Analysis of synaptonemal complexes in the amphidiploid of Lolium multiflorum × Festuca drymeja

Genome ◽  
1990 ◽  
Vol 33 (6) ◽  
pp. 903-907 ◽  
Author(s):  
Huw M. Thomas

Synaptonemal complexes in the amphidiploid Lolium multiflorum (4χ) × Festuca drymeja (4χ) have been examined by the whole mount spreading technique in nuclei with between 49 and 100% pairing. At mid to late zygotene nonhomologous associations are formed, with multivalents involving more than half the total axial element length in some cases. However, they are corrected by pachytene. There is evidence of differences in the timing or rate of pairing between the two sets of chromosomes; the L. multiflorum chromosomes seem more advanced than the F. drymeja chromosomes in their pairing at mid and late zygotene, and it is possible that this asynchrony places a constraint on intergenomic chromosome pairing.Key words: Lolium-Festuca, amphidiploid, synaptonemal complexes, nonhomologous pairing, correction mechanism.

Genome ◽  
1994 ◽  
Vol 37 (2) ◽  
pp. 181-189 ◽  
Author(s):  
Huw M. Thomas ◽  
Barry J. Thomas

A spreading technique for synaptonemal complexes (SCs) was applied to pollen mother cells of two aneuploid genotypes of autotriploid Lolium multiflorum (2n = 3x + 1 = 22). In the earliest nuclei analyzed the axial elements are in six groups of 3 and one group of 4. Most groups have formed multivalents with from one to five pairing partner exchanges, but there are also groups that have formed bivalents and univalents. Some axial elements have formed triple associations, in one case for the length of the trivalent. Unsynapsed axial elements remain aligned with their homologous SCs into pachytene, but this alignment is abolished as these axes pair heterologously among themselves until the entire axial element complement is synapsed. At metaphase I most chromosomes are associated as trivalents and quadrivalents.Key words: Lolium, triploid, pairing partner exchange, chiasma, multivalent.


Genome ◽  
1990 ◽  
Vol 33 (4) ◽  
pp. 465-471 ◽  
Author(s):  
Hum M. Thomas ◽  
W. G. Morgan

The synaptonemal complexes in the diploid hybrid Lolium multiflorum × Festuca drymeja were examined by the surface spreading technique, and chromosome pairing at metaphase I was analysed. Synaptonemal complex analysis revealed "illegitimate" pairing, including multivalents and foldback pairing. At metaphase I, most chiasmata were between chromosomes of the same genome, and again multivalents were found. It was concluded that most synaptonemal complexes resulted in chiasma formation. The effects of the large differences in DNA values of the two species and the possible genotypic effect of F. drymeja on chromosome pairing are discussed.Key words: Lolium-Festuca, synaptonemal complexes, nonhomologous pairing, DNA values.


2003 ◽  
Vol 160 (5) ◽  
pp. 657-670 ◽  
Author(s):  
Maureen Eijpe ◽  
Hildo Offenberg ◽  
Rolf Jessberger ◽  
Ekaterina Revenkova ◽  
Christa Heyting

In meiotic prophase, the sister chromatids of each chromosome develop a common axial element (AE) that is integrated into the synaptonemal complex (SC). We analyzed the incorporation of sister chromatid cohesion proteins (cohesins) and other AE components into AEs. Meiotic cohesin REC8 appeared shortly before premeiotic S phase in the nucleus and formed AE-like structures (REC8-AEs) from premeiotic S phase on. Subsequently, meiotic cohesin SMC1β, cohesin SMC3, and AE proteins SCP2 and SCP3 formed dots along REC8-AEs, which extended and fused until they lined REC8-AEs along their length. In metaphase I, SMC1β, SMC3, SCP2, and SCP3 disappeared from the chromosome arms and accumulated around the centromeres, where they stayed until anaphase II. In striking contrast, REC8 persisted along the chromosome arms until anaphase I and near the centromeres until anaphase II. We propose that REC8 provides a basis for AE formation and that the first steps in AE assembly do not require SMC1β, SMC3, SCP2, and SCP3. Furthermore, SMC1β, SMC3, SCP2, and SCP3 cannot provide arm cohesion during metaphase I. We propose that REC8 then provides cohesion. RAD51 and/or DMC1 coimmunoprecipitates with REC8, suggesting that REC8 may also provide a basis for assembly of recombination complexes.


Author(s):  
Kit W. Lee

The structure of the polycomplexes has been extensively studied in recent years (1,2,3). Theres structures are usually described as stacks or aggregates of the tripartite synaptonemal complexes. Although the presence of the synaptonemal complexes is a consistent feature in the late zygotene or pachytene stages of chlasmate meiosis and Its role in the processes of chromosome synapsis and crossing over has been suggested (4), the function of the polycomplexes remains obscure. Most of our understandings of the polycomplexes are obtained from the observations during the gametogenesis of the insects, and only a few examples of this structure in fungi and higher plants have been reported. The present study examines the occurrence of polycomplexes during the sporogenesis in the primitive vascular plant. Sporangia at different developmental stages were fixed with 3% glutaraldehyde in 0.1 M phosphate buffer, and postflxed in 2% osmium tetroxide. Dehydration was carried out with the ethanol series followed by embedding in Epon 812. Ultrathln sections were stained with uranyl acetate and lead citrate.


1977 ◽  
Vol 25 (1) ◽  
pp. 1-15
Author(s):  
L.L. Tres

Autosomes and sex chromosomes of mouse spermatocytes were examined during zygotene, pachytene, and diplotene by a whole-mount electron-microscope technique after cell dispersion in a detergent solution (Nonidet-P40). Zygotene, pachytene, and diplotene stages can be adequately identified in the preparations. Thus, asynchronous side-by-side pairing of homologous autosomes, some of them displaying attached nucleoli, defines zygotene. Pachytene is identified by complete pairing of homologues. Diplotene is characterized by disjunction of bivalents (autosomes and sex chromosomes), lack of autosomal-attached nucleoli, divergent expansions observed at lateral element endings of disassembled synaptonemal complexes, end-to-end association of the XY pair and well defined outward deformations (‘bulges’) along sex chromosomal axial cores. X and Y chromosomes display at pachytene an extensive side-by-side pairing segment which decreases in length as meiotic prophase advances. Each sex chromosomal axial core appears double and is formed by close apposition of 2 nearly parallel elements displayed separately along the entire length of the chromosomal core. This double structural feature suggests that each sex chromosomal axial core is presumably composed of 2 chromatid axial cores, each of which, in turn, constitutes the respective lateral elements of short synaptonemal complexes observed at the unpaired segment.


Genome ◽  
1988 ◽  
Vol 30 (1) ◽  
pp. 44-47 ◽  
Author(s):  
David W. Hale ◽  
Ira F. Greenbaum

The pattern of chromosomal pairing was analyzed in male deer mice (Peromyscus maniculatus and Peromyscus sitkensis) heterozygous for the presence of heterochromatic short arms. G- and C-banding of somatic metaphases indicated that the presence of heterochromatic short arms increased the length of chromosome 4 by 15% in P. sitkensis and that of chromosome 8 by 9% in P. maniculatus. Analysis of silver-stained late zygotene and early pachytene nuclei revealed a low frequency of unequal axial lengths in the synaptonemal complexes corresponding to the heteromorphic bivalents. All mid- and late pachytene nuclei, however, exhibited fully paired synaptonemal complexes with equalized axial lengths. These observations suggest the existence of an adjustment mechanism which functions to equalize the lengths of the two axes of the heteromorphic synaptonemal complex.Key words: synaptonemal complex, Peromyscus, heterochromatin, chromosomal polymorphism, synaptic adjustment.


Author(s):  
E. T. O'Toole ◽  
R. R. Hantgan ◽  
J. C. Lewis

Thrombocytes (TC), the avian equivalent of blood platelets, support hemostasis by aggregating at sites of injury. Studies in our lab suggested that fibrinogen (fib) is a requisite cofactor for TC aggregation but operates by an undefined mechanism. To study the interaction of fib with TC and to identify fib receptors on cells, fib was purified from pigeon plasma, conjugated to colloidal gold and used both to facilitate aggregation and as a receptor probe. Described is the application of computer assisted reconstruction and stereo whole mount microscopy to visualize the 3-D organization of fib receptors at sites of cell contact in TC aggregates and on adherent cells.Pigeon TC were obtained from citrated whole blood by differential centrifugation, washed with Ca++ free Hank's balanced salts containing 0.3% EDTA (pH 6.5) and resuspended in Ca++ free Hank's. Pigeon fib was isolated by precipitation with PEG-1000 and the purity assessed by SDS-PAGE. Fib was conjugated to 25nm colloidal gold by vortexing and the conjugates used as the ligand to identify fib receptors.


Author(s):  
T. L. Hayes

Biomedical applications of the scanning electron microscope (SEM) have increased in number quite rapidly over the last several years. Studies have been made of cells, whole mount tissue, sectioned tissue, particles, human chromosomes, microorganisms, dental enamel and skeletal material. Many of the advantages of using this instrument for such investigations come from its ability to produce images that are high in information content. Information about the chemical make-up of the specimen, its electrical properties and its three dimensional architecture all may be represented in such images. Since the biological system is distinctive in its chemistry and often spatially scaled to the resolving power of the SEM, these images are particularly useful in biomedical research.In any form of microscopy there are two parameters that together determine the usefulness of the image. One parameter is the size of the volume being studied or resolving power of the instrument and the other is the amount of information about this volume that is displayed in the image. Both parameters are important in describing the performance of a microscope. The light microscope image, for example, is rich in information content (chemical, spatial, living specimen, etc.) but is very limited in resolving power.


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.


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