Banding pattern analysis of polymorphic karyotypes in the black rat by a new differential staining technique

Polymorphic karyotypes of black rats (Rattus rattus) collected in Japan, Australia and India were analysed by a new differential staining technique by which banding patterns in the metaphase chromosomes are revealed. The technique consists in two steps: immersion of slides in a mixture of 2 x SSC and 0.1% (w/v) SDS (sodium dodecyl sulfate) for a few seconds at room temperature, and staining in Giemsa. By this treatment characteristic banding patterns were obtained in each chromosome pair. From the banding pattern analysis, subtelocentric pairs No. 1 and 9, which are polymorphic in respect to the acrocentrics and the subtelocentrics, were proven to have originated by pericentric inversion in the acrocentrics. The origin of two large metacentrics observed in Australian and Indian black rats was confirmed to have been developed by Robertsonian fusion of the acrocentrics No. 4 and 7 and No. 11 and 12 present in the Asian type black rat.Contribution No. 873 from the National Institute of Genetics, Japan. Supported by a grant-in-aid from the Ministry of Education of Japan (Nos. 92159 and 92332).     

Banding patterns of Chinese hamster chromosomes revealed by new techniques

Two kinds of techniques were newly developed to reveal banding patterns of the Chinese hamster chromosomes. Both techniques were essentially the same as those used for the extraction of proteins, and one of them could induce bands in chromosome arms in only a few seconds. Banding patterns produced by these techniques appeared to be identical to those induced by the methods reported by previous workers, requiring post-fixation incubation of slides in a warm saline. —The banding pattern was typical for each chromosome pair, permitting unequivocal identification of several pairs which were hardly distinguished by the conventional staining procedures. It was confirmed that these patterns had been well preserved in the chromosomes of a cultured cell line.Contribution No. 870 from the National Institute of Genetics, Japan. This work was supported in part by a grant (No. 9001) from the Ministry of Education in Japan.     

茅舍血厉螨核型及染色体的C带、G带的研究

本文首次报道了一种革螨——茅舍血厉螨核型及染色体C带、G带的研究。用剖腹取卵法、玻璃纸压片、Giemsa染色,经分析茅舍血厉螨的核型,单倍体n=7,二倍体2n=14。 用氢氧化钡—吉姆萨技术显示茅舍血厉螨染色体C带。在第1、2、4、5染色体上出现恒定的C带部分,第3、6、7染色体上出现不恒定的C带部分。根据C带带型,茅舍血厉螨着丝点的位置可分为近中区域(sm),近端区域(St),末端区域(t)和末端点(T)四类。 G带分析用胰蛋白酶—吉姆萨技术显示。 本文对茅舍血厉螨的核型、革螨染色体研究中螨卵的收集方法和染液的改进、C带带型与着丝粒位置的确定和G带显带问题进行了讨论。     

The karyotype of the mouse

A denaturating and renaturating technique, applied to mouse chromosomes, makes visible characteristic banding patterns by which all elements of the karyotype can be individually distinguished. The Y chromosome as a whole appears darkly stained. The X chromosome comprises 6.33% of the homogametic haploid set. The banding pattern of the chromosomes is compared with that obtained by aid of the quinacrine dihydrochloride fluorescence technique. After its use a banding pattern results which is similar to, but less distinct than, that found after the renaturation procedure.     

Giemsa banding,quinacrine fluorescence and DNA-replication in chromosomes of cattle (Bos taurus)

Almost all the 30 chromosome pairs of cattle can be identified by their banding patterns made be visible by a Giemsa staining technique described previously. The banding pattern of the X chromosome shows striking similarities with the banding pattern of the human X chromosome. — The centromeric region of the acrocentric autosomes contains a highly condensed DNA. This DNA is removed by the Giemsa staining procedure as can be shown by interference microscopic studies. If the chromosomes are stained with quinacrine dihydrochloride these centromeric regions are only slightly fluorescent. — Autoradiographic studies with ³H-thymidine show that the DNA at the centromeric regions starts and finishes its replication later than in the other parts of the chromosomes.     

大熊猫与黑熊显带染色体的比较研究

以体外培养的大熊猫（Ａｉｌｕｒｏｐｏｄａｍｅｌａｎｏｌｅｕｃａ）与黑熊（Ｓｅｌｅｎａｒｃｔｏｓｔｈｉｂｅｔａｎｕｓ）外周血淋巴细胞为实验材料,应用ＢｒｄＵ复制带显示技术,研究了大熊猫和黑熊染色体晚复制带带型。通过对大熊猫与黑熊显带染色体带型的比较,发现黑熊部分具端着丝粒的染色体与大熊猫部分具中,亚中,或亚端着丝粒的染色体的整个短臂或整个长臂有明显的带型相似性,在黑熊具中,亚中着丝粒染色体中,仅３３     

Banding of human chromosomes with basic fuchsin

Summary In this paper a technique is described for the banding of human metaphase chromosomes with basic fuchsin. The main characteristics of the G-banding pattern obtained with this cationic triphenylmethane dye are:the secondary constriction regions of chromosomes Nos. 1 and 16 are strongly stained, especially in the latter one;the heterochromatic area of chromosome No. 9, usually negative with most other G-banding techniques, is clearly visible as an intensely stained band adjacent to the centromere;the chromosomal outline is often very distinct, facilitating the study of the telomeres; a number of chromosomal regions with bright Q fluorescence such as the polymorphic regions of the chromosomes Nos. 3, 4, and Y also stain strongly with basic fuchsin.The basic fuchsin technique combines therefore properties of G-, C-, and Q-banding methods and seems very suitable for use in e.g., family and linkage studies.Several triphenylmethanes closely related to basic fuchsin produce similar banding patterns. The band-producing ability is, however, diminished in those dyes which contain methylated amino groups. If the methyl groups are attached to the carbon atoms at the 3-positions in the phenyl rings, band formation seems unaffected.The way in which basic fuchsin and chromatin may interact as well as the possible mechanism(s) of band formation with this dye are discussed.     

Similarity of giemsa banding patterns of chromosomes in several species of the Genus Rattus

Giemsa banding patterns of chromosomes in seven Rattus species were compared. Four species (R. rattus tanezumi, R. norvegicus, R. exulans and R. muelleri) had all 2n=42 and their karyotypes and banding patterns were similar, although slight differences were observed. Another subspecies (R. rattus rattus) and two other species (R. fuscipes and R. conatus) had fewer chromosomes than the above species by having large biarmed chromosomes developed probably by Robertsonian fusion. The origin of the arms of biarmed chromosomes was recognized by their characteristic banding patterns. The remaining species, R. sabanus, had a karyotype markedly different from the other species by having two small metacentrics although in the others their number was 7. Banding patterns of the chromosomes in this species, however, were also very similar to those of the other, and therefore the 7 small metacentrics seemed to have originated by pericentric inversion of small acrocentrics.Contribution No. 912 from the National Institute of Genetics, Japan. Supported by a grant-in-aid from the Ministry of Education of Japan, Nos. 90183, 90375 and 744002.     

Structural organization of chromosomes of the Indian muntjac (Muntiacus muntjak)

The identification, morphology, and banding pattern of the chromosomes of the Indian muntjac (Muntiacus muntjak) are described. A diagrammatic representation of the banding pattern as revealed by various techniques is presented following the nomenclature suggested by Paris Conference (1971) for human chromosomes. The Y2 chromosome and the neck of the X chromosome are late replicating based on observations made with the use of a bromodeoxuridine plus Giemsa technique. Most of the G-bands are early replicating, contrary to earlier findings based on autoradiography.     

An analysis of the technical variables in the production of C bands

Numerous combinations, concentrations, pH's and durations of HCl, NaOH and SSC treatments were tested for the purpose of developing an improved C banding technique for human metaphase chromosomes. Methods of slide preparation, as they affect C banding were also evaluated. — HCl and SSC treatment used separately, for all times and concentrations tested, gave no C banding. All treatment sequences which included an NaOH exposure gave at least some C banding, but also gave considerable swelling and distortion. Surprisingly, the best results were obtained from heat-dried preparations exposed to 0.2 N HCl at 25° C for 15 minutes, no NaOH and subsequently incubated in 2xSSC, pH=7.0 at 62–65° C for 18–24 hours. This technique is now being used routinely, following a G banding technique for homologue identification, to monitor C band variation in human chromosomes. — The pH of the 2xSSC incubation solution was found to be important. Slides treated as above with HCl, but with 2xSSC, pH=6.0 gave only G banding; HCl and 2xSSC, pH=8.0 gave C banding, but considerable chromosome swelling and poor uptake of stain. — Air- or ignition-dried preparations, with the HCl and 2xSSC treatment appeared undertreated and gave a mixture of G and C banding. A brief (30 second) exposure to 0.07 N NaOH between the HCl and 2xSSC steps is recommended. These results are in support of DNA-protein interaction and/or loss rather than denaturation-renaturation as a likely mechanism for C band production.     

The Giemsa banding pattern of the canine karyotype.

The canine metaphase karyotype consists of 78 chromosomes. All autosomes exhibit telocentric or acrocentric configurations gradually diminishing in size. These features make identification of homologous pairs by conventional analysis difficult. Chromosome preparations were derived from short-term cultures of peripheral blood lymphocytes obtained from clinically normal dogs representing at least four breeds. Most components of the canine karyotype can be distinguished readily. No significant G-banding pattern variations were detected in the individuals screened. An idiogrammatic interpretation of the banding pattern is presented. Apart from bands, other characteristic morphologic features were found which aid in identification. The G-banding pattern of the canine metacentric X is quite similar to that of the banded human X. The canine Y is a minute metacentric having two positive bands.     

家蚕染色体复制区带的初步显示

该文采用家蚕Bomoyx mori活体注射BrdU结合FPG(fluorochrome photolyusis Giem-sa)显带方法,以生殖腺为材料,成功显示出家蚕有丝分裂中期染色体复制带。由于处于S-期的细胞有早有晚,且同一细胞DNA各片段的复制亦有先后,因此BrdU掺入DNA合成的时间也有所不同,从而可产生出早、中、晚复制带型。BrdU掺入时间早,则会在家蚕部分染色体上出现大面积浅染带纹的早复制带。每一染色体皆有其独特的带纹特征,据此可初步将它与其它染色体相互区分;随着BrdU掺入时间的推后,染色体上会出现深浅交替、丰富的带纹,即中复制带型;至S-期DNA合成晚期掺入BrdU,最终染色体出现以深染带纹为主,浅染带纹仅出现于少数染色体的中部、近中部或端部的晚复制带。     

The pattern of restriction enzyme-induced banding in the chromosomes of chimpanzee,gorilla, and orangutan and its evolutionary significance

Summary The pattern of banding induced by five restriction enzymes in the chromosome complement of chimpanzee, gorilla, and orangutan is described and compared with that of humans. The G banding pattern induced by Hae III was the only feature common to the four species. Although hominid species show almost complete chromosomal homology, the restriction enzyme C banding pattern differed among the species studied. Hinf I did not induce banding in chimpanzee chromosomes, and Rsa I did not elicit banding in chimpanzee and orangutan chromosomes. Equivalent amounts of similar satellite DNA fractions located in homologous chromosomes from different species or in nonhomologous chromosomes from the same species showed different banding patterns with identical restriction enzymes. The great variability in frequency of restriction sites observed between homologous chromosome regions may have resulted from the divergence of primordial sequences changing the frequency of restriction sites for each species and for each chromosomal pair. A total of 30 patterns of banding were found informative for analysis of the hominid geneaalogical tree. Using the principle of maximum parsimony, our data support a branching order in which the chimpanzee is more closely related to the gorilla than to the human.     

Standard karyotype of sea trout (Salmo trutta morpha trutta) based on replication banding patterns

Replication banding patterns have been obtained from in vivo treatment of Salmo trutta morpha trutta chromosomes using a modification of the 5-BrdU technique and in kidney cultures using the fluorochrome photolysis Giemsa (FPG) staining method. Each chromosome pair was identified in the karyotype based on the banding pattern, chromosome size, and centromere position. The standard karyotype of sea trout has been proposed. Similarities of replication, and restriction enzyme banding patterns of chromosomes 8 and 9, and of chromosomes 11 and 12, are discussed. Fluorescence in situ hybridization was used to determine the location of telomeric sequences.     

Restriction endonuclease banding of rainbow trout chromosomes

Rainbow trout chromosomes were treated with nine restriction endonucleases, stained with Giemsa, and examined for banding patterns. The enzymes AluI, MboI, HaeIII, HinfI (recognizing four base sequences), and PvuII (recognizing a six base sequence) revealed banding patterns similar to the C-bands produced by treatment with barium hydroxide. The PvuII recognition sequence contains an internal sequence of 4 bp identical to the recognition sequence of AluI. Both enzymes produced centromeric and telomeric banding patterns but the interstitial regions stained less intensely after AluI treatment. After digestion with AluI, silver grains were distributed on chromosomes labeled with [³H]thymidine in a pattern like that seen after AluI-digested chromosomes are stained with Giemsa. Similarly, acridine orange (a dye specific for DNA) stained chromosomes digested with AluI or PvuII in patterns resembling those produced with Giemsa stain. These results support the theory that restriction endonucleases produce bands by cutting the DNA at specific base pairs and the subsequent removal of the fragments results in diminished staining by Giemsa. This technique is simple, reproducible, and in rainbow trout produces a more distinct pattern than that obtained with conventional C-banding methods.     

Karyotype of the gibbons hylobates lar and h. moloch inversion in chromosome 7.

A karyotype of the gibbon, Hylobates, has been prepared based on the chromosome banding patterns produced by quinacrine, trypsin-Giemsa, and centromeric heterochromatin stains. The banding patterns of H. lar and H. moloch are virtually identical. No brilliant quinacrine-fluorescent areas are present. The banding pattern of most of the gibbon chromosomes show less resemblance to those of the human, chimpanzee, gorilla, or orangutan than the chromosomes of the higher primates do to each other, suggesting a relatively large evolutionary separation of the gibbon from the higher primates. A pericentric inversion of chromosome 7 is present in one gibbon.     

Identical giemsa banding patterns of two macaca species: Macaca mulatta and M. fascicularis. A densitometric study.

Karyotypes of Macaca mulatta and M. fascicularis (2n equals 42) were compared using the Giemsa banding pattern technique. No differences between the chromosomes of both Macaca species were found with conventional staining methods. In the literature the occurrence of fertile hybrids is used as an argument in favor of conspecificity. The banding patterns were evaluated by comparison of their density distribution curves. Our data indicated no significant differences between the two Macaca species.     

Improved definition of chromosomal breakpoints using high-resolution multicolour banding

Characterisation of chromosome rearrangements using conventional banding techniques often fails to determine the localisation of breakpoints precisely. In order to improve the definition of chromosomal breakpoints, the high-resolution multicolour banding (MCB) technique was applied to identify human chromosome 5 breakpoints from 40 clinical cases previously assessed by conventional banding techniques. In 30 cases (75%), at least one breakpoint was redefined, indicating that MCB markedly improves chromosomal breakpoint localisation. The MCB pattern is highly reproducible and, in contrast to conventional banding pattern, is consistent in both short and elongated chromosomes. This might be of fundamental interest for the detection of chromosomal abnormalities, especially in tumour cells. Moreover, MCB even allows the detection of abnormalities that remain cryptic in GTG-banding analysis.     

Replication banding in the chromosomes of the European eel (Anguilla anguilla)

The chromosomes of the European eel Anguilla anguilla have been analyzed with a replication banding technique from lymphocyte cultures treated with 5-BrdU. This technique allows us to identify with high resolution the individual chromosome pairs and to differentiate classes of chromatin by the order of replication. The replication banding obtained on the chromosomes of European eel can be related with the structural bands described in this species.