Orientation and segregation of Robertsonian trivalents in Dichroplus pratensis (Acrididae)

Pairing behavior, metaphase I orientation, and anaphase I segregation of centric fusion trivalents were studied in 26 single, 15 double, and 2 triple male fusion heterozygotes of the polymorphic South American melanopline grasshopper Dichroplus pratensis. They represent the seven different fusions and their combinations already described in different populations of the species. Our analysis showed the following: (1) pairing behavior is very regular in all trivalents; (2) frequencies of linear orientation was very low irrespective of the trivalent involved; (3) reorientation seems to occur frequently since frequencies of abnormal segregation and aneuploid second division cells were invariably lower than those of nonconvergent orientation; (4) aneuploidy and abnormal sperm production increases with increasing number of fusions; (5) chiasma frequency and localisation is relevant to trivalent orientation since trivalents with nonconvergent orientations showed proximal and interstitial chiasmata more frequently than convergently oriented ones. The results are in agreement with the hypothesis that these polymorphisms are old and stable, and confirm that for the maintenance of a balanced polymorphism, if this polymorphism is adaptive because of its consequences on recombination, position effects, etc., changes tending to stabilise trivalent orientation and segregation are central.     

Effects of centric fusions on chiasma frequency and position in Leptysma argentina (Acrididae: Orthoptera) I. Spontaneous and stable polymorphic centric fusions

Leptysma argentina is a highly polymorphic South-American grasshopper from the cytological point of view; all populations so far studied carry a polymorphic fusion between pairs 3 and 6. In heterozygotes, the trivalent 3-3/6-6 shows alternate orientation in 97.17% of the cells. Trivalent chiasma frequency is significantly lower than in the combined 3 and 6 bivalents of the standard homozygote; besides, there is a marked displacement of chiasmata to a distal position. In structural homozygotes the same effects, but not so marked, were observed.One individual was a double heterozygote for both the polymorphic centric fusion and a spontaneous one between pairs 5 and 7. The presence of a fragment, sometimes associated with the centromeric region of the nonfused 5 chromosome, was detected in more than 50% of the cells. The orientation of trivalent 5-5/7-7 in metaphase I was highly irregular (36% linear orientation). Neither frequency nor position of chiasmata were altered in trivalent 5-5/7-7 when compared with bivalents 5+7 of normal individuals.The results suggest that proximal and interstitial chiasma reduction observed in trivalent 3-3/6-6 of L. argentina is due to a later adaptation to the polymorphic condition or a fortuitous genetic condition present in the original mutant, rather than to a direct effect of the fusion itself on chiasma distribution.Fellow of the Consejo Nacional de Investigaciones Cientificas y Técnicas (CONICET)     

Differential mechanisms governing segregation of a univalent in oocytes and spermatocytes of Drosophila melanogaster

In tricomplex heterozygotes in Drosophila melanogaster three metacentric autosomes (the TRI chromosomes) appear as a trivalent in meiosis while one autosome consisting of two homologous arms attached to the same centromere (a compound) behaves as an obligatory univalent. Cytological analysis of meiosis of tricomplex heterozygotes indicates that in oocytes the univalent compound behaves non-independently in relation to segregation of the trivalent. The compound is distributed preferentially to the same pole as one TRI chromosome. In spermatocytes the compound is distributed at random. In some oocytes the directed segregation is shown to be due to a disjunctional interaction between the compound and one partner of the trivalent at the same time as the other two chromosomes of the trivalent are separating from each other. The basic difference between the segregational mechanisms in the two sexes is discussed with a review of evidence indicating that in males segregation is determined by physical linkage that produces a stable orientation of the homologues at metaphase I. On the other hand, both genetic and cytological evidence indicate that in females a physical linkage (a chiasma) is non-essential for maintenance of co-orientation and stability after the onset of prometaphase. Genetic and cytological evidence support the hypothesis that disjunction is predetermined by non-random arrangement of the centromeric regions of chromosomes in the chromocentre — a suprachromosomal organization characteristic of maturing oocytes.by D. Schweizer     

M chromosome of the wild silkworm, Bombyx mandarina (n = 27), corresponds to two chromosomes in the domesticated silkworm, Bombyx mori (n = 28).

Chromosomes of Bombyx mori (n = 28) and of Bombyx mandarina (n = 27) were studied cytogenetically to resolve the origin of the large M chromosome in the Japaneses type of B. mandarina. In the F1 progeny from the reciprocal cross between B. mandarina and B. mori, the mitotic chromosome number was 2n = 55, and a chromosome configuration of 26 bivalents plus 1 trivalent was observed at metaphase I of germ cells. The trivalent chromosome consisted of the M chromosome from B. mandarina and two chromosomes from B. mori. When males of B. mori were mated to the F1 females, nuclei with two types of chromosome number (2n = 55 and 2n = 56) and two sets of chromosome pairs (26 bivalents plus 1 trivalent versus 28 bivalents) were observed in the metaphase I stage. Linkage analysis showed that the 14th chromosome of B. mori was involved in these two types of chromosome segregation. This result indicates that the M chromosome in B. mandarina arose from a fusion between a chromosome corresponding to the 14th linkage group and another, yet unidentified linkage group.     

Chromosomal synapsis and the meiotic process in male mesquite lizards, Sceloporus grammicus complex.

Meiosis in males of the F5 cytotype of Sceloporus grammicus was examined through the analysis of synaptonemal complexes (SCs), diakinetic (metaphase I) nuclei, and secondary spermatocytes (metaphase II configurations). These data allowed the establishment of criteria for substaging of zygonema and pachynema, morphological characterization of the SC complement, and comparison of the orientation and segregation of the autosomes and sex chromosomes. The analysis of nuclei from all stages of meiotic prophase I (leptonema through diakinesis) provided a useful means of partitioning the temporal sequence of early meiotic events. Three substages of zygonema (Z1-Z3) were established, based on the extent of synapsis of the microchromosomal and macrochromosomal elements. Synaptic initiation of the autosomes and sex chromosomes was synchronous. Two patterns of macrochromosomal synapsis were observed. Whereas synapsis of the biarmed elements was biterminal (i.e., progressing from both ends of the homologs), synapsis of the acrocentric elements was uniterminal involving only the distal (noncentromeric) ends of the homologs. Unique sex-chromosomal characteristics were not observed in S. grammicus and, therefore, the substaging of pachynema was based on subjective criteria. Examination of diakinesis--metaphase I and metaphase II configurations indicated low levels of diakinetic irregularities with balanced segregation of the autosomal bivalents and the sex-chromosomal trivalent.     

Male meiosis and gametogenesis in wild house mice (Mus musculus domesticus) from a chromosomal hybrid zone; a comparison between "simple" Robertsonian heterozygotes and homozygotes.

Wild male house mice Mus musculus domesticus were collected from the hybrid zone between the John o'Groats race (2n = 32) and the standard race (2n = 40) in northern Scotland. Meiosis in both homozygotes (2n = 32, 36, and 40) and single Robertsonian heterozygotes (2n = 33, 35, and 37) was found to be orderly. At prophase/metaphase I in heterozygotes, a trivalent was formed from the metacentric and two homologous acrocentrics. At pachytene, this trivalent usually had a single side arm at the position of the centromeres, as a result of nonhomologous pairing of the acrocentrics. This side arm persisted into diplotene. Generally only a single chiasma was formed between each acrocentric and the metacentric. Anaphase I nondisjunction frequencies were estimated as 1.5% for the homozygotes and 2.7% for the heterozygotes. The extent of germ cell death between the pachytene and round spermatid stages was 18% greater in heterozygotes than in homozygotes. Our results concur with previous studies which indicate that single Robertsonian heterozygotes in wild house mice have near-normal fertility.     

Meiotic chromosome segregation in human t(11;22)(q23;q11) carriers: a theoretical consideration

The t(11;22)(q23;q11) translocation is the most frequently encountered familial reciprocal translocation in humans. In the majority of reported cases ascertainment has been through the birth of a child with the chromosomal constitution 47,XX,+der(22) or 47,XY,+der(22), i.e., tertiary trisomy. Previous segregation analysis of familial cases showed a number of interesting features. Thus, euploid unbalanced genotypes resulting from adjacent segregation are absent in the progeny, and only tertiary trisomic offspring are recovered. To explain this unusual progeny output we present here a model for the meiotic behavior of this translocation in the carriers based on an analysis of cytogenetic data of progeny of carriers. This model predicts the formation of a chain trivalent with chromosome order 11-der(11)-22 during prophase I and its predominant alternate orientation at metaphase I.     

Telomere-independent homologue pairing and checkpoint escape of accessory ring chromosomes in male mouse meiosis

We analyzed transmission of a ring minichromosome (MC) through mouse spermatogenesis as a monosome and in the presence of a homologue. Mice, either monosomic or disomic for the MC, produced MC+ offspring. In the monosomic condition, most univalents underwent self-synapsis as indicated by STAG3, SCP3, and SCP1 deposition. Fluorescent in situ hybridization and three-dimensional fluorescence microscopy revealed that ring MCs did not participate in meiotic telomere clustering while MC homologues paired at the XY-body periphery. Self-synapsis of MC(s) and association with the XY-body likely allowed them to pass putative pachytene checkpoints. At metaphase I and II, MC kinetochores assembled MAD2 and BUBR1 spindle checkpoint proteins. Unaligned MCs triggered the spindle checkpoint leading to apoptosis of metaphase cells. Other MCs frequently associated with mouse pericentric heterochromatin, which may have allowed them to pass the spindle checkpoint. Our findings indicate a telomere-independent mechanism for pairing of mammalian MCs, illuminate escape routes to meiotic checkpoints, and give clues for genetic engineering of germ line-permissive chromosomal vectors.     

Synaptonemal complex components persist at centromeres and are required for homologous centromere pairing in mouse spermatocytes

Recent studies in simple model organisms have shown that centromere pairing is important for ensuring high-fidelity meiotic chromosome segregation. However, this process and the mechanisms regulating it in higher eukaryotes are unknown. Here we present the first detailed study of meiotic centromere pairing in mouse spermatogenesis and link it with key events of the G2/metaphase I transition. In mouse we observed no evidence of the persistent coupling of centromeres that has been observed in several model organisms. We do however find that telomeres associate in non-homologous pairs or small groups in B type spermatogonia and pre-leptotene spermatocytes, and this association is disrupted by deletion of the synaptonemal complex component SYCP3. Intriguingly, we found that, in mid prophase, chromosome synapsis is not initiated at centromeres, and centromeric regions are the last to pair in the zygotene-pachytene transition. In late prophase, we first identified the proteins that reside at paired centromeres. We found that components of the central and lateral element and transverse filaments of the synaptonemal complex are retained at paired centromeres after disassembly of the synaptonemal complex along diplotene chromosome arms. The absence of SYCP1 prevents centromere pairing in knockout mouse spermatocytes. The localization dynamics of SYCP1 and SYCP3 suggest that they play different roles in promoting homologous centromere pairing. SYCP1 remains only at paired centromeres coincident with the time at which some kinetochore proteins begin loading at centromeres, consistent with a role in assembly of meiosis-specific kinetochores. After removal of SYCP1 from centromeres, SYCP3 then accumulates at paired centromeres where it may promote bi-orientation of homologous centromeres. We propose that, in addition to their roles as synaptonemal complex components, SYCP1 and SYCP3 act at the centromeres to promote the establishment and/or maintenance of centromere pairing and, by doing so, improve the segregation fidelity of mammalian meiotic chromosomes.     

Metaphase I orientation of chain-forming interchange quadrivalents: a theoretical consideration

The orientation behavior of chain forming interchange quadrivalents at metaphase I was studied in three interchange heterozygotes of pearl millet [Pennisetum americanum (L.) Leeke] which involve chromosomes 1, 3, 6 and 7 in various combinations. Of these, two combinations predominantly produced rings and the third was a chain-forming type. The chain quadrivalents derived from the two ring-forming interchanges, as well as the chain quadrivalent generated by the third interchange, all showed one adjacent orientation at metaphase I (adjacent-1 or -2, depending upon the formation or failure of chiasmata and their positions in the different segments of the pachytene cross). Homologous centromere co-orientation leading to adjacent-1 and alternate-1 occurs following chiasma failure in the noncentric arms of the pachytene cross, and nonhomologous centromere co-orientation leading to adjacent-2 and alternate-2 occurs following chiasma failure in the centric arms of the pachytene cross. Thus, it has been proposed that, unlike in ring quadrivalents, a specific chain quadrivalent will have only homologous or nonhomologous centromere co-orientations at metaphase I.     

Cytological evidence for holocentric chromosomes of the silkworms,Bombyx mori and B. mandarina, (Bombycidae,Lepidoptera)

The nature of the centromere and the orientation in meiosis of silkworm chromosomes were investigated using the trivalent of the F₁ hybrid between the wild and domestic silkworm and X-ray-induced aberrant chromosomes as well as normal silkworm chromosomes. The results of the experiments were as follows: (1) Pro-metaphase chromosomes showed no distinct primary constriction even after treatment with hypotonic solution, (2) sister chromatids separated in parallel along the entire length of the chromosome at mitotic anaphase, (3) chiasmata underwent complete terminalization during diakinesis and thus chromosome dyads were always connected end-to-end by a terminal chiasma at metaphase I, (4) radiation-induced aberrant chromosomes were stably transmitted throughout a number of cell generations, and (5) although the homomorphic bivalents generally orientated axially at metaphase I and equatorially at metaphase II, this normal sequence tended to be inverted or modified in the X-ray-induced aberrant chromosomes and in the trivalent of the F₁ hybrid silkworms. These observations may be best interpreted by assuming the holocentric nature of silkworm chromosomes.     

The behaviour of kinetochore microtubules during meiosis in the fungusSaprolegnia

Summary InSaprolegnia, kinetochore microtubules persist throughout the mitotic nuclear cycle but, whilst present at leptotene, they disappear coincidently with the formation of synaptonemal complexes at pachytene and reform at metaphase I. In some other fungi chromosomal segregation is random in meiosis and non-random in mitosis. The attachment of chromosomes to persistent kinetochore microtubules in mitosis, but not meiosis, inSaprolegnia provides a plausible explanation for such behaviour. At metaphase I each bivalent is connected to the spindle by 2 laterally paired kinetochore microtubules whereas at metaphase II (as in mitosis) each univalent bears only one kinetochore microtubule, thus showing that all kinetochores are fully active at all stages of meiosis.     

Holocentric plant meiosis: first sisters,then homologues

Meiosis is a crucial process of sexual reproduction by forming haploid gametes from diploid precursor cells. It involves 2 subsequent divisions (meiosis I and meiosis II) after one initial round of DNA replication. Homologous monocentric chromosomes are separated during the first and sister chromatids during the second meiotic division. The faithful segregation of monocentric chromosomes is realized by mono-orientation of fused sister kinetochores at metaphase I and by bi-orientation of sister kinetochores at metaphase II. Conventionally this depends on a 2-step loss of cohesion, along chromosome arms during meiosis I and at sister centromeres during meiosis II.     

Meiotic behavior of Robertsonian heterozygotes in populations of Dichroplus pratensis (Acrididae) with different fusion frequencies

The meiotic behavior of Robertsonian heterozygotes of Dichroplus pratensis was analyzed in order to establish the nature of the fusion polymorphisms (stable or transient) that occur in the species. The range of fusion frequencies varies widely for each fusion studied and populations with extreme frequencies exist, which could indicate a tendency for the loss or fixation of a given rearrangement. Our results revealed that no significant correlation exists between orientation (convergent and non-convergent) in PMI and MI and fusion frequency, nor between aneuploidy and fusion frequency. Thus, orientation and segregation seem to be independent of the frequencies of the fusions, which also do not appear to affect severely the fertility of heterozygotes. Our data suggest that the polymorphisms are, at least, cytogenetically stable.Both authors are affiliated to the CONICET (Argentina)     

Chiasma distributions and chromosome segregation in male and female translocation heterozygous mice analysed using FISH

Two factors postulated to influence the meiotic behaviour of reciprocal translocations were investigated. Firstly, variation in the length of translocated and non-translocated segments was studied in male mice using four different rearrangements involving chromosomes 2 and 4. Secondly, sex-related effects were analysed through comparison of the meiotic behaviour of two translocations in male and female germ cells. In the first part of the study, primary and secondary spermatocytes of male mice carrying a translocation [T(2;4)1Ca, T(2;4)13H, T(2;4)1Sn, or T(2;4)1Go] were screened. Each rearrangement had different proportions of cells with ring and chain quadrivalents at metaphase I; the T(2;4)1Sn heterozygote also had a high rate (45%) of translocation bivalents. In general, the translocations had elevated chiasma frequencies in the rearranged chromosomes compared with structurally normal chromosomes 2 and 4, although the extent of the effect varied. Each rearrangement produced a different array of segregation products at metaphase II, reflecting their contrasting frequencies of multivalent configurations at metaphase I. Comparison of chromosome behaviour at metaphase I and II suggested that certain configurations tended to adopt particular orientations. However, it was also clear that such correlations were imprecise and that other factors, possibly the exact positions of chiasmata, also played a role in multivalent orientation. Two rearrangements, T(2;4)1Go and T(7;16)67H, were analysed in female mice. The frequencies of the various multivalent types at metaphase I differed from those in male carriers of these rearrangements owing to an increased chiasma frequency in oocytes in some of the pairing segments. Not surprisingly, the segregation products seen in metaphase II cells showed some differences from the pattern recorded in male germ cells. For T(2;4)1Go, the sex-related difference in segregation patterns resulted in a diminished expectation of genetically imbalanced gametes, although this was not the case for T(7;16)67H. Received: 6 June 1998 / Accepted: 9 October 1998     

Meiotic events at the centromeric heterochromatin: histone H3 phosphorylation, topoisomerase IIα localization and chromosome condensation

Mechanisms of chromosome condensation and segregation during the first meiotic division are not well understood. Resolution of recombination events to form chiasmata is important, for it is chiasmata that hold homologous chromosomes together for their oppositional orientation on the meiotic metaphase spindle, thus ensuring their accurate segregation during anaphase I. Events at the centromere are also important in bringing about proper attachment to the spindle apparatus. This study was designed to correlate the presence and activity of two proteins at the centromeric heterochromatin, topoisomerase II alpha (TOP2A) and histone H3, with the processes of chromosome condensation and individualization of chiasmate bivalents in murine spermatocytes. We tested the hypothesis that phosphorylation of histone H3 is a key event instigating localization of TOP2A to the centromeric heterochromatin and condensation of chromosomes as spermatocytes exit prophase and progress to metaphase. Activity of topoisomerase II is required for condensation of chromatin at the end of meiotic prophase. Histone H3 becomes phosphorylated at the end of prophase, beginning with its phosphorylation at the centromeric heterochromatin in the diplotene stage. However, it cannot be involved in localization of TOP2A, since TOP2A is localized to the centromeric heterochromatin throughout most of meiotic prophase. This observation suggests a meiotic function for TOP2A in addition to its role in chromatin condensation. The use of kinase inhibitors demonstrates that phosphorylation of histone H3 can be uncoupled from meiotic chromosome condensation; therefore other proteins, such as those constituting metaphase-promoting factor, must be involved. These results define the timing of important meiotic events at the centromeric heterochromatin and provide insight into mechanisms of chromosome condensation for meiotic metaphase.     

Meiotic studies of male common shrews (Sorex araneus L.) from a hybrid zone between chromosome races

Thirty-three adult male common shrews (Sorex araneus L.) were collected from a hybrid zone between two chromosomal races that differed in Robertsonian metacentrics. Anaphase I nondisjunction frequencies were estimated on the basis of metaphase II counts. RIV and CV complex heterozygotes (four-element rings and five-element chains at meiosis I, respectively) had substantially higher nondisjunction rates than homozygotes and simple Robertsonian heterozygotes. However, at least in the case of RIV-forming hybrids, increased nondisjunction frequency did not result from malsegregation of the heterozygous complex. Extra elements found in hyperploid spreads were most frequently acrocentrics, that could not originate from a fully metacentric multivalent. Complex heterozygotes were also characterized by higher frequencies of univalents observed at diakinesis I. However, univalents did not originate from complex configurations, which were regularly formed with usually one chiasma per chromosome arm. Hence, we suppose that the presence of multivalents in the cell affects pairing and segregation of other elements at meiosis I.     

On the evolutionary stability of Mendelian segregation

We present a model of a primary locus subject to viability selection and an unlinked locus that causes sex-specific modification of the segregation ratio at the primary locus. If there is a balanced polymorphism at the primary locus, a population undergoing Mendelian segregation can be invaded by modifier alleles that cause sex-specific biases in the segregation ratio. Even though this effect is particularly strong if reciprocal heterozygotes at the primary locus have distinct viabilities, as might occur with genomic imprinting, it also applies if reciprocal heterozygotes have equal viabilities. The expected outcome of the evolution of sex-specific segregation distorters is all-and-none segregation schemes in which one allele at the primary locus undergoes complete drive in spermatogenesis and the other allele undergoes complete drive in oogenesis. All-and-none segregation results in a population in which all individuals are maximally fit heterozygotes. Unlinked modifiers that alter the segregation ratio are unable to invade such a population. These results raise questions about the reasons for the ubiquity of Mendelian segregation.     

Chromosomal identification and meiotic behavior of reciprocal translocations in a rye polymorphic population. Evolutionary implications

Summary The spontaneous interchange polymorphism of rye cultivar Ailés is composed, as can be deduced from the chromosomal identification of the interchanges analyzed, of several different reciprocal translocations in which the chromosomes of its haploid complement are involved with a similar frequency, except for chromosomes 4R and 6R. Several features of chromosome behavior at metaphase I, such as configuration and orientation of quadrivalents and frequency of chiasmata, were analyzed in structural heterozygotes for different interchanges. The two main factors affecting the orientation of quadrivalents at metaphase I proved to be the morphology of these chromosome associations at metaphase I and, in particular, the frequency of bound chromosome arms that they showed. A genotypic control of alternate orientation of quadrivalents independent of chiasmata frequency was not detected. In addition, the frequency of alternate orientation shows no relation to the fitness. Possible evolutionary implications of the results obtained are discussed.     

Spermatogenic delay and increased chiasma frequency in T70H/+ male mice with hydroxyurea-Trenimon limited spermatocyte populations

T(1;13)70H/+ male mice were treated with hydroxyurea (HU) and Trenimon (T). This karyotype offers excellent possibilities for estimating number and position of chiasmata and segregation in meiotic anaphase I. By their cell-killing action during spermatogenesis, HU and T produce large gaps in the spermatogenic line. The surviving population between the gaps was analysed at diakinesis--metaphase I and metaphase II. We found by autoradiography a considerable retardation of the development from resting primary spermatocytes (RPS) to metaphase I and II as compared to untreated T70H/+ males. Furthermore we found increased chiasma frequencies in diakinesis--metaphase I (MI) and reduced nondisjunction frequencies at anaphse I as a result of the treatments applied. The latter effect could not be explained by the increased chiasma frequency.