Large-scale physical mapping within the region 22q12.3-13.1 in meningioma

The lack of physical mapping data strongly restricts the analysis of the meningioma chromosomal region that was assigned to the bands 22q12.3-qter. Recently, we reported a new marker D22S16 for chromosome 22 that was assigned to the region 22q13-qter by in situ hybridization. Utilizing somatic cell hybrids we now sublocalized the marker D22S16 within the band region 22q12-13.1, thus placing it in the vicinity of the gene for the platelet derived growth factor (PDGFB). A physical map was established for the regions surrounding the PDGFB gene and the D22S16 marker. By means of pulsed-field gel electrophoresis (PFGE) D22S16 and PDGFB were found to be physically linked within 900 kb. We also identified two CpG clusters bordering the PDGFB gene. For the enzyme NotI, a variation of the PDGFB restriction pattern was found between different individuals. PFGE analysis of the two loci (PDFGB and D22S16) failed to identify major rearrangements in meningioma.     

Chromosome organization revealed upon the decondensation of telophase chromosomes inAllium

Chromosomes of root tip cells ofAllium cepa andAllium sativum were studied in early, middle and late telophase to examine the organization of mitotic chromosomes, taking advantage of the naturally occurring chromosome dispersion during the process of decondensation in telophase. Longitudinal and transverse sections of telophase chromosomes viewed under the transmission electron microscope showed that mitotic chromosomes inAllium were composed of helically coiled 400–550 nm chromatin fibres. In some regions of the longitudinal sections, these chromatin fibres were seen to be orientated parallel to one another but formed roughly a right angle to the long axis of the chromosome. In transverse sections, the telophase chromosome appeared to have a hollow centre encircled by the 400–550 nm chromatin fibre which in turn was a hollow tube structure formed by the coiling of a thinner fibre of 170–200 nm. In addition, cross views of chromatin fibres of 170–200 nm and 50–70 nm were also identified in telophase chromosome preparations. These two organizational levels of chromatin fibres also showed a hollow centre. The process of decondensation of telophase chromosomes is described, and some morphological characteristics associated with the activities of chromosome decondensation are analysed. Based on the observations made onAllium chromosomes in this study, various models of chromosome organization are discussed.     

Quantitative studies on the arrangement of human metaphase chromosomes

Summary The spatial relationships between the homologous pairs of chromosomes in the normal human colcemid-treated metaphase plate were tested by two different mathematical approaches: (a) determination of the distances between the centromeres of the homologous chromosomes compared to the mean distance of all centromeres of the mitosis in question; (b) measuring the distances of the different chromosomes from the center of the mitosis.The following results were obtained: (1) The arrangement of human metaphase chromosomes does not follow a normal distribution; the distribution is narrower and taller, probably due to an impairment of free chromosome spreading by the cell membrane. We believe that only in membraneless mitotic cells should the chromosome-spread correspond to a normal distribution under the same preparation conditions. (2) There is a positive correlation between decreasing chromosome size and decreasing mean distance between homologous chromosomes. (3) A close positive correlation exists between increasing chromosome size and increasing distance to the barycenter of the mitosis. (4) There is also a close positive correlation between the distance of homologous chromosome pairs and their distance from the center of the mitosis, i.e., with increasing distance from the center of the mitosis, the distance between the homologous partners increases. (5) The following statistically significant deviations from these rules could be established: (a) The large acrocentric chromosomes are closer associated, as one would expect from their size, probably due to their participation in the nucleolus organization; (b) in the female cell one of the two X chromosomes has an extremely peripheral localization; the X chromosomes are furthest apart of all pairs of homologous chromosomes; (c) the chromosome pairs 6 and 8 are relatively close together in spite of their peripheral position, suggesting a truc close association of the homologus partners; (d) the chromosome pair 18 has a more peripheral position than expected, and a relatively large mean distance between the homologous partners; (e) the chromosome pair 1 has a much more central position and a closer association than is expected from its size.     

Quantitative studies on the arrangment of human metaphase chromosomes

Summary The pattern of association of acrocentric chromosomes was examined in ten and five carriers of a 15/21 and a 13/14 Robertsonian translocation, respectively, and was compared with that of the same numbers of relatives with normal karyotypes. In the carriers of 15/21 translocation, the number of large associations (involving more than two acrocentrics) and the association frequencies for individual acrocentric chromosomes, were significantly higher than in the control group. The mean number of associations of the single homologs of the translocation chromosomes was much higher than that of the other acrocentrics. In the carriers of 13/14 translocations, only the association frequency for chromosome 13 was higher than in the normal relatives. The uninvolved chromosomes homologous to those involved in translocations showed an insignificant increase in associations in comparison with the other acrocentrics. These results suggest that some mechanism within the cells compensates for the effect of missing acrocentrics or of acrocentrics lacking NORs on the number of associations. The possible relations of this phenomenon to the activity of the nucleolus organizing regions are discussed.     

Cytological and cytogenetical studies on brain tumors. V. Preferential loss of sex chromosomes in human meningiomas.

Twelve out of 88 cytogenetically examined meningiomas of female patients showed, in addition to the typical loss of a chromosome 22, a loss of 1 or more chromosomes of group C. Among them 8 tumors had less than 8% cells with Barr-body-like particles, whereas in one tumor 12% and in 3 others over 20% Barr bodies were found, which, based on control studies, were classified as sex-chromatin negative, partly positive, and positive, respectively. In one case the loss of an X chromosome was verified by Giemsa banding. In 6 out of 24 meningiomas of male origin, the chromosomal morphology and association pattern strongly indicated that besides the loss of a chromosome 22, the Y chromosome was also missing. Moreover, the loss of the male sex chromosome could be ascertained in 4 tumors by the conspicuous absence of Y fluorescence in interphase nuclei and in metaphase plates after fluorescence staining. The findings are discussed in connection with the gonosomal loss in other human tumors and in old age.     

Cytological and cytogenetical studies on brain tumors

Summary Twelve out of 88 cytogenetically examined meningiomas of female patients showed, in addition to the typical loss of a chromosome 22, a loss of 1 or more chromosomes of group C. Among them 8 tumors had less than 8% cells with Barr-body-like particles, whereas in one tumor 12% and in 3 others over 20% Barr bodies were found, which, based on control studies, were classified as sex-chromatin negative, partly positive, and positive, respectively. In one case the loss of an X chromosome was verified by Giemsa banding.In 6 out of 24 meningiomas of male origin, the chromosoma. morphology and association pattern strongly indicated that besides the loss of a chromosome 22, the Y chromosome was also missing. Moreover, the loss of the male sex chromosome could be ascertained in 4 tumors by the conspicuous absence of Y fluorescence in interphase nuclei and in metaphase plates after fluorescence staining.The findings are discussed in connection with the gonosomal loss in other human tumors and in old age.

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Zusammenfassung Unter 88 cytogenetisch untersuchten Meningeomen von Frauen wurden 12 Tumoren gefunden, bei denen außer dem für Meningeome typischen Verlust eines Chromosoms 22 auch ein oder mehrere Chromosomen der C-Gruppe verlorengegangen waren. Bei 8 dieser Tumoren konnte in Gewebekulturpräparaten nur in weniger als 8% der untersuchten Zellen Barr-body-ähnliche Kernstrukturen nachgewiesen werden, bei einem Tumor fanden sich 12% und bei 3 über 20% Barr-bodies. Auf Grund von Vergleichsuntersuchungen wurden 8 Tumoren als geschlechtschromatinnegativ, 1 Tumor als teilweise positiv und die übrigen 3 als eindeutig positiv eingestuft. Bei einem Meningeom konnte das Fehlen eines X-Chromosoms direkt mit der Giemsa-Bandentechnik nachgewiesen werden.Bei 6 von 24 Meningeomen männlicher Herkunft konnte auf Grund der Chromosomenmorphologie und des Assoziationsverhaltens sehr wahrscheinlich gemacht werden, daß außer dem Chromosom 22 auch das Y-Chromosom verlorengegangen war. Bei 4 dieser Tumoren konnte eine Fluorescenzfärbung durchgeführt werden, wobei das Fehlen einer Y-Fluorescenz in Interphasezellen und Metaphaseplatten nachweisbar war.Diese Befunde werden im Zusammenhang mit dem Geschlechtschromosomenverlust bei anderen menschlichen Tumoren und im hohen Lebensalter diskutiert.

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Supported by the Deutsche Forschungsgemeinschaft (SFB 51 E 12).

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Parts of this work are included in the doctoral thesis (M.D.) of H.S. at the University of Munich, Germany.     

A mechanism coordinating root elongation,endodermal differentiation,redox homeostasis and stress response

Root growth relies on both cell division and cell elongation, which occur in the meristem and elongation zones, respectively. SCARECROW (SCR) and SHORT-ROOT (SHR) are GRAS family genes essential for root growth and radial patterning in the Arabidopsis root. Previous studies showed that SCR and SHR promote root growth by suppressing cytokinin response in the meristem, but there is evidence that SCR expressed beyond the meristem is also required for root growth. Here we report a previously unknown role for SCR in promoting cell elongation. Consistent with this, we found that the scr mutant accumulated a higher level of reactive oxygen species (ROS) in the elongation zone, which is probably due to decreased expression of peroxidase gene 3, which consumes hydrogen peroxide in a reaction leading to Casparian strip formation. When the oxidative stress response was blocked in the scr mutant by mutation in ABSCISIC ACID 2 (ABA2) or when the redox status was ameliorated by the upbeat 1 (upb1) mutant, the root became significantly longer, with longer cells and a larger and more mitotically active meristem. Remarkably, however, the stem cell and radial patterning defects in the double mutants still persisted. Since ROS and peroxidases are essential for endodermal differentiation, these results suggest that SCR plays a role in coordinating cell elongation, endodermal differentiation, redox homeostasis and oxidative stress response in the root. We also provide evidence that this role of SCR is independent of SHR, even though they function similarly in other aspects of root growth and development.