Meiosis in a sterile male mouse with an isoYq marker chromosome

A male mouse with a metacentric Y chromosome of twice the normal size has been studied chromosomally in bone marrow mitoses, spermatogonial mitoses, and diakinesis-metaphase I primary spermatocytes. A low frequency of nondisjunction for this chromosome (2%) was noted in both bone marrow and spermatogonial mitoses. In spermatogonial mitoses, loss of the Y chromosome had occurred to the extent that 12% of spermatogonia were XO, resulting in 17% XO primary spermatocytes. Hardly any stages beyond the primary spermatocyte stage were encountered, which agrees with testis weights of approximately 30% of normal. Surface-spread pachytene spermatocytes yielded few cells that were analyzable for their total complement of synaptonemal complexes. The Y chromosome showed complete fold-back pairing and was located far away from the X chromosome. X and Y chromosomes were paired in 14.5% of the diakinesis-MI spermatocytes that contained a Y chromosome. The origin of this chromosome is discussed against the background of localization of the gene for the testis-determining factor on the short arm of the mouse Y chromosome.     

A radiation analysis of a comstockiella chromosome system: destruction of heterochromatic chromosomes during spermatogenesis in Parlatoria oleae (Coccoidea: Diaspididae)

In the males of the olive scale insect, Parlatoria oleae (2n=8), the paternal set of chromosomes becomes heterochromatic during late cleavage or early blastula and remains so until spermatogenesis. Immediately before the onset of meiosis in the males one or more heterochromatic chromosomes disappear from each primary spermatocyte. At prophase four euchromatic and from one to three heterochromatic chromosomes are present in each cell. The disappearance of the heterochromatic chromosomes before meiosis could be due either to the dehetero-chromatization of the heterochromatic chromosomes and their subsequent pairing with their euchromatic homologues, or to the destruction of the heterochromatic chromosomes. — The alternative interpretations of spermatogenesis in P. oleae were tested by using chromosome aberrations, which had been induced in the heterochromatic set by paternal X-irradiation, as genetic markers in breeding tests of about 400 X₁ males. Meiosis was examined in X₁ males which showed conspicuous chromosomal rearrangements in their somatic cells. The absence of either heteromorphic chromosome pairs or multivalents at spermatogenesis and the failure of the X₁ males to transmit any form of chromosome aberration induced by paternal irradiation is strong evidence that the heterochromatic chromosomes are destroyed in P. oleae. — The evolutionary relationships of the chromosome systems in the coccids are considered. Models are outlined for the derivation of a Comstockiella system involving chromosome destruction either from a lecanoid sequence or from a hypothetical Comstockiella sequence involving chromosome pairing. Problems concerning the control of chromosome destruction are discussed.From a dissertation submitted in partial fulfillment of the requirements of Doctor of Philosophy in Genetics.This work was supported by grant GB 8196 from the National Science Foundation to Dr. Spencer W. Brown, and by a National Institutes of Health Fellowship 1 F02 CA 44173-01 to the author from the National Cancer Institute.Dedicated to Dr. Sally Hughes-Schrader on the occasion of her seventy-fifth birthday.     

Chromosome distribution and catenation in Rhoeo spathacea var. concolor

The twelve chromosomes of Rhoeo spathacea variety concolor are arranged in a definite sequence in a ring at meiosis. Identification of all the 12 chromosomes was possible in 119 diakinesis and metaphase I cells. — Pollen viability was measured to be 36.54% by cotton blue staining procedure. Forty five of 56 metaphase I cells (80.36%) had adjacent distribution. Each of the 12 chromosomes was equally likely to be involved in adjacent distribution regardless of their sizes and heterobrachialness. Adjacent distribution occurred randomly at each arm-position in the ring regardless of the lengths of the arm-pairs. — The most frequent chromosome configuration at diakinesis and metaphase I was a chain-of-12 chromosomes (41.18%). Cells with 1 to 4 chains of chromosomes were observed. The observed frequencies of various configurations were in good agreement with the calculated frequencies. The mean number of chiasmata was 10.90 per cell and 0.908 per pair of chromosome arms. The 131 chiasma failures were distributed at random among the 12 arm-positions. Since the lengths of arm-pairs in the ring vary, the randomness may mean that chiasma formation was limited to short terminal segments on all chromosomes.     

Production and identification of primary trisomics in diploid Agropyron cristatum (crested wheatgrass).

Six of the seven possible primary trisomics in Agropyron cristatum were produced. Based on morphology, arm length ratios, and C-banding patterns, they were identified as primary trisomics for chromosomes A, B, C, D, E, and G. Agropyron cristatum is one of several species constituting the crested wheatgrass complex. All species in this complex contain one basic genome (P). A study was conducted to produce and identify a primary trisomic series that will be used to map genes to individual chromosomes. A population of 157 plants were generated by crossing autotriploids (PPP) with diploid (PP) A. cristatum: 58 were diploid (2n = 14), 76 were primary trisomies (2n = 15), 17 were double trisomic (2n = 16), 4 were triple trisomics (2n = 14 + 3), 1 was telocentric trisomic (2n = 14 + 1 telo), and 1 was tetratrisomic (2n = 14 + 4). Karyotype analysis of acetoorcein-stained chromosomes was carried out using the CHROMPAC III computer program; for analysis of C-banded karyotypes, the computer imaging analysis program PCAS (Plant Chromosome Analysis System) was used to identify the primary trisomics. Of the 47 primary trisomics analyzed, 21 plants had one extra satellited chromosome E, 18 with the satellited D chromosome, 3 each for chromosomes B and G, and 1 each for chromosomes C and A. Chromosome pairing was studied in trisomies B, D, E, and G. Trisomics for chromosomes B and G were similar in their mieotic behavior. Each had a trivalent frequency of about 60% and pollen stainability of less than 40%. Trisomics for chromosomes D and E had a trivalent frequency of about 30% and pollen stainability of over 70%.     

Cytogenetics of inherited partial sterility in three generations of the large milkweed bug as related to holokinetic chromosomes

AdultOncopeltus fasciatus males were irradiated with 9000 R of X-rays, and crossed to untreated females. Fertility was reduced to 4.1%. F₁ males and F₂ and F₃ males and females were outcrossed to untreated partners. All F₁ males were partially or totally sterile and a significant number of F₂ and F₃ males and females had reduced fertility. The fertility of each generation was higher than the preceding one, even though the progeny studied in the 3rd generation were selected mostly from low-fertility lines. Cytogenetic studies showed that complex chromosome rearrangements and fragments were transmitted to each generation and were severe enough to account for reduced fertility. — The transmission of complex chromosome rearrangements and fragments for 3 generations of outcrossing correlates with the persistence of sterility in this species possessing holokinetic chromosomes. — Over half the inviable embryos derived from irradiated sperm from P₁ males died in the early stages of development. The inviable embryos produced in later generations died in much later stages of development. — A stable rearrangement of a Y-chromosome fragment translocated to an autosome was isolated from a single F₁ male. This rearrangement was transmitted to all F₂ and F₃ sons. Fertility of the males of this line was reduced to about 75–80%.Supported in part by AEC Contract No. AT(49-7)3028.     

Studies of multipolar mitoses in euploid tissue cultures

Cultures of kidney epithelium and fibroblasts of 39 specimens of Microtus agrestis were investigated. In all 77 cultures multipolar mitoses were found. They were studied in living state and after pulse labelling with ³H-thymidine. The ploidy of the multipolar mitoses and of their daughter nuclei was determined by measuring the relative Feulgen-DNA content and by counting the predominantly constitutive heterochromatic sex chromosomes. Constitutive heterochromatin was demonstrated by late replication, retarded separation of the chromatids in anaphase, heteropycnosis and by the Giemsa technique of Arrighi and Hsu (1971). The latter stained also the spindle apparatus of mitoses.—In living cells, transformation of multipolar mitoses into bipolar mitoses was observed. The chromosomes of multipolar mitoses are separated into complete genomes; the daughter nuclei can be haploid, diploid, triploid or tetraploid. The chromosomes of haploid and triploid metaphases were studied with the Giemsa banding technique. The banding pattern shows an exact monosomy and trisomy, respectively, for each chromosome. Haploid nuclei are likely to be viable only in multinucleate cells, whereas triploid cells behave like diploid cells during the S period and the mitosis.Dedicated to Prof. Dr. K. Goerttler on the occasion of his 75th birthday.Supported by the Bundesministerium für Bildung und Wissenschaft of the Federal Republic of Germany.     

The effect of cyclophosphamide on the centromere separation sequence in Chinese hamster spermatogonia

Summary The sequence of centromere separation in spermatogonial chromosomes of untreated and cyclophosphamide-treated Chinese hamsters is described.Centromeres of chromosome 1 and 2 separated much earlier than all other chromosomes, especially 6–8. Cyclophosphamide significantly inhibits the centromere separation in all chromosome groups but does not alter the sequence of separation.     

Occurrence of premature chromosome condensation in mouse spermatogonia treated with busulfan

Busulfan induction of premature chromosome condensation (PCC) was examined in mouse spermatogonia. Adult (C3H X SWV) F1 male mice were injected intraperitoneally with busulfan at 10 or 30 mg/kg of body weight and killed 18-72 h later. Polyploid-like mitoses were frequent (ca. 1/4 of all spermatogonial mitoses examined) in both the untreated and treated groups. Most of these were considered to have been derived from normal spermatogonia. In busulfan-treated mice, polyploid-like mitoses with PCC were seen. The frequency of PCC-containing mitoses increased with increasing dose and exposure time. A possible interpretation for PCC induction in mouse spermatogonia is discussed.     

Chromosomal instability in Lathyrus sativus L.

Summary In Lathyrus sativus (2n=14), variety LSD-1 shows an instability of somatic chromosome number which can be observed in root tip and shoot tip mitoses. In this variety, approximately 54% of the seedlings showed intra-individual variation in chromosome number ranging from 2n=14–3. This variability in chromosome number was recorded in approximately 60% of the dividing cells. Two seedlings were triploid with 21 chromosomes. Variation in chromosome number in somatic cells within individual plants is possibly controlled by genetic factors, which result in spindle abnormalities, chromosome degradation and minute chromosomes. The variation in chromosome number is probably responsible for the pollen polymorphism noted in this particular strain. The possible mechanism of intra-individual variability and the occurrence of the phenomenon vis-a-vis its applications are discussed.     

The chromosomes of tufted deer (Elaphodus cephalophus)

Mitotic and meiotic chromosome preparations of the tufted deer (Elaphodus cephalophus) were studied to elucidate the sex-chromosomal polymorphism evidenced by this species. Females had 2n = 46 or 47 chromosomes, whereas males had 2n = 47 or 48 chromosomes. An X;autosome translocation was identified by synaptonemal complex analysis of spermatocytes at pachytene and confirmed by the presence of a trivalent at diakinesis/metaphase I. The present work, in combination with earlier observations by others, indicates that E. cephalophus possesses a varied X-chromosome morphology involving an X;autosome translocation and addition of varying amounts of heterochromatin. It is speculated that sex-chromosome polymorphism may be responsible for the observed differences in diploid chromosome number of tufted deer.     

Vergleich der Chromosomensätze von Steinwild (Capra Ibex) und Hausziege (Capra Hircus)

Capra ibex has a diploid set of 60 chromosomes. They are all acrocentric. The Y chromosome is the only element that can be recognized individually in all mitoses. It is distinctly smaller than any autosome and its chromatids are not spread apart as is characteristic of all the other chromosomes. In these features Capra ibex and C. hircus are identical. The idiograms of the two species show that the chromosomes 18 to 21 and 41 to 44 differ, but not significantly, from each other. The Karyotypes of C. ibex and C. hircus are so alike, that the observed fertility of the hybrids does not surprise. — The amount of chromosome spiralisation does not influence the relative chromosome length. — The second longest chromosome in the haploid set of C. ibex is the X chromosome. It was taken into account that the length of the male set is shorter compared with the female set by the difference X minus Y.     

Nuclear conditions in haploid Pelargonium in vivo and in vitro

In the shoot apices of the haploid Pelargonium cultivar Kleine Liebling, all mitoses are haploid (n = 9); however, ca. 20% of the interphase nuclei have DNA contents greater than 2C (up to 4C), indicating a tendency to chromosome endoreduplication in this material. — In internodes in vivo, the few mitoses present are haploid (quite probably, cambium cells); in addition to haploid interphases (1C to 2C DNA contents), endoreduplicated (endopolyploid) nuclei and nuclei in the course of endoreduplication occur with a frequency of ca. 40–50% (DNA contents up to 8C). — When internodes are cultured in vitro, differentiated cells are stimulated to divide, thus forming a population of diploid and tetraploid mitoses in addition to the preexistent meristem (haploid) cell population. In the process of time, diploid and tetraploid mitoses continue to be present in the callus, whilst haploid mitoses may decrease in number and eventually disappear. All mitoses analyzed had euploid chromosome numbers (9, 18 and 36) and their DNA contents were correspondingly 2C, 4C and 8C. Since no extensive chromosome counts were made, aneuploidy in the cultured material cannot be excluded; but, if occurring, it should be rather rare. — Under the experimental conditions used, prolonged culture in vitro leads to the production of nuclei with DNA contents (16C and 32C) greater than those occurring in vivo (8C), due to one and two additional DNA replications respectively beyond the limits attained in vivo. Even in these cultures, however, a population of the meristematic haploid cell line (DNA values 1C to 2C) is still present. — The present results are discussed in their relations with previous works on nuclear conditions in vivo and in vitro and on regeneration processes in cultured tissues in plants.     

A complex sex-chromosome system in the hare-wallaby Lagorchestes conspicillatus Gould

Among specimens of the spectacled hare-wallaby Lagorchestes conspicillatus Gould (Marsupialia, family Macropodidae) 4 males had 15 chromosomes and 2 females 16 chromosomes. The sex chromosomes are X₁X₁X₂X₂ in the female and X₁X₂Y in the male, the Y being metacentric and both X chromosomes are acrocentric. In about 96% of sperm mother cells at meiosis the sex chromosomes form a chain trivalent and in more than 99% of these this orients convergently so that the X₁ and X₂ move to the same pole. Evidence is presented that L. conspicillatus has evolved from a form with 22 chromosomes including a small X and a minute Y. Autoradiographic studies show that the proximal fifth of the X₁ chromosome replicates late. This is probably the ancestral X chromosome which has been translocated to an autosome. The fate of the original Y is obscure but an hypothesis is proposed that it forms the centromeric region of the Y. A single male had 14 chromosomes and was heterozygous for a translocation involving the centric fusion of two acrocentric autosomes. In about 30% of sperm mother cells the autosomal trivalent did not disjoin regularly but, despite this, all secondary spermatocytes observed at metaphase 2 had balanced complements of chromosomes. It is assumed that unbalanced secondary spermatocytes died before reaching metaphase.     

Arrangement of prematurely condensed chromosomes in cultured cells and lymphocytes of the Indian muntjac

Premature chromosome condensation (PCC) was induced in order to study the arrangement of muntjac chromosomes in the interphase nuclei of proliferating and resting cells with respect to their polarity and the spatial relationship between them. The data were compared with the situation in in situ fixed and colcemid blocked metaphases. It appears that in rapidly dividing cells almost all G₁- and G₂ interphase chromosomes exhibit the Rabl type polarized orientation. This pattern still predominates in G₀ lymphocytes which may have been arrested at this stage for some months or even years. — The location of the small chromosome Y₂ was found to be central in normal metaphases but peripheral in colcemid blocked mitoses. The behavior in the premature condensed chromosome preparations was intermediate. Measurements of centromere distances between all possible pairs of chromosomes as well as on the relative position of chromosomes in circular spreads revealed no evidence for homologous somatic association during interphase and metaphase or any other suprachromosomal ordering principle. Interphase chromosome orientation seems to be solely the result of chromosome arrangement of the foregoing anaphase. Association between heterochromatic regions or the nucleolus organizers did not substantially influence this pattern. There is no support for speculations that in mammalian cells close proximity of homologoues sites is instrumental in functional cooperation.     

Chromosome distribution in a23 chinese hamster fibroblasts

This study deals with a systematic chromosome position analysis of 116 anti-mitotic and hypotonic treated a₂₃ chinese hamster cells. No chromosome or pair of chromosomes was found to be located nearer the center or the periphery of the metaphase plate than would be expected by the reference distribution. The homologous chromosomes of pair 2 lie nearer to each other but they do not form a specific angle. The same relative position was shown for the chromosome groups 1–2, 1-E1 and 2-E5 (E standing for an extra chromosome). On the other hand the chromosomes of the combinations X-7, X-8, 7–8, 8–11 and X-E2 were lying further from each other, while chromosomes 10-E1 had a greater mean angle. The non random distribution of the chromosomes 1 and 2 may be interpreted as function of their possibly more frequent participation in the organization of nucleoli. — To obtain more information about the influence of preparation techniques on the alteration of the chromosome position in metaphase plates, this study deals with some overall considerations about chromosome position. It is shown that in a₂₃ cells the smaller chromosomes do not tend to lie nearer the metaphase plate center (as it happens in human cells). Also a significant correlation between the chromosome position with respect to the metaphase plate center and the mean interchromosomal distances was not found in this type of cells.     

Autoradiographic studies of the human Y chromosome

An autoradiographic analysis (using continuous labeling with tritiated thymidine) was made on 317 cells from four normal males. The labeling pattern of the Y chromosome was compared to the first and the last chromosomes to complete replication as well as to G21–22. The Y chromosome was never found to be the last chromosome in the cell to complete replication. Instead, it completed DNA synthesis relatively early (usually among the first 10 chromosomes) but had a distinctively heavy label during the earliest stages of late-S. In 51% of those cells with one labeled G+Y chromosome, a G21–22 was labeled and the Y was not.—It was concluded, therefore, that the human Y chromosome is not a late-replicating chromosome but terminates replication earlier than most of the autosomes. In addition, the Y chromosome cannot be distinguished from the G chromosomes on the basis of a consistent and differential labeling pattern.Supported by USPHS Grant GM 15361.     

Cytological and molecular characterization of oat x maize partial hybrids

In cereals, interspecific and intergeneric hybridizations (wide crosses) which yield karyotypically stable hybrid plants have been used as starting points to widen the genetic base of a crop and to construct stocks for genetic analysis. Also, uniparental genome elimination in karyotypically unstable hybrids has been utilized for cereal haploid production. We have crossed hexaploid oat (2n=6x=42, Avena sativa L.) and maize (2n=2x=20, Zea mays L.) and recovered 90 progenies through embryo rescue. Fifty-two plants (58%) produced from oatxmaize hybridization were oat haploids (2n=3x=21) following maize chromosome elimination. Twenty-eight plants (31%) were found to be stable partial hybrids with 1–4 maize chromosomes in addition to a haploid set of 21 oat chromosomes (2n=21+1 to 2n=21+4). Ten of the ninety plants produced were found to be apparent chromosomal chimeras, where some tissues in a given plant contained maize chromosomes while other tissues did not, or else different tissues contained a different number of maize chromosomes. DNA restriction fragment length polymorphisms (RFLPs) were used to identify the maize chromosome(s) present in the various oat-maize progenies. Maize chromosomes 2, 3, 4, 5, 6, 7, 8, and 9 were detected in partial hybrids and chromosomal chimeras. Maize chromosomes 1 and 10 were not detected in the plants analyzed to-date. Furthermore, partial self-fertility, which is common in oat haploids, was also observed in some oat-maize hybrids. Upon selfing, partial hybrids with one or two maize chromosomes showed nearly complete transmission of the maize chromosome to give self-fertile maize-chromosome-addition oat plants. Fertile lines were recovered that contained an added maize chromosome or chromosome pair representing six of the ten maize chromosomes. Four independently derived disomic maize chromosome addition lines contained chromosome 4, one line carried chromosome 7, two lines had chromosome 9, one had chromosome 2, and one had chromosome 3. One maize chromosome-8 monosomic addition line was also identified. We also identified a double disomic addition line containing both maize chromosomes 4 and 7. This constitutes the first report of the production of karyotypically stable partial hybrids involving highly unrelated species from two subfamilies of the Gramineae (Pooideae — oat, and Panicoideae — maize) and the subsequent recovery of fertile oat-maize chromosome addition lines. These represent novel material for gene/ marker mapping, maize chromosome manipulation, the study of maize gene expression in oat, and the transfer of maize DNA, genes, or active transposons to oat.Joint contribution of the Minnesota Agricultural Experiment Station and USDA-ARS. Scientific journal series paper No. 21 859 of the Minnesota Agricultural Experiment Station. Mention of a trademark or proprietary product does not constitute a guarantee or warranty by the USDA-ARS or the University of Minnesota and does not imply approval over other products that also may be suitable     

TRISOMICS IN SOLANUM CHACOENSE: FERTILITY AND CYTOLOGY

Twenty trisomic plants found in the progeny 3x x 2x crosses in Solatium chacoense and their F₁ trisomies obtained by 2x + 1 X 2x crosses were studied with respect to their fertility and cytology. The female transmission of the extra chromosome in the trisomics varied from 2 to 60 %. The transmission frequencies of F₁ trisomies were similar to their parent trisomies in most of the lines. The transmission through the pollen ranged from 0 to 20 %. Female and male fertility of the parent trisomies was high. They produced an average of 37 seeds per pollination as the female or as the male parent. The F₁ trisomies produced about half the seed set of their parent trisomies. The extra chromosomes of six trisomies were identified by pachytene analysis. They were isochromosomes for the long arms of chromosomes I, IV and IX and the short arms of IV, IX and XII. Chromosome morphology of the extra chromosomes in pachytene stage was described. A chromosome association of 12 II + 1 I was found in 66 % of the cells at MI. About 29 % of the cells had one trivalent and 5 % had three or five univalents. The frequency of trivalent formation was not affected by the length of the extra chromosome. The possibility of univalent shift in secondary trisomies was discussed.     

Etude de la distribution générale des 46 chromosomes dans les cellules humaines en métaphase

Summary This study, based on the square distances of chromosome centromeres from the center of a cell, shows after circular transformation, that the general dispersion of the 46 chromosomes is uniform, except on the periphery of the cell. It is found that the chromosomes taken separately have not a random distribution, but have peculiar positions. In males, chromosomes 1, 13-14-15 and 21–22 tend to lie nearer the middle of the nucleus, while chromosomes, Y, 6-X and 4–5 tend to lie nearer the periphery. In female mitoses the chromosome 13-14-15, 21–22 and 19–20 and 19–20 have a more central position, while chromosomes 4–5, 3, 2 and 16 tend to lie nearer the periphery.

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Karyotype study on pseudoscorpions of the genus Lasiochernes Beier (Pseudoscorpiones, Chernetidae)

Karyotypes of the genus Lasiochernes (Pseudoscorpiones, Chernetidae) are studied for the first time. The diploid chromosome numbers of males were found to be 2n = 61 in L. pilosus, 2n = 69 in L. siculus and 2n = 73 in L. cretonatus. Karyotypes of all species mainly consist of biarmed chromosomes; the sex chromosome system is XO. Remarkably, the X chromosome displays partial (L. cretonatus), or even total (L. pilosus), negative heteropycnosis during the spermatogonial metaphase.