Chiasma localization,heterochromatin and synaptonemal complexes in the grasshopperPyrgomorpha conica

Surface-spread synaptonemal complexes and chiasma distributions in spermatocytes with different C-banding patterns and chiasma distributions in oocytes were analyzed in the grasshopperPyrgomorpha conica. Male meiosis was characterized by a proximal/distal chiasma localization and complete pairing of homologous chromosomes at pachytene. However, there were indications of a relationship between the frequency and location of pairing initiation sites and chiasma distribution. The presence of a proximal supernumerary segment in a medium-sized chromosome does not increase the mean cell chiasma frequency of carrier individuals compared with those lacking it but may modify chiasma distribution in at least some carrier bivalents. This effect could be related to heterosynapsis in the region near the segment. Mean cell chiasma frequency was significantly lower in females than in males. Females also showed altered chiasma distributions compared with males, with fewer proximal chiasmata and more interstitial and distal chiasmata.accepted for publication by M. Schmid     

Meiotic synapsis of homogeneously staining regions (HSRs) in chromosome 1 ofMus musculus

About 50 copies of a long-range repeat DNA family with a repeat size of roughly 100 kb and with sequence homology to mRNAs are clustered in the G-light band D of chromosome 1 of the house mouse,Mus musculus. We studied amplified versions of the cluster which are found in many wild populations ofM. musculus. They are cytogenetically conspicuous as one or two C-band positive homogeneously staining regions (single- and double band HSRs) which increase the mitotic length of chromosome 1. The double band HSR was phylogenetically derived from a single band HSR by a paracentric inversion. In homozygous condition, such HSRs contribute, albeit not as much as expected from their mitotic length, to the synaptonemal complex (SC) length of chromosome 1. In HSR heterozygous animals an elongation of the SCs was not noticeable. In single band HSR heterozygous males, synapsis proceeds regularly and continuously from the distal telomere towards the centromeric end without forming buckles. Thus, the single band HSR has no adverse effect on pairing. The same straight pairing behaviour was found in the majority of double band HSR heterozygous spermatocytes. This shows that extensive nonhomologous pairing can take place in the earliest phase of synapsis. Synapsis was discontinuous, leaving the central part of the bivalent 1 asynapsed, in only 14.3% of double band HSR heterozygous cells. In such cells the chromosome 1 SC is completed at a later stage of meiosis. The delay is presumably an effect of the inversion that includes one HSR band and the segment between the two HSR bands.     

Crossing over as assessed by late recombination nodules is related to the pattern of synapsis and the distribution of early recombination nodules in maize

Recombination nodules (RNs) are multicomponent proteinaceous ellipsoids found in association with the synaptonemal complex (SC) during prophase I of meiosis. Numerous early RNs (ENs) are observed during zygotene, and they may be involved in homologous synapsis and early events in recombination. Fewer late RNs (LNs) are observed during pachytene, and they occur at crossover sites. Here we describe the pattern of synapsis and the distribution of ENs and LNs in maize. Synapsis starts almost exclusively at chromosome ends, although later in zygotene there are many interstitial sites of synaptic initiation. ENs do not show interference, except possibly at distances ≤0.2 μm. The frequency of ENs is higher on distal compared to medial SC segments, and the highest concentration of ENs occurs at synaptic forks. The number of ENs on an SC segment does not change during zygotene. These observations are interpreted to indicate that ENs are assembled at synaptic forks. Like ENs, LNs are more concentrated distally on bivalents but, unlike ENs, LNs show interference. A model is presented that relates the pattern of synapsis and ENs to the pattern of late nodules and crossing over. This revised version was published online in July 2006 with corrections to the Cover Date.     

Asynchronous chromosome pairing in male meiosis of the rat (Rattus norvegicus)

Premeiotic and meiotic chromosome distribution was studied in rat testes suspensions by a triple-color fluorescent staining protocol which allows simultaneous visual inspection of two chromosomal targets highlighted by FISH together with immunostained SCP3 synaptonemal complex (SC) proteins which are marked by a third, composite color. Triple labeling with rat chromosome (RNO) 4q and 19p specific probes and SCP3 staining disclosed that homologs are separated in premeiotic and leptotene nuclei. Pairing of homologous chromosome regions commenced during early zygotene, with pairing of the small metacentric chromosomes 19 preceding that of the distal region of the long arm of RNO4. Our results show that homolog association occurs during zygotene of rat spermatogenesis, with small and large chromosomes showing a considerable asynchrony. Comparison with pairing progression in meiosis of other mammals suggests that asynchronous chromosome pairing reflects size differences within a complement. This revised version was published online in July 2006 with corrections to the Cover Date.     

Chiasma formation in Arabidopsis thaliana accession Wassileskija and in two meiotic mutants

Meiotic chiasmata were analysed in metaphase I pollen mother cells (PMCs) of wild-type Arabidopsis thaliana and in two meiotic mutants. Fluorescence in situ hybridisation (FISH) with 45S rDNA and 5S rDNA as probes was used to identify the five chromosome pairs. A wild-type chiasma frequency of 9.24 per cell was found, consistent with estimated genetic recombination values. Individual bivalent chiasma frequencies varied according to chromosome size; chromosome 1 had the highest mean chiasma frequency (2.14) while the short acrocentric chromosomes had the lowest frequencies (1.54 and 1.56). FISH analysis was extended to two meiotic mutants (asy1 and dsy1) having low residual bivalent and chiasma frequencies. Mutant dsy1 gave no indication of chromosome preference for residual bivalent formation; instead it showed a general reduction in bivalent and chiasma frequencies. In asy1, the longest chromosome (1) had the lowest bivalent frequency and chiasma frequency while the short acrocentric chromosome 2 had the highest frequencies. This chromosome pair may be preferentially involved in synapsis and chiasma formation because of their association with the nucleolus. However, other factors may be operating since the other acrocentric chromosome (4), with similar size and structure to chromosome 2, did not share these chiasma properties.     

Meiotic behaviour of holocentric chromosomes: orientation and segregation of autosomes in Triatoma infestans (Heteroptera)

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 hetero-chromatic 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 orheterochromatic). 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. This revised version was published online in August 2006 with corrections to the Cover Date.     

Telomere distribution pattern and synapsis initiation during spermatogenesis in zebrafish

Background: Telomeres are located at ends of eukaryotic chromosomes and can affect proper chromosomal positioning. During spermatogenesis, the appropriate dynamics and behavior of chromosomes is crucial to generate haploid cells through meiosis. Here, we describe telomere distribution patterns during spermatogenesis in zebrafish, especially during meiotic prophase I, using fluorescence in situ hybridization. This was combined with synaptonemal complex protein 3 immunostaining, which allows the staging of spermatocytes. Results: During spermatogonial proliferation and the preleptotene stage, telomeres were dispersed throughout the nucleus. During the leptotene stage, telomeres temporarily moved to one pole of the nucleus at which γ‐tubulin was located, forming the telomere bouquet. The cluster lasted until the onset of zygotene where it coincided with terminal synapsis initiation. They then spread around the periphery of the nucleus during the zygotene to pachytene stages. During postmeiotic stages, telomeres in spermatids and sperm were again dispersed throughout the nuclei. Application of this procedure in meiotic mutants confirmed that meiotic telomere clustering is independent of axial element formation of the synaptonemal complex. Conclusions: These data clearly showed the clustering and distributions of telomeres throughout spermatogenesis in zebrafish. This procedure could be used to screen for mutants that have primary defects in telomere clustering. Developmental Dynamics 243:1448–1456, 2014. © 2014 Wiley Periodicals, Inc.     

Synaptonemal complex analysis in spermatocytes and oocytes of rainbow trout,Oncorhynchus mykiss (Pisces,Salmonidae): the process of autosome and sex chromosome synapsis

The surface-spreading synaptonemal complex (SC) technique was employed to analyze spermatocytes and oocytes of rainbow trout in order to visualize the process of autosome and sex chromosome synapsis in this species. The structure of lateral elements (LEs) of the SC and the chromosome synapsis process at the stages of leptotene, zygotene and pachytene are described. Comparative analysis of SCs of spermatocytes and oocytes showed a difference in the synaptic process, i.e. in spermatocytes all LEs were synapsed before the appearance of centromeric regions in the biarmed elements, while in the oocytes some fully synapsed LEs, including the centromeric region of the biarmed elements, were found together with fully or partially unsynapsed LEs. In males the sex chromosome synapsis starts only after all autosomes have synapsed. Irregular synapses involving three or four LEs were found in 3.4% of the cells analyzed in mid or late zygotene. Multivalents were found in males and females. Some aspects of initial meiotic development and their implications in rainbow trout cytogenetics, genetics and evolution are discussed.     

Synaptonemal polycomplexes in spermatids: a characteristic trait of Orthoptera?

Spermatogenesis was analysed in a cricket,Eneoptera surinamensis (Gryllidae, Orthoptera), using ultrathin serial sections and transmission electron microscopy. Special attention was placed on documentation of the development and structure of synaptonemal polycomplexes (PCs) within spermatid nuclei. Pachytene spermatocytes showed the usual tripartite synaptonemal complexes in the nuclear lumen. PCs were situated close to chromosomes at the periphery of spindles in prometaphase I spermatocytes, where microtubule density was low. The PCs are probably incorporated into the daughter nuclei of both meiotic divisions by adhesion to chromosomes. Finally, PCs end up within spermatid nuclei. Analysis of serial sections through three nuclei of young spermatids revealed at least one PC within each. The PCs were intimately attached to an electrondense spherical nuclear body. This topographical correlation was confirmed through inspection of random sections. The PCs may have an affinity to the spherical bodies. In more developed spermatids, PCs and nuclear bodies were missing. Disassembly products of the PCs may play a role in spermatid maturation. In a series of other Orthoptera species, PCs have been reported to occur in the cytoplasm or the nuclei of spermatids. In most other systematic groups, PCs do not form at all or disassemble earlier. The presence of PCs in young spermatids, therefore, seems to be typical of Orthoptera.     

Regional localization of two MRX genes to Xq28 (MRX28) and to Xp11.4-Xp22.12 (MRX33)

Two genes responsible for a nonspecific form of X-linked mental retardation (MRX28 and MRX33) were localized by linkage analysis with 40 highly polymorphic DNA markers situated along the entire the X chromosome. In family 1, the gene could be mapped within a 14-cM interval at Xq28, distal to the recombining marker DXS1113 (MRX28). The maximum LOD score was 2.75, with DXS52 at ϕ = .0. In family 2, the gene was localized within a 30-cM interval at Xp11.4-22.12 between the recombining markers DXS365 and MAOB, including the DMD gene (MRX33). Maximum LOD scores of 2.82 were obtained with markers DMD-STR49, DMD-DysII, CYBB, and DXS1068. © 1996 Wiley-Liss, Inc.     

Necrotizing (malignant) otitis externa: an unusual localization of mucormycosis

Malignant otitis externa (MOE) is a severe infection of external auditory canal and skull base. A 17-year-old diabetic girl was admitted with diabetic ketoacidosis. Cellulitis of her right ear occurred on the second day of hospitalization and a black necrotic scar in the same region appeared on the next day. The lesion rapidly invaded to right side of neck and surrounding tissue of the patient. Therefore, antimycotic therapy was started. Unfortunately the patient died on seventh day of hospitalization because of probably extensive fungal invasion. Physicians should suspect MOE connected to mucormycosis especially in patients with cutaneous lesions of ear unresponsive to antibiotic therapy.     

Microsurgical anatomy of the pituitary gland (hypophysis cerebri) and the sellar region. Part IV: The optic nerves and optic chiasma

The anatomical relationships of the optic nerve and optic chiasma to the different structures of the sellar region were studied in 100 cadaver sphenoidal blocks and in patients during transfrontal surgery to the sellar region. This study includes the relationships with the bony structures (tuberculum sellae, dorsum sellae, sella turcica, optic canal), with the meninges (arachnoidal cisterns, tentorium of the optic nerve), with the vessels (carotid and ophthalmic arteries), and finally with the neural structures (hypophysis cerebri, cranial nerves third ventricle). Relevant clinical or surgical aspects in relation to normal anatomy and anatomical variations of the optic nerves and optic chiasma are discussed. The varieties of the chiasma (normal, prefixed, postfixed) and the measurements of the optic nerves and optic chiasma (width, length, height, distance, and angle between optic nerves) were studied in the cadaver only. Different transfrontal approaches to the sellar region are discussed according to the morphology of the chiasma.     

Source localization using eventrelated desynchronization (ERD) within the alpha band

Summary Voluntary finger movements result in a maximal ERD in the 10–12 Hz band close to electrodes C₃ and C₄, overlying the sensorimotor hand areas. This ERD focus is not very pronounced with EEG data recorded against a common reference electrode (monopolar recording). After transformation of the raw data using common average, local average and weighted average reference and the Hjorth method, respectively, the ERD becomes enhanced over electrodes C₃ or C₄. Movement-related EEG data were studied with 17, 19, 30 and 56 electrode montages using large and small interelectrode distances. The best focused ERD was obtained with a 56 electrode montage with small interelectrode distances and local average reference data.Supported by the Fonds zur Förderung der wissenschaftlichen Forschung project S49/02 and the Österreichische Nationalbank, project 4534. We thank J. Kalcher for support in programming and W. Mohl for data aquisition.     

Pathogenic variants in the paired-related homeobox 1 gene (PRRX1) cause craniosynostosis with incomplete penetrance

PurposeStudies have previously implicated PRRX1 in craniofacial development, including demonstration of murine Prrx1 expression in the preosteogenic cells of the cranial sutures. We investigated the role of heterozygous missense and loss-of-function (LoF) variants in PRRX1 associated with craniosynostosis.MethodsTrio-based genome, exome, or targeted sequencing were used to screen PRRX1 in patients with craniosynostosis; immunofluorescence analyses were used to assess nuclear localization of wild-type and mutant proteins.ResultsGenome sequencing identified 2 of 9 sporadically affected individuals with syndromic/multisuture craniosynostosis, who were heterozygous for rare/undescribed variants in PRRX1. Exome or targeted sequencing of PRRX1 revealed a further 9 of 1449 patients with craniosynostosis harboring deletions or rare heterozygous variants within the homeodomain. By collaboration, 7 additional individuals (4 families) were identified with putatively pathogenic PRRX1 variants. Immunofluorescence analyses showed that missense variants within the PRRX1 homeodomain cause abnormal nuclear localization. Of patients with variants considered likely pathogenic, bicoronal or other multisuture synostosis was present in 11 of 17 cases (65%). Pathogenic variants were inherited from unaffected relatives in many instances, yielding a 12.5% penetrance estimate for craniosynostosis.ConclusionThis work supports a key role for PRRX1 in cranial suture development and shows that haploinsufficiency of PRRX1 is a relatively frequent cause of craniosynostosis.     

Ultrastructural localization of basic proteins and carbohydrates in male accessory glands of two Triatoma species (Hemiptera, Reduviidae, Triatominae)

The male accessory glands in Triatoma are tubular and produce substances with some functions related to production of the spermatophore. In the current study, the cytochemistry of male accessory glands was evaluated in starved Triatoma brasiliensis and adult Triatoma melanica. The storage of carbohydrates and proteins in T. melanica male accessory glands occurs earlier than in T. brasiliensis. In addition, the occurrence of eletron-lucent granules without carbohydrates and proteins suggests that other substances are released by these glands, which may be used for lubrication of the male genitalia. Male T. brasiliensis has more intense secretory activity in the fifth day of adult life, which may indicate a higher reproductive capacity. The analysis of lipid production in male accessory glands can contribute to the knowledge of spermatophore formation in these species.     

Behavioral coupling in tettigoniid hybrids (Orthoptera)

Studies of the mating behavior of male and female F₁ hybrids between closely related taxa can provide information concerning the genetic control of characters that play a major role in speciation. Orthoptera have been used previously for such studies. Hybrid crickets show behaviors which are broadly intermediate to the parentals but hybrid grasshoppers may retain parental behavior patterns. This study examines the behavior of hybridEphippiger ephippiger bushcrickets, the third major orthopteran group. The differences in male song and female preference are probably both mainly additive and male song differences not sex linked. Thus, given a choice, hybrid females would prefer to mate with hybrid males, an example of behavioral coupling. The evolutionary inferences which can be drawn from studies of F₁ hybrids between closely related taxa are discussed.This work took place during a U.K. Science and Engineering Research Council postdoctoral research fellowship.     

Cellular localization of iron(II) polypyridyl complexes determines their anticancer action mechanisms

Elucidation of relationship among cellular uptake, localization and biological activities of metal complexes could make great breakthrough in the understanding of their action mechanisms and provide useful information for rational design of metal-based anticancer drugs. Iron(II) complexes have emerged as potential anticancer drug candidates with application potential in cancer imaging and therapy. Herein, a series of iron(II) polypyridyl complexes with different lipophilicity were rationally designed, synthesized and identified as potent anticancer agents. The relationship between the cellular localization and molecular action mechanisms of the complexes was also elucidated. The results showed that, the increase in planarity of the Fe(II) polypyridyl complexes enhanced their lipophilicity and cellular uptake, leading to improved anticancer efficacy. The hydrophilic Fe(II) complex entered cancer cells through transferring receptor (TfR)-mediated endocytosis, and translocated to cell nucleus, where they induced S phase cell cycle arrest through triggering DNA damage-mediated p53 pathway. Interestingly, the hydrophobic Fe(II) complexes displayed higher anticancer efficacy than the hydrophilic ones, but shared the same uptake pathway (TfR-mediated endocytosis) in cancer cells. They accumulated and localized in cell cytoplasm, and induced G0/G1 cells cycle arrest through regulation of AKT pathway and activation of downstream effector proteins. These results support that the cellular localization of Fe(II) complexes regulated by their lipophilicity could affect the anticancer efficacy and action mechanisms. Taken together, this study may enhance our understanding on the rational design of the next-generation anticancer metal complexes.     

Synapsis and chiasma formation in four meiotic mutants of tomato (Lycopersicon esculentum)

Synapsis and chiasma formation were studied in pollen mother cells of four meiotic mutants of tomato. The four mutants displayed defects in the assembly of the synaptonemal complex (SC) covering the whole range from almost complete absence of synapsis to complete synapsis at pachytene. In three mutants, we found a good correlation between the number of bivalents connected by at least one tripartite SC segment at pachytene and the number of chiasmatic bivalents at metaphase I. We suggest that in tomato functional chiasmata are only formed in the context of the tripartite SC.