Chromosome structure and pairing preferences in autotetraploid rye (Secale cereale)

Genome ◽  
1994 ◽  
Vol 37 (5) ◽  
pp. 784-793 ◽  
Author(s):  
G. Jenkins ◽  
R. Chatterjee

The influence of chromosome structure upon pairing behaviour during meiosis was investigated by comparing four autotetraploid genotypes of rye (Secale cereale) containing homologous chromosome sets with different degrees of structural similarity. The series provided a range of genotypes that, at one extreme, contained structurally identical chromosome sets and, at the other extreme, sets that are certainly more heterozygous in the genic sense and probably also more diverse from a purely structural viewpoint. Relative frequencies of pairing configurations at meiotic prophase and metaphase I were compared by electron microscopy of whole-mount surface-spread synaptonemal complex complements and light microscopy of squash preparations. Despite unexpectedly low quadrivalent frequencies over all four genotypes, higher mean bivalent frequencies appeared to be associated with greater homologue diversity. In other words, greater structural divergence between chromosome sets appears to facilitate more efficient discrimination between homologous and identical chromosomes that drives the formation of bivalents. Statistical comparisons were not able to confirm in some cases the significance of the observed pattern of pairing behaviour.Key words: autotetraploid, rye, synapsis, chiasmata.

Author(s):  
Tadashi A. Okada ◽  
David E. Comings

Although many observations have been made on the ultrastructure of chromosomes by whole mount electron microscopy, the large size of most chromosomes has made it difficult to trace the chromatin fibers throughout the entire course of a given chromatid.In the present study, microchromosomes of the chicken were used to provide information on this facet of chromosome structure. Cells from a tissue culture of chicken embryo fibroblasts were exposed to colcemid 0.1 ug/ml for 3 hours and the cells in mitosis were selectively removed by shaking with 0.05% trypsin.


Genome ◽  
1992 ◽  
Vol 35 (6) ◽  
pp. 1054-1061 ◽  
Author(s):  
J. S. Rufas ◽  
J. L. Santos ◽  
M. Diez ◽  
J. A. Suja

The development of silver-stained synaptonemal complexes (SCs) and of chromatid cores was analyzed in squashed and surface-spread grasshopper spermatocytes using light and electron microscopy, respectively. This study was conducted to determine the relationship of the two chromosome structures and then obtain more insight into the meiotic chromosome structure. Pachytene cells observed by light microscopy showed thin silver-stained threads, representing SCs, along the centre of the bivalents. However, fully formed SCs, and an axial element corresponding to the univalent sex chromosome, appeared when these cells were observed by electron microscopy. During early diplotene no silver-stained threads were observed by light microscopy. However, fragmentation of the SCs was apparent in cells at the same stage when observed by electron microscopy. Both light and electron microscopy showed that chromosome cores were first detected in homologues of late diplotene – early diakinesis cells. During diakinesis the cores were not continuous but were interrupted where interstitial chiasmata occur. In prometaphase I – metaphase I cells these cores appeared continuous and double, i.e., each chromatid clearly showed its own core. We propose a model whereby the associated cores of sister chromatids act as frameworks for the formation of the SC lateral elements.Key words: meiosis, chromosome structure, synaptonemal complex, chromatid core.


Genome ◽  
1994 ◽  
Vol 37 (6) ◽  
pp. 1035-1040 ◽  
Author(s):  
A. L. del Cerro ◽  
A. Fernández ◽  
J. L. Santos

Meiotic pairing behaviour of one and two B isochromosomes (iso-Bs) in the grasshopper Omocestus burri was analysed by electron microscopy in surface-spread prophase I nuclei and compared with light microscopic observations of metaphase I. Iso-Bs display a peripheral location in the surface-spread nuclei and early pairing relative to that of the long members of the A set. Single iso-Bs undergo foldback pairing to give symmetrical hairpin loops. Two iso-Bs may show interarm pairing, mterchromosome pairing, or combinations of the two. Pericentromeric interarm pairing can be delayed in one or both Bs and this delay is mostly observed in bivalents with pairing partner switches. The iso-B bivalent frequencies observed in the three males analysed were 64, 44, and 41%, respectively; the two latter values were significantly lower than the 66% predicted by the random-end-pairing model. There is a reduction in the frequencies of iso-ring univalents (in 1B males) and bivalents (in 2B males) from pachytene to metaphase I. Similarities and differences between the pairing behaviour of iso-Bs from different species are also discussed.Key words: B isochromosomes, meiosis, grasshopper, synaptonemal complexes, pairing partner switches.


Author(s):  
David E. Comings ◽  
Tadashi A. Okada

Various aspects of chromosome structure based on whole mount electron microscopy and biochemical investigations will be discussed. The observations and conclusions include the following. 1. The appearance of half-chromatids is probably the result of an illusion of doubleness produced by the peculiar geometry of coiled structures. We suggest that their existence as an observation is perfectly valid, but the conclusion that they prove there are two independent portions of the chromatid (two DNA helices) is not. 2. The appearance of chromatin in whole mount preparations suggests that purely morphological observations will never convincingly prove whether chromosomes are single or multistranded. 3. Metacentric chromosomes possess a quadripartite centromere region composed of two areas of chromatin fiber association. These have presumably arisen from the fusion of two telocentric chromosomes each with only one area of chromatin fiber association. 4. Whole mount preparations of mitotic chromosomes frequently show the presence of nuclear membrane fragments attached to chromatin fibers at many sites along the full extent of the chromosome.


Author(s):  
H. Koike ◽  
S. Sakurai ◽  
K. Ueno ◽  
M. Watanabe

In recent years, there has been increasing demand for higher voltage SEMs, in the field of surface observation, especially that of magnetic domains, dislocations, and electron channeling patterns by backscattered electron microscopy. On the other hand, the resolution of the CTEM has now reached 1 ∼ 2Å, and several reports have recently been made on the observation of atom images, indicating that the ultimate goal of morphological observation has beem nearly achieved.


Author(s):  
J. H. Hayden

In a previous study, Allen video-enhanced constrast/differential interference constrast (AVEC-DIC) microscopy was used in conjunction with immunofluorescence microscopy to demonstrate that organelles and vesicle move in either direction along linear elements composed of microtubules. However, this study was limited in that the number of microtubules making up a linear element could not be determined. To overcome this limitation, we have used AVEC-DIC microscopy in conjunction with whole mount electron microscopy.Keratocytes from Rana pipiens were grown on glass coverslips as described elsewhere. Gold London Finder grids were Formvar- and carbon coated, and sterilized by exposure to ultraviolet light. It is important to select a Formvar film that gives a grey reflection when it is floated on water. A silver film is too thick and will detract from the image in the light microscope.


Author(s):  
D. R. Abrahamson ◽  
P. L. St.John ◽  
E. W. Perry

Antibodies coupled to tracers for electron microscopy have been instrumental in the ultrastructural localization of antigens within cells and tissues. Among the most popular tracers are horseradish peroxidase (HRP), an enzyme that yields an osmiophilic reaction product, and colloidal gold, an electron dense suspension of particles. Some advantages of IgG-HRP conjugates are that they are readily synthesized, relatively small, and the immunolabeling obtained in a given experiment can be evaluated in the light microscope. In contrast, colloidal gold conjugates are available in different size ranges and multiple labeling as well as quantitative studies can therefore be undertaken through particle counting. On the other hand, gold conjugates are generally larger than those of HRP but usually can not be visualized with light microscopy. Concern has been raised, however, that HRP reaction product, which is exquisitely sensitive when generated properly, may in some cases distribute to sites distant from the original binding of the conjugate and therefore result in spurious antigen localization.


Author(s):  
K.H. Westmacott

Life beyond 1MeV – like life after 40 – is not too different unless one takes advantage of past experience and is receptive to new opportunities. At first glance, the returns on performing electron microscopy at voltages greater than 1MeV diminish rather rapidly as the curves which describe the well-known advantages of HVEM often tend towards saturation. However, in a country with a significant HVEM capability, a good case can be made for investing in instruments with a range of maximum accelerating voltages. In this regard, the 1.5MeV KRATOS HVEM being installed in Berkeley will complement the other 650KeV, 1MeV, and 1.2MeV instruments currently operating in the U.S. One other consideration suggests that 1.5MeV is an optimum voltage machine – Its additional advantages may be purchased for not much more than a 1MeV instrument. On the other hand, the 3MeV HVEM's which seem to be operated at 2MeV maximum, are much more expensive.


Genetics ◽  
2003 ◽  
Vol 163 (4) ◽  
pp. 1337-1356 ◽  
Author(s):  
Adelaide T C Carpenter

Abstract The meiotic phenotypes of two mutant alleles of the mei-W68 gene, 1 and L1, were studied by genetics and by serial-section electron microscopy. Despite no or reduced exchange, both mutant alleles have normal synaptonemal complex. However, neither has any early recombination nodules; instead, both exhibit high numbers of very long (up to 2 μm) structures here named “noodles.” These are hypothesized to be formed by the unchecked extension of identical but much shorter structures ephemerally seen in wild type, which may be precursors of early recombination nodules. Although the mei-W68L1 allele is identical to the mei-W681 allele in both the absence of early recombination nodules and a high frequency of noodles (i.e., it is amorphic for the noodle phene), it is hypomorphic in its effects on exchange and late recombination nodules. The differential effects of this allele on early and late recombination nodules are consistent with the hypothesis that Drosophila females have two separate recombination pathways—one for simple gene conversion, the other for exchange.


Genetics ◽  
2001 ◽  
Vol 157 (3) ◽  
pp. 1387-1395 ◽  
Author(s):  
Sudhir Kumar ◽  
Sudhindra R Gadagkar ◽  
Alan Filipski ◽  
Xun Gu

AbstractGenomic divergence between species can be quantified in terms of the number of chromosomal rearrangements that have occurred in the respective genomes following their divergence from a common ancestor. These rearrangements disrupt the structural similarity between genomes, with each rearrangement producing additional, albeit shorter, conserved segments. Here we propose a simple statistical approach on the basis of the distribution of the number of markers in contiguous sets of autosomal markers (CSAMs) to estimate the number of conserved segments. CSAM identification requires information on the relative locations of orthologous markers in one genome and only the chromosome number on which each marker resides in the other genome. We propose a simple mathematical model that can account for the effect of the nonuniformity of the breakpoints and markers on the observed distribution of the number of markers in different conserved segments. Computer simulations show that the number of CSAMs increases linearly with the number of chromosomal rearrangements under a variety of conditions. Using the CSAM approach, the estimate of the number of conserved segments between human and mouse genomes is 529 ± 84, with a mean conserved segment length of 2.8 cM. This length is <40% of that currently accepted for human and mouse genomes. This means that the mouse and human genomes have diverged at a rate of ∼1.15 rearrangements per million years. By contrast, mouse and rat are diverging at a rate of only ∼0.74 rearrangements per million years.


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