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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Chromosoma 107 (1998), S. 127-135 
    ISSN: 1432-0886
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Small chromosomal deletions [Df(3R)e R−1 and Df(3R)e P ] with intact hsrω transcription units but with variable deletions of the upstream region were used to map the upstream regions that regulate heat shock and amide responsivity of the 93D puff (hsrω locus) in salivary glands of late third instar larvae of Drosophila melanogaster. The Df(3R)e P deletion, generated by a P-element mobilization screen, removed the 93B6–7 to 93D3–5 cytogenetic region. [3H]uridine-labeled transcription autoradiograms revealed that normal developmental and heat shock-induced expression of the 93D puff remained unaffected in both the deficiency chromosomes. However, the amide responsivity of the 93D site was lost on the Df(3R)e P homolog while the Df(3R)e R−1 homolog responded normally to amides. Southern hybridizations with a series of upstream probes mapped the distal breakpoint of the Df(3R)e P deletion between −22 kb and −23 kb of the hsrω transcription unit. Since the distal breakpoint of Df(3R)e R−1 is at about −45 kb upstream of the hsrω gene it is inferred that the amide response element(s) that modulate the specific transcriptional activation of the 93D puff following treatment of salivary glands with a variety of amides is/are located in the −22 kb to about −45 kb upstream interval. The Df(3R)e P and Df(3R)e R−1 deletions also abolished dosage compensation at the 93D locus as well as the effect of β-alanine levels on its heat shock inducibility.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Chromosoma 89 (1984), S. 63-67 
    ISSN: 1432-0886
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract It is widely known that the bulk of the pericentromeric heterochromatin (α-heterochromatin) does not replicate during polytenization in Drosophila. However, a recent DNA-Feulgen cytophotometric study (Dennhöfer 1982a) has claimed equal polytenization of all heterochromatin regions. To re-examine this issue, the amount of Hoechst 33258-bright heterochromatin in non-polytene and polytene nuclei in salivary glands and Malpighian tubules of late third instar larvae of D. nasuta has been compared by cytofluorometry. Since the amount of Hoechst 33258-bright heterochromatin is similar in non-polytene and polytene nuclei in spite of the latter having an enormously high euchromatin DNA content, it is concluded that the α-heterochromatin does not replicate during polytenization. The present results further indicate that in the polytene nuclei of Malpighian tubules the α-heterochromatin remains at the 2C level whereas in salivary gland polytene nuclei it varies between the 2C and 4C levels.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Chromosoma 89 (1984), S. 212-217 
    ISSN: 1432-0886
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The temporal order of replication of specific sites in polytene chromosomes from salivary glands and gastric caeca of Drosophila nasuta larvae was compared using 3H-thymidine autoradiography. Labelling of different cytological regions in segments of chromosome 2R (section 47 A to 49 C) and chromosome 3 (section 80 A to 82 C) was examined in detail in nuclei showing late S-period labelling (2 D and 1D types) in both cell types. The different labelling sites (22 on the 2R segment and 38 on the chromosome 3 segment) are cytologically similar in the two cell types. However, there are profound differences in the labelling frequencies of certain sites in polytene nuclei from salivary glands and gastric caeca during the late S-phase. This suggests that even though a comparable number of chromosomal replicating units operates in the two polytene cell types, the temporal order of completion of replication differs.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Genetica 97 (1996), S. 339-348 
    ISSN: 1573-6857
    Keywords: benzamide ; colchicine ; cytoskeleton ; heat shock ; puffs
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The 93D, or hsr-omega (heat-shock RNA-omega), locus of Drosophila melanogaster and other species of Drosophila, besides being induced as a member of the heat shock gene family, is also selectively and singularly inducible by a variety of agents, notably benzamide, colchicine and vitamin B6 (in species other than D. melanogaster). The genomic structure of this locus is highly conserved in all species, although the primary base sequence has diverged rapidly between species. Three transcripts (two nuclear and one cytoplasmic) are produced by this locus but none of them has any significant protein coding capacity. The profile of the three transcripts varies in a developmental and inducer-specific manner. This locus is developmentally active in nearly all cell types and is essential for viability of flies. Its induction during heat shock is independent of the other members of the heat shock gene family. The other selective inducers act on this locus through separate response elements. hsr-omega activity has a characterstic effect on transcription/turnover of the heat shock induced hsp70 and the alpha-beta transcripts in D. melanogaster. It appears that the hsr-omega locus has important house-keeping functions in transport and turnover of some transcripts and in monitoring the ‘health’ of the translational machinery of the cell.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-0886
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract A simple fluorescence technique is decribed to localise in situ the non-replicating alpha heterochromatin in the chromocentre region of Drosophila nasuta polytene nuclei. After incorporating 5-bromodeoxyuridine in larval salivary gland cells for one or two cycles of replication, the polytene nuclei are examined for Hoechst 33258 flourescence at pH 7.O. The nonreplicating alpha heterochromatin remains brightly fluorescing as it does not incorporate any 5-bromodeoxyuridine while the rest of the replicating chromatin shows dull fluorescence due to the quenching of Hoechst 33258 fluorescence by the bromodeoxyuridine substituted DNA.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1432-0886
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Larval brain ganglia of Drosophila nasuta were cultured in vitro in the presence of 5-bromodeoxyuridine for 1 or 5 h at 24° C and the air-dried chromosome preparations stained by the Hoechst 33258-Giemsa technique to reveal bromodeoxyuridine induced sister chromatid differentiation. In 1 h as well as 5 h preparations, 10–15% of well spread metaphase plates show a sister chromatid differentiation in only C-band heterochromatin regions of different chromosomes. We infer that this sister chromatid differentiation in all heterochromatic regions is seen after bromodeoxyuridine incorporation for only one replication cycle and is related to the presence of asymmetric A-T rich satellite sequences in all the C-band regions of D. nasuta karyotype.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1432-0886
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The autoradiographic patterns of 3H-uridine labelling of the major temperature shock puff sites and the chromocentric (β-)heterochromatin in heat shocked (37° C) salivary glands of Drosophila melanogaster have been studied. It is seen that in response to the heat shock treatment, four of the major temperature shock puffs (63BC, 87A, 87C and 95D) show a correlated level of 3H-uridine incorporation in a given nucleus. However, although the mean grain density on 93D puff is maximum in the heat shocked preparations, in individual nuclei this puff is labelled to a widely varying level and this variation in its labelling is statistically not correlated to the labelling of the other four temperature shock puffs in a nucleus. The chromocentric β-heterochromatin, which has been shown in several earlier studies to hybridize with temperature shock RNA fractions, is seen to be totally inactive in transcription after the heat shock.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1432-0886
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract A 10 or 20 min in vitro treatment of salivary glands of late 3rd instar larvae of Drosophila melanogaster with l mg/ml benzamide (BM) at 24° C results in the specific induction of the 93 D puff and at the same time all other chromosomal RNA synthesis is severely repressed. Incorporation of 3H-uridine in the nucleolus is not affected. In glands heat shocked (37° C) for 20 min in presence of BM, all the temperature shock (TS) puffs are induced but they incorporate 3H-uridine to a lesser extent than in glands heat shocked in absence of BM. The 93 D puff, which is highly induced by either of the treatments alone, is relatively less active in glands exposed to TS and BM simultaneously. When a 10 min BM treatment (at 24° C) precedes a 20 min TS or when the BM treatment follows TS, the 3H-uridine incorporation on all TS puffs is relatively less and significantly, in both cases, the 87 C puff is much less active (more so in TS followed by BM treated glands) than the 87 A puff. Also, in both these treatments, the 93 D puff does not show any additive effect of the BM and TS treatments. These observations are discussed in the light of possible role of the 93 D puff in modulating the heat shock response.
    Type of Medium: Electronic Resource
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Chromosoma 85 (1982), S. 221-236 
    ISSN: 1432-0886
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Prolongation of larval life in Drosophila melanogaster, by growing wild type larvae at lower temperature, or in animals carrying the X-linked mutation giant is known to result in a greater proportion of nuclei in salivary glands showing the highest level of polyteny. We have examined by autoradiography the patterns of 3H-thymidine incorporation during 10 min or 1 min pulses in salivary gland polytene chromosomes of older giant larvae and of wild type late third instar larvae of D. melanogaster grown since hatching either at 24 ° C or at 10 ° C. The various patterns of labelling and their relative frequencies are generally similar in glands from the warm-(24 ° C) or cold (10 ° C)-reared wild type larvae, except the interband (IB) labelling patterns which are very frequent in the later group but rare in the former. The IB type labelled nuclei in cold-reared wild type larvae show labelling ranging from only a few puffs/interbands labelled to nearly all puffs/interbands labelled. In warm-reared wild type larvae, very low labelled IB patterns are not seen. In older giant larvae, the 3H-thymidine labelling patterns are in most respects similar to those seen in cold-reared wild type larvae. In 1 min pulsed preparations from all larvae, the IB patterns are relatively more frequent than in corresponding 10 min pulsed preparations. No nuclei with the continuous (2C or 3C) type of labelling pattern, with all bands and interbands/puffs labelled, were seen in 1 min pulsed preparations from cold-reared wild type or in giant larvae, and only a few nuclei in 1 min pulsed preparations from warm-reared wild type larvae exhibited the 2C labelling pattern. Analysis of silver grain density on specific late replicating sites in late discontinuous (1D) type labelled nuclei suggests that the rate of DNA synthesis per chromosomal site is not different at the two developmental temperatures. It is suggested that correlated with the prolongation of larval life under cold-rearing conditions or in giant larvae, the polytene replication cycles are also prolonged. It is further suggested that the polytene S-period in these larvae is longer due to a considerable asynchrony in the initiation and termination of replication of different sites during a replication cycle.
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 1432-0886
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Salivary glands of Drosophila larvae were treated in vitro with benzamide or with a homogenate of heat shocked glands to specifically induce high transcriptional activity of the 93D puff. The newly synthesized 14C-amino acids labelled polypeptides in the treated and sister control glands were analysed by polyacrylamide gel electrophoresis, followed by gel autoradiography. The protein synthesis patterns in the treated glands in either case remain the same as in control glands. No novel polypeptide was seen which could be correlated with the high induced transcriptional activity of the 93D puff. This suggests that the 93D transcript/s is/are probably not translated.
    Type of Medium: Electronic Resource
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