriction endonucleases in mouse chromosomes

The metaphase chromosomes of mouse have been treated with the restriction endonucleases Alu I, Mbo I, Hae III and Eco RII and subsequently stained with the DNA-specific dye Ethidium bromide. A striking correspondence between previous biochemical findings and our cytological results has been noticed with Alu I, Mbo I and Hae III, which were capable of digesting all but the DNA localized at the centric constitutive heterochromatic areas. Digestion with Eco RII, on the contrary, resulted in cytological data which apparently did not fit in with the biochemical results previously obtained by digesting the DNA of this species with the same enzyme. It is postulated that factors such as chromatin organization, in addition to DNA base sequence, can determine the results we report.     

The mechanism and pattern of banding induced by restriction endonucleases in human chromosomes

The mechanism of chromosome banding induced by restriction endonucleases was analyzed by measuring the amount of radioactivity extracted from [¹⁴C]thymidine-labeled chromosomes digested first with restriction enzymes and subsequently with proteinase K and DNase I. Restriction enzymes with a high frequency of recognition sites in the DNA produced a large number of short DNA fragments, which were extracted from chromosomes during incubation with the enzyme. This loss of DNA resulted in decreased chromosomal staining, which did not occur in regions resistant to restriction enzyme digestion and thus led to banding. Subsequent digestion of chromosomes with proteinase K produced a further loss of DNA, which probably corresponded to long fragments retained in the chromosome by the proteins of fixed chromatin. Restriction enzymes induce chromatin digestion and banding in G1 and metaphase chromosomes, and they induce digestion and the appearance of chromocenters in interphase nuclei. This suggests that the spatial organization and folding of the chromatin fibril plays little or no role in the mechanism of chromosome banding.It was confirmed that the pattern of chromosome banding induced by AluI, MboI, HaeIII, DdeI, RsaI, and HinfI is characteristic for each endonuclease. Moreover, several restriction banding polymorphisms that were not found by conventional C-banding were detected, indicating that there may be a range of variability in the frequency and distribution of restriction sites in homologous chromosome regions.     

Panels of somatic cell hybrids specific for chimpanzee, gorilla, orangutan, and baboon

The generation of panels of somatic cell hybrids specific for chimpanzee, gorilla, orangutan, and olive baboon is reported. The chromosome content of each hybrid clone was characterized using reverse painting on human normal metaphases and by the use of appropriate sequence tag sites (STSs), one for each chromosome arm. These resources can be advantageously exploited in the characterization of chromosome architecture of different primate species, with special reference to the discrimination of inter- and intra-chromosomal arrangement of segmental duplications.     

A human-derived probe,p82H,hybridizes to the centromeres of gorilla,chimpanzee, and orangutan

A human-derived centromeric sequence, p82H, hybridizes to DNA from gorilla, chimpanzee, pygmy chimpanzee, and orangutan. On DNA blots, multimeric ladders based on 170 or 340 bp repeat units are seen. In metaphase chromosome preparations from these species, p82H hybridizes to the centromeric region of every chromosome. p82H forms less stable hybrids with DNA from the lion-tailed macaque and does not hybridize to DNA or chromosomes of the owl monkey or the mouse.     

Confirmation of the specific status ofHapalemur aureus (Primates,Strepsirhini) by restriction genomic DNA banding patterns

This study confirms, on the basis of our molecular biology results and in accordance with cytogenetic, morphological and ethological data, the specific status ofHapalemur aureus. Furthermore, it appears clearly thatHapalemr simus began its differentiation fromHapalemur griseus griseus andHapalemur aureus (wich have a common branch) shortly after the separation ofLemur catta from the phylogenetic tree of theL. catta/ Hapalemur group.     

5-Methylcytosine in heterochromatic regions of chromosomes: chimpanzee and gorilla compared to the human

Fixed metaphase chromosomes of gorilla and chimpanzee were UV-irradiated to produce regions of single-stranded DNA and then treated with antibodies specific for the minor DNA base 5-methylcytosine (5 MeC). An indirect immunofluorescence technique was used to visualize sites of antibody binding. In the gorilla six pairs of autosomes contained major fluorescent regions, indicating localized regions of highly methylated DNA. These corresponded, with the exception of chromosome 19, to the major regions of constitutive heterochromatin as seen by C-banding. The Y chromosome also contained a highly fluorescent region which was located just proximal to the intense Q-band region. In the chimpanzee no comparable concentrations of highly methylated DNA were seen. Smaller regions of intense 5 MeC binding were present on perhaps six chimpanzee chromosomes, including the Y. Five of these corresponded to chromosomes which were highly methylated in the gorilla.--There is diversity among the human, gorilla and chimpanzee in both the size and location of concentrations of 5 MeC, supporting the idea that satellite DNA evolves more rapidly than DNA in the remainder of the chromosome.     

Cytological identification of the chromosomes involved in Nishimura's rice translocation lines

During the past three decades, Nishimura's reciprocal translocation lines of rice have been used in rice cytogenetics to locate genes on chromosomes, to number extra chromosomes of trisomic series and to associate individual linkage groups with specific chromosomes. In this report, we present our identification of the chromosomes involved in 11 of Nishimura's translocation lines using both meiotic pachytene and mitotic prometaphase chromosome analysis. In addition, the numbering of the 12 linkage groups suggested by Nagao and Takahashi, and modified later by many workers, has been revised to agree with the numbering of the identified chromosomes.     

The banding pattern of the salivary gland chromosomes of Drosophila hydei

The banding pattern of the salivary gland chromosomes of D. hydei was investigated in the electron microscope. We compared the banding pattern of squashed chromosomes with non-squashed preparations and observed that the fixation and squash procedure we used does not introduce artificial changes in the banding pattern of the chromosome. An electron microscopic map was made of the banding pattern of the distal half of the second salivary gland chromosome. On the basis of the number of bands in this part of the second chromosome we calculated a total of about 3700 bands for the whole set of polytene chromosomes of D. hydei. Our data indicate a similar number of bands in the salivary gland chromosomes of evolutionary remote Drosophila species like D. hydei and D. melanogaster.     

The giemsa banding pattern of canine chromosomes, using a cell synchronization technique

Cytogenetic investigations of the domestic dog, Canis familiaris, were performed on the Doberman pinscher and two Boxer dogs. Conventional homogeneously stained and G-banded metaphases from peripheral blood lymphocyte cultures synchronized with amethopterin and bromodeoxyuridine were studied. These procedures permitted the unequivocal identification of all canine chromosomes. A canine chromosome idiogram was constructed on the basis of the G-banding pattern at the haploid 327-band resolution level. The secondary constrictions and tapering of the telomeric regions characteristic of several canine chromosomes are described. Q-, C-, and NOR-banding were also performed and the salient features are described. This karyotype should enhance the value of the canine species in cytogenetic investigations.     

Sulfonation of polyvinylidene difluoride resin and its application in extraction of restriction enzymes from DNA digestion solutions

Sulfonation of polyvinylidene difluoride (PVDF) resin was achieved by incubation of the resin with sulfuric acid at a moderately high temperature. The sulfonated PVDF (SPVDF) resin was studied for its ability to extract restriction enzymes from DNA digestion solutions. The SPVDF resin was effective in adsorbing restriction enzymes such as EcoRI and BamHI and the extraction procedure was easy and simple to perform. The adsorption depended upon the amount of the resin added. We found that 1 mg of the SPVDF resin could completely remove all restriction enzyme activity routinely used in DNA digestion within 2 min after its addition. Treatment of a digestion solution with the SPVDF resin did not change the reaction solution and the same digestion buffer could be used for another digestion of the same DNA with other enzymes. We also found that, in comparison with normal PVDF, the SPVDF resin adsorbed less DNA, resulting in less loss of DNA in the extraction step. The potential application of the SPVDF resin in other procedures of molecular cloning and enzyme purification is discussed.     

大赖草及近缘种染色体C—分带的研究

对Ｔｈｉｎｏｐｙｒｕｍｎｂｅｓａｒａｂｉｃｕｍ（Ｓａｖｕｌ．＆Ｒａｙｓ）Ａ．Ｌｖｅ、新麦草（Ｐｓａｔｈｙｒｏｓｔａｃｈｙｓｊｕｎｃｅａ（Ｆｉｓｃｈ．）Ｎｅｖｓｋｉ）和大赖草（Ｌｅｙｍｕｓｒａｃｅｍｏｓｕｓ（Ｌａｍ．）Ｔｚｖｅｌ．）染色体Ｃ分带的核型进行了比较研究。Ｔｈ．ｂｅｓａｒａｂｉｃｕｍ和新麦草的染色体在Ｃ分带带型上有明显的差异,显示了物种的特异性。３个物种的核型表明,Ｃ带带纹主要分布在染色体的末端,大部分染色体不显着丝粒带和中间带。在大赖草染色体上的末端带很明显。一些大赖草的染色体具有与Ｔｈ．ｂｅｓａｒａｂｉｃｕｍ和新麦草某些染色体相似的Ｃ带带型。对大赖草染色体组与Ｔｈ．ｂｅｓａｒａｂｉｃｕｍ和新麦草染色体组的关系进行了讨论。     

The chromosomes of the Didelphidae (Marsupialia) and their evolutionary significance

One hundred and seventy-seven specimens of American didelphids, representing 9 genera and 22 species have been studied for their chromosomal constitution. Didelphids are very conservative in chromosomal complements. All of the studied species can be sorted into one of three kinds of karyotypes: 2 n = 14 (three species of Didelphis, one of Lutreolina, two of Philander, and one of Chironectes) , 2 n = 14 (eight species of Marmosa, one of Metachirus, three of Caluromys, and one of Dromiciops), and 2 n = 18 (three species of Monodelphis). These karyotypes are stable, showing only minor variations within each basic pattern. It is concluded that chromosomals evolution in the Didelphidae proceededs from low numbers to higher numbers by a process of centromeric fissioning complemented by some pericentric inversions and/or translocations. The pattern of karyotypic stability is consistent with bradytely at the organismic level of evolution. This is explained by a low rate of regulatory genetic evolution promoted by epistatic selection favouring the retention of chromosomal arrangements highly advantageous for overall adaptation.     

Histological study on the chronology of the developing dentition in gorilla and orangutan

The single previous study on tooth development in great apes (Dean and Wood: Folia Primatol. (Basel) 36:111–127, 1981) is of limited value because it is based on cross-sectional radiographic data. This study considers problems in defining stages of tooth development in radiographs of developing ape dentitions and provides data on tooth chronology in Pongo pygmaeus and Gorilla gorilla by using histological methods of analysis. Crown formation times were estimated in individual teeth, and an overall chronology of dental development was found by registering teeth forming at the same time by using incremental growth lines. The earlier radiographic study correctly identified the molar and second premolar chronology and sequence in great apes, but significantly underestimated crown formation times in incisors, first premolars, and canine teeth in particular. Ape anterior tooth crowns take longer to form than the equivalent human teeth, but the overall dental developmental period in great apes is substantially shorter than in humans. Gorilla root extension rates appear to be fast, up to approximately 13 μm/day. This rapid root growth, associated with early tooth eruption, appears to be the developmental basis for the observed differences in timing between developing dentitions in great apes and humans.