Structural differentiation of the meiotic and mitotic chromosomes of the salamander,Ambystoma macrodactylum

The lampbrush chromosomes of the long-toed salamander, Ambystoma macrodactylum Baird, have been analysed and a map of the oocyte genome prepared. The location of C-bands and cold-induced-constrictions has been established in mitotic chromosomes and compared with the location of marker structures and chiasmata in several lampbrush bivalents. In the lampbrush chromosomes, C-bands are tentatively correlated with sphere-organizing loci and with regions of low chiasma frequency; cold-induced-constrictions are tentatively correlated with regions of high chiasma frequency. In general, in this salamander, C-bands do not coincide in position with cold-induced-constrictions. We have compared our results with those obtained by Callan (1966) in his investigation of chromosomes of the axolotl, Ambystoma mexicanum, and we present an analysis of the similarities and differences that are visible in the chromosome sets of these two ambystomatid species.     

C-band proximity of chiasmata and absence of terminalisation in Allium flavum (Liliaceae)

Chiasmata in male meiosis of Allium flavum from early diplotene to metaphase I are found to occur almost exclusively very close to the terminal and intercalary C-bands. Heterozygosity of intercalary C-bands, where one homologue has one band missing, allows the direct proof that chiasmata do not terminalise and therefore mark the point of exchange.     

Studies on chiasma distribution and chiasma movement in Zoniopoda tarsata (Orthoptera: Romaleidae)

A sample of 27 males of Zoniopoda tarsata from Argentina was studied cytologically. The three largest autosomal pairs and the X were characterized by the presence of interstitial C-bands. Chiasma position relative to the bands was analyzed at diplotene and diakinesis. The frequency of interstitial, terminal and total chiasmata per cell was studied for the whole autosomal bivalent set, analysing the variations between stages and among individuals. The comparison of interstitial chiasma frequencies between stages and among individuals and the study of chiasma position relative to the bands in pairs 1, 2 and 3 indicated that chiasma distribution varied from diplotene to diakinesis. Therefore, terminalization does exist in this species and the movement may occur towards the centromere. The frequency of terminal associations at diplotene showed a high negative correlation (r=-0.89; p<10^-5) with the number of interstitial chiasmata. This correlation would not be expected if the two kinds of association were produced by different (independent) mechanisms. Consequently, terminal associations were considered genuine chiasmata. The correlation between interstitial and total chiasmata was very much lower then the former (r=0.39; p=0.04). This fact, besides the relatively low variation for chiasma number, observed among individuals suggests that in this species the number of interestitial chiasmata, which are the most important in controlling the genetical recombination, is mainly regulated by changes in chiasma distribution, while variations in total chiasma frequency are of much lower magnitude.Member of Carrera del Investigador del Consejo Nacional de Investigaciones Cientificas y Técnicas (CONICET)     

Orientation of interphase chromosomes as detected by Giemsa C-bands

The orientation of Giemsa C-bands has been studied in mitotic and interphase cells of Allium cepa, A. sativum and of Aloe vera. The C-bands in these three species are located at the telomeres, secondary constriction region of the nucleolar chromosomes and the centromeric regions, respectively. Observations in A. cepa and Aloe indicate clearly that the interphase chromosomes are non-random in their orientation and possibly maintain their telophase configuration through the attachment of telomeres and perhaps of kinetochores with the nuclear membrane. Electron micrographs of onion cells also reveal that certain heterochromatic segments are associated with the nuclear membrane. — The nucleolar interstitial C-bands in A. sativum remain free in the nucleoplasm and may come close to each other due to heterochromatic attraction. Such a heterochromatic attraction is also evident between telomeric regions and between centromeres. However, a two by two attachment could not be noticed. A diagrammatic representation of the orientation of interphase chromosomes has been presented.The major part of this work was presented at the First International Congress on Cell Biology, Boston, Sept. 5–10, 1976 (Platform Session 36, J. Cell Biol. 70, 418a (1976)     

Effects of supernumerary heterochromatin on chiasma condition in two species of Acrididae (Orthoptera)

Amblytropidia australis and Dichroplus elongatus were found to be polymorphic for supernumerary heterochromatin. In both, basic karyotypes are 2n=22+XO in males.Mitotically unstable extra chromosomes were detected in a population of A. australis. The Bs are telocentric and their number varies from O to 2 within individuals. Mean frequencies of interstitial and total chiasmata at diplotene were compared between individuals with and without Bs. The mean frequency of interstitial chiasmata increases with the number of Bs per cell.A supernumerary terminal segment in S₁₀ pair was observed in a heterozygous condition in several individuals of D. elongatus. The localization and frequency of chiasmata at diplotene were studied. The segment has an intrachromosomal effect since it modifies the location of chiasmata in the bivalent involved.Fellow of the Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET).     

Different replication patterns of chromocentres and C-bands inLilium henryi

To determine if interphase chromocentres are fully equivalent to mitotic C-bands in plants, their times of replication have been compared in the large genome (1C=35 pg) ofLilium henryi. Nuclei of the root-tip cortex were pulse labelled with³H-thymidine and labelling patterns carefully followed in semi-thin sections during a 12 h chase period. Chromocentres decondense and replicate in the later stages of S-phase, after euchromatin has completed its replication. Late-replicating regions, reflecting a portion of the chromocentric material, were then mapped in mitotic chromosomes and found to be localized to the sub-distal and distal regions of all long chromosome arms. Most of the chromatin in these regions is non C-banded and, further, not all C-bands are located here. Some of the 11 inter-calary and 2 nucleolar C-bands are found in earlier replicating regions, as are the 12 centric bands. ThereforeLilium C-bands do not all replicate at the end of S-phase. Chromocentres occupy 17–18% of interphase nuclear volume while C-bands make up only 3.7% of the area of mitotic chromosomes. We conclude thatLilium chromocentres contain much other chromatin in addition to C-bands, and therefore that chromocentres and C-bands cannot be universally equated.     

Cytological mapping of the M and D loci in the mosquito,Aedes aegypti (L.)

The separate identities of the male-determining factor, M, and the sex-linked Distorter gene, D, are established in an Accra strain of Aedes aegypti. Their to each other and to Giemsa C-bands. Thus, M is invariably inherited with the centromere, whereas D lies towards the intercalary band. Approximately 1.2% recombination occurs between M and D but, in a chromosome known for its distal localization of chiasmata, it is argued that he two are not necessarily as closely linked cytologically as this might imply. Evidence on the genetic effects of recombination in the region of M and D is also considered.The work was supported by a grant from the Science Research Council.     

Mitotic chiasmata and other quadriradials in mitomycin C-treated Bloom's syndrome lymphocytes

Mitotic chiasmata and other quadriradials (QRs) were studied by Q-banding in mitomycin C-treated and untreated lymphocytes from two sibs with Bloom's syndrome. The frequency of chiasmata was very significantly increased by the mitomycin treatment in cells from both sibs. Chiasmata occurred throughout the chromosomes, but were favored in Q-dark regions, particularly at borders between dark and light regions (Kuhn, 1976). No significant difference was found in the distribution of chiasmata among chromosome regions in treated and untreated material. This differs from the reported action of mitomycin C on cultured lymphocytes of normal persons, where chiasmata are concentrated at secondary constrictions and centromeres. Adjacent counterparts to mitotic chiasmata, and chromatid translocations between non-homologous chromosomes, also occurred in the treated material, but with a much lower frequency than mitotic chiasmata. This again differs from the effects of mitomycin C on lymphocytes of normal persons, where chiasmata account for 20% or less of total QRs.This is paper No. 2054 from the Genetics Laboratory, University of Wisconsin, Madison     

C-banding analysis on wild Emmer (Triticum dicoccoides Körn) strains with and without spontaneous reciprocal translocations

C-banding polymorphism was analyzed in eight strains of wild Emmer, Triticum dicoccoides Körn, which included six translocation homozygotes reported previously. Polymorphisms were detected in all of the strains examined, and the breakpoints of five spontaneous translocations were successfully identified by C-bands. Of the eight breakpoints that could be precisely identified, one was located in the centromeric region while the remaining seven were located in proximal to distal euchromatic regions. The two breakpoints of one translocation could only be approximately localized to proximal regions due to the scarcity of C-bands. The present results are in contrast with those observed on T. araraticum, another wild tetraploid wheat belonging to the Timopheevi group, in which most of the breakpoints were located in centromeric regions. In T. dicoccoides, the six translocation chromosome types were derived from the standard karyotype primarily by a mechanism other than centric breakage-fusion.     

Late labelled regions in relation to Q- and C-bands in chromosomes of Lilium longiflorum and L. pardalinum

Root tips were pulse-labelled with tritiated thymidine. Late-labelled regions were mapped by quantitative autoradiography of metaphase chromosomes collected 11 h after the pulse for longiflorum (mean G₂=14 h), and 13 h for pardalinum (mean G₂=18 h). Late label in both species was preferentially located in sub-distal regions of the longer chromosome arms. Minimal labelling occurred in centromeric areas. — Some brightly Q-banded regions were late labelled, and some dull areas were not. However, late patterns were considerably more localised than bright Q-bands, and late regions were closely similar between species whereas Q-band patterns are not. Therefore bright Q-bands are apparently not consistently late replicating in Lilium, as they are in mammals, and they may therefore represent a different category of chromosomal substructure. — Centromeric C-bands and those at most nucleolar organisers were not late labelled. Only the more distal intercalary C-bands replicated late, and they were not significantly later than the chromatin surrounding them.     

Mitotic and meiotic analysis of a pericentric inversion associated with a tandem duplication in Eleutherine bulbosa

The mitotic and meiotic chromosomes of five populations of Eleutherine bulbosa were analysed after C-banding and fluorochrome (DAPI/CMA) staining. All individuals showed heterozygosity in pair number I for a pericentric inversion and a tandem duplication in the inverted region of one of the homologues. The duplication comprised the secondary constriction and ca. 40% of its associated CMA⁺ heterochromatin. All the secondary constrictions were nucleolar organizers, but the repeated one seemed to be the most active. The inverted segment constituted ca. 70% of the chromosome length and included, in addition to the duplication and the centromere, two DAPI⁺ C-bands. These markers, especially the CMA⁺ blocks which were unique in the complement, permitted a detailed analysis of meiotic pairing. In practically all examples of late zygotene the CMA blocks were paired, which seems to mean that non-homologous pairing never occurred. At this stage the duplicated CMA⁺ block was clearly unpaired. At pachytene-diplotene the inverted region formed a typical loop without chiasmata. Although male meiosis was normal, no indication of sexual reproduction was found. A causal relationship between duplication, inversion and asexual reproduction is proposed to explain the maintenance of the heterozygosity.     

Karyotype of Mesembryanthemum crystallinum (Aizoaceae) studied by chromosome banding,FISH with rDNA probes and immunofluorescence detection of DNA methylation

The karyotype of the common ice plant Mesembryanthemum crystallinum L. (Aizoaceae) was studied using Chromomycin A₃ (CMA)/4′,6-diamidino-2-phenylindole (DAPI) staining, fluorescence in situ hybridization with 5S and 18S–5.8S–25S rDNA probes, DAPI/C-banding and immunodetection of 5-methylcytosine. A single bright CMA-band was revealed on the satellite chromosome, whose location was coincided with a position of a site of 18S–5.8S–25S rRNA genes. A site of 5S rRNA genes was observed on one of the other chromosomes. Relatively large DAPI/C-bands were mainly localized in the pericentromeric regions of the chromosomes. DAPI/C-banding patterns allowed us to identify all the chromosomes in the karyotype of M. crystallinum. The methylation of euchromatic chromosome regions was weaker as compared with heterochromatic DAPI/C-bands, which were hypermethylated. The obtained results may provide opportunities for investigating, at the chromosomal level, the genomic changes occurring in M. crystallinum either under salinization or under the action of other stress factors.     

Q-bands in Lilium and their relationship to C-banded heterochromatin

In L. pardalinum, narrow bands of quinacrine fluorescence are distributed throughout the chromosomes. These vary in intensity from dull to bright, and their constant pattern allows all chromosomes to be recognized. Bright bands occur at some centromeres, and near all three nucleolar constrictions. In L. longiflorum, similar Q-bands occur along chromosomes, but they are less distinctive and their pattern does not closely match that of L. pardalinum. Also, L. longiflorum does not have bright regions at or near primary and secondary constrictions. Most Q-bands do not coincide with dark Giemsa C-bands, except for the bright nucleolar and centromeric regions in L. pardalinum. All C-banded heterochromatin stains identically after SSC pretreatment, dark with Giemsa and bright with quinacrine.— The many Q-bands of varying intensity, wide distribution and constant pattern, unrelated to C-bands, may be analogous to mammalian Q-bands. Such universality is expected if Q-bands area fundamental component of chromosome architecture.     

The meiotic behaviour of autosomal heterochromatic segments in hedgehogs

Male meiosis in the two species of hedgehogs Erinaceus europaeus and Aethechinus algirus, possessing respectively three and two pairs of autosomes with large blocks of heterochromatin, has been studied. The heterochromatic segments pair homologously till the end of pachytene, but separate during diplotene, owing to lack of chiasmata in these regions. They also organize the nucleolus in both species. The sex chromosomes (sex vesicle) are not associated with the nucleolus. The lack of chiasmata in the heterochromatic segments is interpreted as possible mechanism for the conservation of vital genes, such as ribosomal cistrons.     

Chromosome polymorphism in the Italian newt,Triturus italicus

A chromosomal variation, changing shape and C-banding pattern of chromosome XII of Triturus italicus was detected among the offspring of two F₁ hybrid families of T. italicus × T. vulgaris meridionalis . In both families a number of individuals appeared to have a metacentric instead of the expected subtelocentric chromosome XII of T. italicus. — Investigations in three well separated localities in the range of the species showed the polymorphism to have a wide distribution and to be part of a complex pattern involving at least two inversions and (presumably) deficiencies of large C-bands. At meiosis, the shape of bivalent XII, and the location and frequency of chiasmata in the bivalent varied with the karyomorph involved. It is suggested that large rearrangements may still play an important role in the karyological evolution of Triturus.     

Comparative chiasma analysis using a computerised optical digitiser

A new computerised technique has been devised for measuring the distribution of chiasmata along diplotene bivalents. The method involves the introduction into the field of view of the microscope, of a fine light spot which can be accurately manipulated along the chromosomes of each bivalent. The data recorded include (a) the positions of the chiasmata along the bivalent in terms of their relative distances from the centromere and (b) the individual bivalent and cellular chiasma frequencies. — The method has been applied to the analysis of chiasma distribution patterns in the two known species of the genus Caledia, C. species nova 1 and C. captiva and in two chromosomal races of the latter. Statistical tests indicate that within bivalents at least 40% of the comparative distribution patterns of chiasmata between races and species are significantly different. Similar comparisons between populations within races reveal only 18% significant differences. — The observed distribution patterns of chiasmata in this genus suggest that chiasma formation is sequential from centromere to telomere. — The variation in the frequency and distribution of chiasmata between races and species suggests that the interference distances between successive chiasmata are, at least partially, independent of chiasma frequency and position. — The interracial and interspecific differences in chromosome structure are correlated with changes in chiasma pattern.     

Banding patterns of the chromosomes of Presbytis cristatus pyrrhus and P. obscurus

The G- and Q-bands and the location of the nucleolar organizer regions (NOR) in the chromosomes of Presbytis obscurus and the Q- and C-bands of P. cristatus pyrrhus are described. Their chromosomes are compared to those of Macaca mulatta and to other Cercopithecidae and Hylobatidae. The origin of the two different banding patterns of pair no. 1 in our specimen of P. cristatus pyrrhus is discussed.     

The organization of DNA in the mitotic and polytene chromosomes of Sciara coprophila

The organization of DNA in the mitotic metaphase and polytene chromosomes of the fungus gnat, Sciara coprophila, has been studied using base-specific DNA ligands, including anti-nucleoside antibodies. The DNA of metaphase and polytene chromosomes reacts with AT-specific probes (quinacrine, DAPI, Hoechst 33258 and anti-adenosine) and to a somewhat lesser extent with GC-specific probes (mithramycin, chromomycin A₃ and anticytidine). In virtually every band of the polytene chromosomes chromomycin A₃ fluorescence is almost totally quenched by counterstaining with the AT-specific ligand methyl green. This indicates that GC base pairs in most bands are closely interspersed with AT base pairs. The only exceptions are band IV-8A3 and the nucleolus organizer on the X. In contrast, quinacrine and DAPI fluorescence in every band is only slightly quenched by counterstaining with the GC-specific ligand actinomycin D. Thus, each band contains a moderate proportion of AT-rich DNA sequences with few interspersed GC base pairs. — The C-bands in mitotic and polytene chromosomes can be visualized by Giemsa staining after differential extraction of DNA and those in polytene chromosomes by the use of base-specific fluorochromes or antibodies without prior extraction of DNA. C-bands are located in the centromeric region of every chromosome, and the telomeric region of some. The C-bands in the polytene chromosomes contain AT-rich DNA sequences without closely interspered GC base pairs and lack relatively GC-rich sequences. However, one C-band in the centromeric region of chromosome IV contains relatively GC-rich sequences with closely interspersed AT base pairs. — C-bands make up less than 1% of polytene chromosomes compared to nearly 20% of mitotic metaphase chromosomes. The C-bands in polytene chromosomes are detectable with AT-specific or GC-specific probes while those in metaphase chromosomes are not. Thus, during polytenization there is selective replication of highly AT-rich and relatively GC-rich sequences and underreplication of the remainder of the DNA sequences in the constitutive heterochromatin.     

Prometaphase chromosomes of the howler monkey (Aloutta caraya): G,C, NOR,and restriction enzyme (RES) banding

Prometaphase lymphocyte chromosomes from eight adult argentinian Alouatta caraya females were characterized using sequential G-C banding techniques, Ag-NOR bands and bands obtained with the restriction enzymes Hae III, Eco RI, Alu I and Sau 3A. The cytogenetic analysis showed 2n = 52, with four, five, or six NOR chromosomes. Digestion with Hae III and Eco RI produced G-like-bands. Centromere regions and two interstitial C-bands (in chromosomes number 16 and 21) showed intraindividual or interindividual heterochromatic polymorphisms. Alu I digestion produced C-like bands with gaps in the centromere regions, and Sau 3A produced C-like bands. The karyotypes and banding patterns of A. caraya, A. palliata, A. belzebul, and A. seniculus are compared, based on whole chromosome and whole arm homeologies. © 1994 Wiley-Liss, Inc.     

Recombination nodules and chiasma localization in two orthoptera

The distribution of recombination nodules (RNs) is reported from observations on two-dimensional spreads of Locusta migratoria and Chloealtis conspersa spermatocytes; C. conspersa is a known example of a species with terminally localized chiasmata, while L. migratoria has nonspecific positioning of chiasmata. Measurements of the distances from 102 RNs to the ends of the synaptonemal complexes (SCs) on which they were found show the RNs to be near-terminally localized in C. conspersa and to occur along the lengths of the SCs in L. migratoria. Thus, the localization of RNs appears to reflect the localization of chiasmata. These observations are interpreted as support for the proposed recombinant function of RNs.