Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 27
Filter
Add more filters










Publication year range
1.
Cytogenet Genome Res ; 142(1): 54-8, 2014.
Article in English | MEDLINE | ID: mdl-24217371

ABSTRACT

We analyzed the distribution of 2 repetitive DNAs, i.e. ribosomal DNA (rDNA) and a satellite DNA (satDNA), on the B chromosomes found in 17 natural populations of the grasshopper Eyprepocnemis ploransplorans sampled around the western Mediterranean region, including the Iberian Peninsula, Balearic Islands, Sicily, and Tunisia. Based on the amount of these repetitive DNAs, 4 types of B variants were found: B1, showing an equal or higher amount of rDNA than satDNA, and 3 other variants, B2, B24 and B5, bearing a higher amount of satDNA than rDNA. The variants B1 and B2 varied in size among populations: B1 was about half the size of the X chromosome in Balearic Islands, but two-thirds of the X in Iberian populations at Alicante, Murcia and Albacete provinces. Likewise, B2 was about one-third the size of the X chromosome in populations from the Granada province but half the size of the X in the populations collected at Málaga province. The widespread geographical distribution of the B1 variant makes it the best candidate for being the ancestor B chromosome in the whole western Mediterranean region.


Subject(s)
Biological Evolution , Chromosomes, Insect/ultrastructure , Grasshoppers/genetics , Animals , DNA, Ribosomal/genetics , DNA, Satellite/genetics , Evolution, Molecular , In Situ Hybridization, Fluorescence , Male , Mediterranean Region , Phylogeography , Species Specificity
2.
Chromosoma ; 122(1-2): 77-91, 2013 Mar.
Article in English | MEDLINE | ID: mdl-23283389

ABSTRACT

The cohesin complex plays a key role for the maintenance of sister chromatid cohesion and faithful chromosome segregation in both mitosis and meiosis. This complex is formed by two structural maintenance of chromosomes protein family (SMC) subunits and two non-SMC subunits: an α-kleisin subunit SCC1/RAD21/REC8 and an SCC3-like protein. Several studies carried out in different species have revealed that the distribution of the cohesin subunits along the chromosomes during meiotic prophase I is not regular and that some subunits are distinctly incorporated at different cell stages. However, the accurate distribution of the different cohesin subunits in condensed meiotic chromosomes is still controversial. Here, we describe the dynamics of the cohesin subunits SMC1α, SMC3, RAD21 and SA1 during both meiotic divisions in grasshoppers. Although these subunits show a similar patched labelling at the interchromatid domain of metaphase I bivalents, SMCs and non-SMCs subunits do not always colocalise. Indeed, SA1 is the only cohesin subunit accumulated at the centromeric region of all metaphase I chromosomes. Additionally, non-SMC subunits do not appear at the interchromatid domain in either single X or B chromosomes. These data suggest the existence of several cohesin complexes during metaphase I. The cohesin subunits analysed are released from chromosomes at the beginning of anaphase I, with the exception of SA1 which can be detected at the centromeres until telophase II. These observations indicate that the cohesin components may be differentially loaded and released from meiotic chromosomes during the first and second meiotic divisions. The roles of these cohesin complexes for the maintenance of chromosome structure and their involvement in homologous segregation at first meiotic division are proposed and discussed.


Subject(s)
Cell Cycle Proteins/genetics , Chromosomal Proteins, Non-Histone/genetics , Chromosomes/genetics , Grasshoppers , Meiosis/genetics , Anaphase/genetics , Animals , Centromere/genetics , Chromosome Segregation/genetics , Grasshoppers/cytology , Grasshoppers/genetics , Meiotic Prophase I/genetics , Mitosis/genetics , Cohesins
3.
Cytogenet Genome Res ; 128(1-3): 139-51, 2010.
Article in English | MEDLINE | ID: mdl-20389032

ABSTRACT

In the present study, and as a sincere tribute from the Cytogenetics teams from Madrid to Professor Máximo Drets on his 80th birthday, we have analyzed and compared 3 different grasshopper species with different synaptic patterns, a standard pattern, a second pattern with synapsis restricted to the proximal regions, and a third pattern with synapsis restricted to the distal regions. In the 3 species we have thoroughly analyzed the relationships among cohesin axis morphogenesis, formation of double strand breaks (DSBs) and recombination initiation. Our results demonstrate that in every case recombination initiation precedes synapsis, and that there is a direct relationship between the absence of meiotic recombination and the existence of particular unsynapsed chromosomal regions during prophase I. Based on our results we propose and discuss the mechanisms underlying the existence of incomplete synapsis and the localization of chiasma in wild species.


Subject(s)
Grasshoppers/genetics , Animals , Chromosome Pairing , Chromosomes , Crossing Over, Genetic , Grasshoppers/classification , Male , Recombination, Genetic
4.
Genome Dyn ; 5: 137-156, 2009.
Article in English | MEDLINE | ID: mdl-18948713

ABSTRACT

Most of the meiotic literature is based on species with monocentric chromosomes, however meiosis in protoctist, plant and animal species with holocentric chromosomes is less characterized. In some cases, an inverted meiotic sequence is claimed to occur, in which segregation of homologs is postponed until the second meiotic division. Additionally, other features also deserve interest, namely: (i) the different behavior of sex chromosomes if compared to that of the autosomes; (ii) the absence of a canonical kinetochore structure; (iii) the restriction of the kinetic activity to the chromosomal ends; (iv) the variations in the orientation of bivalents at the division plate, and (v) the possible occurrence of chiasma terminalization. Here we summarize the current knowledge on these topics in the meiosis of Hemiptera (Heteroptera) and present novel results that illustrate some of the special features mentioned above. We also point out the necessity of reviewing the term 'inverted meiosis' and propose some future prospects to study this peculiar meiosis.


Subject(s)
Hemiptera/cytology , Meiosis , Models, Genetic , Animals , Chromosomes/genetics , Crossing Over, Genetic , Kinetics
5.
Cytogenet Genome Res ; 106(2-4): 302-8, 2004.
Article in English | MEDLINE | ID: mdl-15292607

ABSTRACT

We have analysed the chromosome organisation and the location and temporal appearance of different proteins in X and B chromosomes in the grasshopper Eyprepocnemis plorans throughout the first meiotic prophase. We have used adult males that carry a B chromosome collected in natural Spanish populations. The scaffold organisation has been analysed by means of silver stained chromatid cores. In addition, we have detected by immunolabelling the presence of phosphoepitopes, the ensemble of cohesin axes, the location of histone gamma-H2AX, and recombinase Rad51. Our observations demonstrate that X and B chromosomes share similarities in chromatin organisation and in the expression of the tested proteins, which strongly differ from those of the autosomes. These results could be interpreted either as a support to the hypothesis that the Bs analysed here originated from the X chromosome, and/or that their chromatin composition and precocious condensation could determine their meiotic behaviour.


Subject(s)
Chromosomes/genetics , Grasshoppers/genetics , Meiosis/genetics , X Chromosome/genetics , Animals , Antibodies, Phospho-Specific/metabolism , DNA-Binding Proteins/genetics , Histones/immunology , Histones/metabolism , Male , Metaphase/genetics , Phosphorylation , Rad51 Recombinase
6.
Cytogenet Genome Res ; 98(2-3): 221-4, 2002.
Article in English | MEDLINE | ID: mdl-12698008

ABSTRACT

Heterogeneity for the length of telomeric DNA sequences has been found among different mitotic chromosomes in several mammalian species. However, there are no studies reporting such heterogeneity in meiotic chromosomes. To analyse this heterogeneity we have performed fluorescence in situ hybridization with a telomeric (C(3)TA(2))(3) peptide nucleic acid (PNA) probe on spread metaphase chromosomes during both male mouse meiotic divisions. Our results show that independently of the meiotic division, telomeric DNA signals were always surrounded by DAPI-stained chromatin, even at centromeric regions. Moreover, we have found heterogeneity for the size of telomeric DNA signals among different chromosomes, between homologues, and even within a given chromosome. We discuss the functional significance of the location of telomeric DNA in condensed meiotic chromosomes, and then the possible origin for the different polymorphisms found.


Subject(s)
Meiosis , Polymorphism, Genetic , Telomere/genetics , Animals , Chromosomes, Mammalian , DNA/genetics , In Situ Hybridization, Fluorescence , Male , Metaphase , Mice , Mice, Inbred C57BL , Telomere/chemistry
7.
Nat Cell Biol ; 3(8): 761-6, 2001 Aug.
Article in English | MEDLINE | ID: mdl-11483963

ABSTRACT

Cohesins, which have been characterized in budding yeast and Xenopus, are multisubunit protein complexes involved in sister chromatid cohesion. Regulation of the interactions among different cohesin subunits and the assembly/disassembly of the cohesin complex to chromatin are key steps in chromosome segregation. We previously characterized the mammalian STAG3 protein as a component of the synaptonemal complex that is specifically expressed in germinal cells, although its function in meiosis remains unknown. Here we show that STAG3 has a role in sister chromatid arm cohesion during mammalian meiosis I. Immunofluorescence results in prophase I cells suggest that STAG3 is a component of the axial/lateral element of the synaptonemal complex. In metaphase I, STAG3 is located at the interchromatid domain and is absent from the chiasma region. In late anaphase I and the later stages of meiosis, STAG3 is not detected. STAG3 interacts with the structural maintenance chromosome proteins SMC1 and SMC3, which have been reported to be subunits of the mitotic cohesin complex. We propose that STAG3 is a sister chromatid arm cohesin that is specific to mammalian meiosis I.


Subject(s)
Chromatids/genetics , Chromosome Segregation/physiology , Mammals/genetics , Meiosis/genetics , Nuclear Proteins/genetics , Sister Chromatid Exchange/genetics , Animals , Cell Cycle Proteins , Centromere/genetics , Centromere/metabolism , Chromosomal Proteins, Non-Histone , DNA-Binding Proteins , Fluorescent Antibody Technique , Fungal Proteins , Haplorhini , Male , Mammals/metabolism , Mice , Nuclear Proteins/metabolism , Organ Culture Techniques , Spermatocytes/cytology , Spermatocytes/metabolism , Testis/cytology , Testis/metabolism , Cohesins
8.
Chromosoma ; 110(7): 478-86, 2001 Dec.
Article in English | MEDLINE | ID: mdl-11862455

ABSTRACT

In somatic cells colchicine promotes the arrest of cell division at prometaphase, and chromosomes show a sequential loss of sister chromatid arm and centromere cohesion. In this study we used colchicine to analyse possible changes in chromosome structure and sister chromatid cohesion in prometaphase I-arrested bivalents of the katydid Pycnogaster cucullata. After silver staining we observed that in colchicine-arrested prometaphase I bivalents, and in contrast to what was found in control bivalents, sister kinetochores appeared individualised and sister chromatid axes were completely separated all along their length. However, this change in chromosome structure occurred without loss of sister chromatid arm cohesion. We also employed the MPM-2 monoclonal antibody against mitotic phosphoproteins on control and colchicine-treated spermatocytes. In control metaphase I bivalents this antibody labelled the tightly associated sister kinetochores and the interchromatid domain. By contrast, in colchicine-treated prometaphase I bivalents individualised sister kinetochores appeared labelled, but the interchromatid domain did not show labelling. These results support the notion that MPM-2 phosphoproteins, probably DNA topoisomerase IIalpha, located in the interchromatid domain act as "chromosomal staples" associating sister chromatid axes in metaphase I bivalents. The disappearance of these chromosomal staples would induce a change in chromosome structure, as reflected by the separation of sister kinetochores and sister axes, but without a concomitant loss of sister chromatid cohesion.


Subject(s)
Chromatids/physiology , Chromosomes/physiology , Colchicine/pharmacology , Grasshoppers/genetics , Sister Chromatid Exchange/drug effects , Animals , Chromatids/drug effects , Chromosomes/drug effects , Kinetochores/drug effects , Kinetochores/physiology , Male , Meiosis/drug effects , Meiosis/genetics , Metaphase , Microscopy, Fluorescence , Spermatozoa/cytology , Spermatozoa/ultrastructure , X Chromosome/ultrastructure
9.
Chromosoma ; 109(1-2): 35-43, 2000.
Article in English | MEDLINE | ID: mdl-10855493

ABSTRACT

It has been suggested that in species with monocentric chromosomes axial element (AE) components may be responsible for sister chromatid cohesion during meiosis. To test this hypothesis in species with holocentric chromosomes we selected three heteropteran species with different sex-determining mechanisms. We observed in surface-spreads and sections using transmission electron microscopy that the univalent sex chromosomes form neither AEs nor synaptonemal complexes (SCs) during pachytene. We also found that a polyclonal antibody recognizing SCP3/Cor1, a protein present at AEs and SC lateral elements of rodents, labels the autosomal SCs but not AEs or SC stretches corresponding to the sex chromosomes. Cytological analysis of the segregational behaviour of the sex univalents demonstrates that although these chromosomes segregate equationally during anaphase I they never show precocious separation of sister chromatids during late prophase I or metaphase I. These results suggest that AEs are not responsible for sister cohesion in sex chromosomes. The segregational behaviour of these chromosomes during both meiotic divisions also indicates that different achiasmate modes of chromosome association exist in heteropteran species.


Subject(s)
Heteroptera/genetics , Meiosis/genetics , Sex Chromosomes/genetics , Sister Chromatid Exchange/genetics , Animals , Cell Division/genetics , Cell Nucleus/ultrastructure , Chromatids/genetics , Chromatids/metabolism , Chromatids/ultrastructure , Chromosome Segregation/genetics , Karyotyping , Male , Sex Chromosomes/ultrastructure , Sister Chromatid Exchange/physiology
10.
Chromosome Res ; 8(1): 17-25, 2000.
Article in English | MEDLINE | ID: mdl-10730585

ABSTRACT

The meiotic behaviour of the X chromosome and one autosomal pair of the heteropteran Triatoma infestans was analysed by means of C-banding plus DAPI staining. At first metaphase, the X univalent is oriented with its long axis parallel to the equatorial plate, which suggests a holocentric interaction with the spindle fibres. After this initial orientation, kinetic activity is restricted to one of both chromatid ends. The election of the active chromatid end is random and it is independent of the end selected in the sister chromatid. At second metaphase, the X and Y chromatids associate side by side forming a pseudobivalent. After that, the kinetic activity is again restricted to either of both chromosomal ends in a random fashion. At first metaphase, the fourth autosomal bivalent shows two alternative random orientations depending on the chromosome end showing kinetic activity (DAPI positive or opposite). At second metaphase, half bivalents are oriented with their long axis parallel to the equatorial plate. Three different segregation patterns are observed. The kinetic activity can be localised: (i) in the end with the DAPI signal (46.9%), (ii) in the opposite end (44.6%) or (iii) in one DAPI-positive end in one chromatid and in the opposite end in the other one (8.5%). The existence of the last pattern indicates that the same end can show kinetic activity during both meiotic divisions. Our results provide new information on the comparative meiotic behaviour of autosomes and sex chromosomes in holocentric systems.


Subject(s)
Chromosomes , Meiosis , Sex Chromosomes , Triatoma/genetics , Animals , Azure Stains , Chromosome Banding , Fluorescent Dyes , Indoles
11.
J Cell Sci ; 112 ( Pt 17): 2957-69, 1999 Sep.
Article in English | MEDLINE | ID: mdl-10444390

ABSTRACT

Sister-chromatid arm cohesion is lost during the metaphase I/anaphase I transition to allow homologue separation. To obtain needed information on this process we have analysed in grasshopper bivalents the sequential release of arm cohesion in relation to the behaviour of chromatid axes. Results show that sister axes are associated during early metaphase I but separate during late metaphase I leading to a concomitant change of chromosome structure that implies the loss of sister-kinetochore cohesion. Afterwards, homologues initiate their separation asynchronously depending on their size, and number and position of chiasmata. In all bivalents thin chromatin strands at the telomeres appeared as the last point of contact between sister chromatids. Additionally, we have analysed the participation of phosphoproteins recognised by the MPM-2 monoclonal antibody against mitotic phosphoproteins in arm cohesion in bivalents and two different kinds of univalents. Results show the absence of MPM-2 phosphoproteins at the interchromatid domain in mitotic chromosomes and meiotic univalents, but their presence in metaphase I bivalents. These phosphoproteins are lost at the onset of anaphase I. Taken together, these data have prompted us to propose a 'working' model for the release of arm cohesion during meiosis I. The model suggests that MPM-2 phosphoproteins may act as cohesive proteins associating sister axes. Their modification, once all bivalents are correctly aligned at the metaphase plate, would trigger a change of chromosome structure and the sequential release of sister-kinetochore, arm, and telomere cohesions.


Subject(s)
Insect Proteins/physiology , Meiosis/physiology , Phosphoproteins/physiology , Sister Chromatid Exchange/physiology , Spermatocytes/ultrastructure , Anaphase , Animals , Chromatids/ultrastructure , Grasshoppers , Kinetochores/physiology , Male , Metaphase , Microscopy, Electron , Microscopy, Fluorescence , Phosphorylation , Protein Processing, Post-Translational , Protein Serine-Threonine Kinases/metabolism , Silver Staining , Telomere/physiology , Telomere/ultrastructure
12.
Chromosome Res ; 6(8): 639-42, 1998 Dec.
Article in English | MEDLINE | ID: mdl-10099877

ABSTRACT

Several techniques have been developed for protein immunolocalization in meiotic cells. However, most of them include treatments that lead to cell disruption and are only suitable for prophase-I cells. We describe a novel squash procedure of cell preparation for protein immunolabelling of different meiotic stages. This procedure is an alternative to both cryosectioning and whole spreading procedures. We present results obtained in mouse spermatocytes with three different antibodies: the MPM-2 mAb against mitotic phosphoepitopes, an anticentromere serum and a polyclonal serum against the SCP3 protein of the axial elements and lateral elements of the synaptonemal complex. The procedure was tested for single and double immunolabelling. With this technique a large number of cells at different meiotic stages can be analysed. Cell stages are easily identified and cell and chromosome structures are preserved. Thus, it allows the study of chromosome behaviour and the relationships between the different structural elements of the cell throughout meiotic divisions. Our procedure is also suitable for three-dimensional (3D) analyses and proved to be reliable in a wide range of systems including insects and mammals. In addition, the procedure may be interesting to obtain a rapid immunological diagnosis.


Subject(s)
Centromere/chemistry , Immunohistochemistry/methods , Meiosis , Nuclear Proteins/analysis , Spermatocytes/chemistry , Animals , Antibodies, Monoclonal/immunology , Cell Cycle Proteins , Centromere/ultrastructure , Chromosomes/chemistry , DNA-Binding Proteins , Male , Mice , Mice, Inbred C57BL , Spermatocytes/cytology
13.
Chromosome Res ; 5(1): 47-56, 1997 Feb.
Article in English | MEDLINE | ID: mdl-9088643

ABSTRACT

The meiotic behaviour of the holocentric chromosomes of the heteropteran species Triatoma infestans has been analysed by means of orcein staining and C-banding on squashed spermatocytes. We have focused our analysis on chromosome 3, which shows a large distal heterochromatic band at one of the ends of both homologues. At metaphase I, and independently of the chiasma position, two alternative orientations have been observed: either the heterochromatic or the euchromatic ends of both homologues are directed to opposite poles. At anaphase I, the kinetic activity is restricted to the same chromosome end (euchromatic or heterochromatic) of each homologue. The frequencies of these two alternatives are not random and differ significantly among the five individuals analysed. However, the euchromatic ends present kinetic activity at a higher frequency than the heterochromatic ends. At metaphase II, half-bivalents also show the kinetic activity restricted to either of the chromosome ends (euchromatic or heterochromatic). The frequencies of each alternative are inverted in anaphase II compared with those scored in anaphase I. Accordingly, those ends that present kinetic activity at anaphase I segregate reductionally during the first meiotic division and equationally during the second meiotic division. These results provide sound evidence on the meiotic behaviour of holocentric chromosomes, as regards the absence of chiasma terminalization and the modes of orientation and segregation.


Subject(s)
Anaphase , Chromatids , Chromosomes/ultrastructure , Meiosis/genetics , Triatoma/genetics , Anaphase/physiology , Animals , Chromatids/genetics , Chromatin/genetics , Chromatin/ultrastructure , Chromosome Banding , Chromosomes/genetics , Euchromatin , Heterochromatin/genetics , Heterochromatin/ultrastructure , Meiosis/physiology , Metaphase/physiology , Prophase/physiology , Telomere/ultrastructure , X Chromosome , Y Chromosome
14.
J Histochem Cytochem ; 44(11): 1279-88, 1996 Nov.
Article in English | MEDLINE | ID: mdl-8918903

ABSTRACT

We analyzed first-metaphase meiotic chromosomes of the grasshopper Chorthippus jucundus by two different methods, i.e., a silver impregnation technique and the osmium tetroxide-p-phenylenediamine (Os-PPD) procedure. The former was applied on squashed testes previously fixed in ethanol-acetic acid, whereas for Os-PPD the material was not subjected to any previous extraction treatment but was fixed in OsO4, treated with PPD, and embedded in Epon 812. Both techniques revealed chromatid cores and kinetochores regardless of the processing of the material (squashed or sectioned). Unstained Os-PPD sections were analyzed by light microscopy and transmission electron microscopy (TEM). The Os-PPD technique provided a high contrast of chromatid cores and kinetochores in relation to the chromatin, which revealed a low electron density. To determine the Os-PPD reaction mechanism, the PAS procedure, as well as scanning electron microscopy (SEM) backscattering and SEM X-ray microanalysis, was performed on sections. By use of the Os-PPD-PAS procedure, glycol groups formed by oxidation of osmium bound to aromatic substrates were detected in chromatid cores and kinetochores by brightfield and fluorescence microscopy. A high Z contrast was detected in these structures by backscattered electron imaging. SEM X-ray microanalysis showed osmium and phosphorus to be the main elements present on the chromatid cores. Taking into account the known reactivity of OsO4 and the present results, the possible participation of nucleic acids as well as proteins in the Os-PPD reaction mechanism and in the composition of chromatid cores and kinetochores is discussed.


Subject(s)
Chromatids/ultrastructure , Chromosomes/ultrastructure , Kinetochores/ultrastructure , Meiosis , Animals , Grasshoppers , Histocytochemistry , Male , Microscopy, Electron , Osmium Tetroxide , Phenylenediamines , Staining and Labeling/methods
15.
Eur J Cell Biol ; 70(4): 352-60, 1996 Aug.
Article in English | MEDLINE | ID: mdl-8864664

ABSTRACT

In the present paper we have analyzed the trilaminar kinetochores present in the mitotic holocentric chromosomes of the heteropteran Graphosoma italicum by means of different cytochemical and immunocytochemical techniques at the ultrastructural level. The results obtained after EDTA and osmium tetroxide/p-phenylenediamine, both cytochemical methods for preferential detection of ribonucleoproteins, show that the inner and outer kinetochore layers are selectively contrasted. These results agree with those previously reported for the kinetochores of monocentric chromosomes. Additionally, using cytochemical techniques such as ethanolic phosphotungstic acid staining and silver impregnation, which recognize proteins, we report a different response of the inner kinetochore layer compared with those obtained in the middle and outer layers. Finally, our immunoelectron microscopic results employing monoclonal antibodies against DNA indicate the absence of this component from the kinetochore plates in holocentric chromosomes. Although the trilaminar appearance of kinetochores in holocentric chromosomes is very similar to that of the localized kinetochores in mammals, our observations imply the existence of significant differences on the macromolecular organization of both types of kinetochores.


Subject(s)
Chromosomal Proteins, Non-Histone/ultrastructure , Chromosomes/ultrastructure , Kinetochores/ultrastructure , Spermatogonia/ultrastructure , Animals , Antibodies, Monoclonal , Chromosomes/immunology , Hemiptera , Histocytochemistry , Immunohistochemistry , Male , Microscopy, Electron , Staining and Labeling/methods , Tissue Fixation/methods
16.
Chromosome Res ; 4(2): 124-32, 1996 Feb.
Article in English | MEDLINE | ID: mdl-8785606

ABSTRACT

We have determined the number and location of the nucleolar organizing regions in spermatocytes of Graphosoma italicum (2n = 12A + XYmale/XXfemale) by means of silver impregnation, chromomycin A3/distamycin A staining and fluorescence in situ hybridization. The identification of only one nucleolar organizing region located at one of the X chromosome ends has provided a suitable cytological marker to analyse the segregation of this univalent and that of the XY pseudobivalent during the first and second meiotic divisions respectively. Our results clearly show that at first meiotic metaphase the chromatids of the X chromosome are orientated with their long axes perpendicular to the polar axis. Although the kinetic activity is restricted to only one end in both X chromatids during the first meiotic division, both ends of the same chromatid have the same probability of showing such kinetic activity. In this sense, we also report that the chromatid segregation may be initiated either at the same sister chromatid ends or at opposite ends in each chromatid. Thus, this indicates a sex chromatid independence as regards to the chromatid segregation during the first meiotic division. Throughout the second meiotic division both ends of the X chromatid are involved with the same probability in the end-to-end association to conform the XY pseudobivalent. This also implies a random localization of the kinetic activity at the ends opposite to those involved in the end-to-end association.


Subject(s)
Chromatids/physiology , Insecta/physiology , Meiosis , Nucleolus Organizer Region , Spermatocytes/ultrastructure , X Chromosome/physiology , Animals , Chromatids/ultrastructure , Female , In Situ Hybridization, Fluorescence , Insecta/genetics , Male , Microscopy, Electron, Scanning , Sex Chromatin/ultrastructure , Silver Staining , Spermatocytes/physiology , X Chromosome/ultrastructure
17.
Chromosoma ; 104(4): 287-97, 1995 Dec.
Article in English | MEDLINE | ID: mdl-8565705

ABSTRACT

During early embryogenesis of the nematode Parascaris univalens (2n=2) the processes of chromatin diminution and segregation of the germ and somatic cell lineages take place simultaneously. In this study we analyzed the nucleolar cycle in early embryos, both in germinal and somatic blastomeres, by means of silver staining and antibodies against the nucleolar protein fibrillarin. We observed an identical nucleolar cycle in both types of blastomeres, hence, the chromatin diminution process has no effect on the nucleolar cycle of somatic blastomeres. We report the existence of outstanding differences between this cycle and those previously reported during early embryogenesis of other species. There is a true nucleolar cycle in early embryos that shows a peculiar nucleolar disorganization at prophase, and a preferential localization of prenucleolar bodies only on the euchromatic regions during nucleologenesis. Moreover, fibrillarin does not form a perichromosomal sheath in metaphase or anaphase holocentric chromosomes, probably owing to their special centromeric organization. The number and location of nucleolus organizer regions (NORs) in the chromosomal complement have been determined using silver impregnation, chromomycin A3/ distamycin A staining, and fluorescent in situ hybridization using an rDNA probe. There are only two NORs, one per chromosome, and these are located in the middle of the euchromatic central regions. This location implies that no rDNA sequences are lost in blastomeres after chromatin diminution. Moreover, the constant presence of two nucleoli in somatic blastomeres suggests that NORs are not affected during the fragmentation of euchromatic regions when this process occurs.


Subject(s)
Ascaridida/embryology , Cell Cycle , Cell Nucleolus/ultrastructure , Nucleolus Organizer Region/ultrastructure , Animals , Ascaridida/ultrastructure , Autoimmunity , Blastomeres/ultrastructure , Chromosomal Proteins, Non-Histone/immunology , Female , Horses/parasitology , In Situ Hybridization, Fluorescence , Staining and Labeling
18.
Chromosome Res ; 2(5): 361-8, 1994 Sep.
Article in English | MEDLINE | ID: mdl-7981940

ABSTRACT

We have analysed by means of silver staining the structure of the chromosome axis at the telomeres of meiotic chromosomes in three different grasshopper species. At metaphase I the chromatid axes run the length of the chromatids although they do not reach the chromosome ends. The axes of sister chromatids are associated and show a round differentiation at their distal ends that we have named the 'telochore'. Telochores never contact the chromosome ends: there is always some chromatin beyond them. In late metaphase I bivalents with a distal chiasma, anaphase I and metaphase II half-bivalents and anaphase II chromatids, the axes clearly possess one telochore in each chromosome end. These results seem to indicate that telochores are differentiations of the distal ends of chromatids. We discuss the possible structural significance of telochores according to the current scaffold/radial loop model of chromatin organization of eukaryotic metaphase chromosomes. Additionally, we suggest the possible functional role of the telochore as a nucleoprotein domain forming a protective cap for telomeric DNA.


Subject(s)
Grasshoppers/genetics , Telomere/ultrastructure , Animals , Base Sequence , Chromatids/ultrastructure , Male , Meiosis , Molecular Sequence Data , Nucleoproteins/analysis , Silver Staining , Spermatocytes/ultrastructure
19.
Chromosome Res ; 2(5): 369-75, 1994 Sep.
Article in English | MEDLINE | ID: mdl-7981941

ABSTRACT

We describe a simple silver impregnation method for the ultrastructural detection of kinetochores on meiotic chromosomes of the grasshopper Eyprepocnemis plorans. Testes were fixed with glutaraldehyde and silver-impregnated. After Epon 812 embedding, ultrathin cutting and counterstaining with uranyl acetate, sections were studied by transmission electron microscopy. The meiotic chromosomes showed differentially silver-impregnated 'ball and cup' kinetochores. Some pericentriolar material also showed silver deposits. These observations are discussed in the light of previous results obtained by light microscopy of silver-stained spermatocytes in which both kinetochores and pericentriolar material were also preferentially stained. These results suggest a role for acidic proteins in the composition of these structures.


Subject(s)
Grasshoppers/genetics , Kinetochores/ultrastructure , Silver Staining , Animals , Male , Meiosis , Spermatocytes/ultrastructure
20.
Chromosoma ; 102(7): 491-9, 1993 Jul.
Article in English | MEDLINE | ID: mdl-7690696

ABSTRACT

The two nucleolus organizing chromosome pairs of the grasshopper Pyrgomorpha conica can carry a proximal supernumerary heterochromatic segment. We employed different cytological techniques to characterize and analyze the possible origin of this segment. The supernumerary segment and the nucleolus organizing regions (NORs) show similar responses after C-banding plus either Giemsa or acridine orange, and chromomycin A3/distamycin A staining to detect GC-rich chromosome regions. Fluorescence in situ hybridization with a biotinylated rDNA probe demonstrated that the segment originated by amplification of the rDNA genes. However, as the silver staining indicates, the ribosomal genes present in the segment are not active since no nucleolus is formed. The use of in situ digestion with the isoschizomeric MspI and HpaII restriction endonucleases and subsequent Giemsa, ethidium bromide or chromomycin A3/distamycin A staining, suggests that the segment has been inactivated by DNA methylation.


Subject(s)
DNA, Ribosomal/genetics , Grasshoppers/genetics , Heterochromatin/ultrastructure , Animals , Chromosome Banding , DNA Restriction Enzymes , DNA, Ribosomal/chemistry , Grasshoppers/ultrastructure , In Situ Hybridization, Fluorescence , Karyotyping , Male , Methylation , Nucleolus Organizer Region/ultrastructure , Staining and Labeling
SELECTION OF CITATIONS
SEARCH DETAIL
...