Effect of 5-bromodeoxyuridine on pupariation and adult differentiation in the flesh fly, Sarcophaga bullata

1984 ◽  
Vol 62 (2) ◽  
pp. 157-160
Author(s):  
P. Sivasubramanian
Keyword(s):  
Imaginal Discs ◽  
Flesh Fly ◽  
Biochemical Processes ◽  
Sarcophaga Bullata ◽  
In Vivo Culture ◽  
Two Stages ◽  
Third Instar Larvae

Two stages of postfeeding, third-instar larvae of Sarcophaga bullata were injected with 5-bromodeoxyuridine to examine the effect of the analogue on pupariation and adult differentiation. While pupariation was inhibited in prered spiracle-stage larvae, adult differentiation was unaffected in both stages as revealed by in vivo culture of imaginal discs. It is suggested BudR may be more effective in disturbing certain biochemical processes than in inhibiting morphogenesis.

Hedgehog is required for activation of engrailed during regeneration of fragmented Drosophila imaginal discs

Development ◽  
1999 ◽  
Vol 126 (8) ◽  
pp. 1591-1599 ◽  
Author(s):  
M.C. Gibson ◽  
G. Schubiger
Keyword(s):  
Normal Growth ◽  
Imaginal Discs ◽  
Ideal Model ◽  
Posterior Compartment ◽  
Anterior Compartment ◽  
In Vivo Culture ◽  
Pattern Regulation ◽  
Regulative Capacity ◽  
Anterior Posterior

Surgically fragmented Drosophila appendage primordia (imaginal discs) engage in wound healing and pattern regulation during short periods of in vivo culture. Prothoracic leg disc fragments possess exceptional regulative capacity, highlighted by the ability of anterior cells to convert to posterior identity and establish a novel posterior compartment. This anterior/posterior conversion violates developmental lineage restrictions essential for normal growth and patterning of the disc, and thus provides an ideal model for understanding how cells change fate during epimorphic pattern regulation. Here we present evidence that the secreted signal encoded by hedgehog directs anterior/posterior conversion by activating the posterior-specific transcription factor engrailed in regulating anterior cells. In the absence of hedgehog activity, prothoracic leg disc fragments fail to undergo anterior/posterior conversion, but can still regenerate missing anterior pattern elements. We suggest that hedgehog-independent regeneration within the anterior compartment (termed integration) is mediated by the positional cues encoded by wingless and decapentaplegic. Taken together, our results provide a novel mechanistic interpretation of imaginal disc pattern regulation and permit speculation that similar mechanisms could govern appendage regeneration in other organisms.

Imaginal discs can be recovered from cultured embryos mutant for the segment-polarity genes engrailed, naked and patched but not from wingless

Development ◽  
1989 ◽  
Vol 107 (4) ◽  
pp. 715-722 ◽  
Author(s):  
A.A. Simcox ◽  
I.J.H. Roberts ◽  
E. Hersperger ◽  
M.C. Gribbin ◽  
A. Shearn ◽  
...  
Keyword(s):  
Imaginal Discs ◽  
Larval Cuticle ◽  
Segment Polarity Genes ◽  
Abnormal Morphology ◽  
In Vivo Culture ◽  
Segment Polarity ◽  
Abnormal Pattern ◽  
Drosophila Embryos

Drosophila embryos homozygous for strong mutations in each of the segment-polarity genes wingless (wg), engrailed (en), naked (nkd) and patched (ptc) form a larval cuticle in which there is a deletion in every segment. The mutant embryos normally fail to hatch but by in vivo culture we were able to show which could produce adult structures. Cultured wg- embryos did not produce any adult structures. Cultured en- embryos produced eye-antennal derivatives and rarely produced partial thoracic structures. nkd- and ptc- embryos produced eye-antennal and thoracic derivatives. The nkd- and ptc- thoracic imaginal discs developed with an abnormal morphology and abnormal pattern of en- expression. Our findings are consistent with the idea that the thoracic imaginal discs derive from two adjacent groups of cells that express wg and en respectively in the embryo.

scholarly journals Spatial patterns of ecdysteroid receptor activation during the onset ofDrosophilametamorphosis

Development ◽  
2002 ◽  
Vol 129 (7) ◽  
pp. 1739-1750 ◽  
Author(s):  
Tatiana Kozlova ◽  
Carl S. Thummel
Keyword(s):  
Spatial Patterns ◽  
Transgenic Animals ◽  
Receptor Activation ◽  
Imaginal Discs ◽  
Binding Domain ◽  
Dominant Negative ◽  
Ecdysteroid Receptor ◽  
No Activation ◽  
Third Instar Larvae

Ecdysteroid signaling in insects is transduced by a heterodimer of the EcR and USP nuclear receptors. In order to monitor the temporal and spatial patterns of ecdysteroid signaling in vivo we established transgenic animals that express a fusion of the GAL4 DNA binding domain and the ligand binding domain (LBD) of EcR or USP, combined with a GAL4-dependent lacZ reporter gene. The patterns of β-galactosidase expression in these animals indicate where and when the GAL4-LBD fusion protein has been activated by its ligand in vivo. We show that the patterns of GAL4-EcR and GAL4-USP activation at the onset of metamorphosis reflect what would be predicted for ecdysteroid activation of the EcR/USP heterodimer. No activation is seen in mid-third instar larvae when the ecdysteroid titer is low, and strong widespread activation is observed at the end of the instar when the ecdysteroid titer is high. In addition, both GAL4-EcR and GAL4-USP are activated in larval organs cultured with 20-hydroxyecdysone (20E), consistent with EcR/USP acting as a 20E receptor. We also show that GAL4-USP activation depends on EcR, suggesting that USP requires its heterodimer partner to function as an activator in vivo. Interestingly, we observe no GAL4-LBD activation in the imaginal discs and ring glands of late third instar larvae. Addition of 20E to cultured mid-third instar imaginal discs results in GAL4-USP activation, but this response is not seen in imaginal discs cultured from late third instar larvae, suggesting that EcR/USP loses its ability to function as an efficient activator in this tissue. We conclude that EcR/USP activation by the systemic ecdysteroid signal may be spatially restricted in vivo. Finally, we show that GAL4-EcR functions as a potent and specific dominant negative at the onset of metamorphosis, providing a new tool for characterizing ecdysteroid signaling pathways during development.

scholarly journals PRESENCE OF A BASE LINE LEVEL OF SISTER CHROMATID EXCHANGES IN SOMATIC CELLS OF DROSOPHILA MELANOGASTER IN VIVO AND IN VITRO

Genetics ◽  
1982 ◽  
Vol 100 (2) ◽  
pp. 259-278
Author(s):  
Hideo Tsuji
Keyword(s):  
Drosophila Melanogaster ◽  
Sister Chromatid ◽  
Ganglion Cells ◽  
Sister Chromatid Exchanges ◽  
Base Line ◽  
Cell Cycles ◽  
Chromatid Exchanges ◽  
Third Instar Larvae

ABSTRACT Sister chromatid exchanges (SCEs) under in vivo and in vitro conditions were examined in ganglion cells of third-instar larvae of Drosophila melanogaster (Oregon-R). In the in vivo experiment, third-instar larvae were fed on synthetic media containing 5-bromo-2′-deoxyuridine (BrdUrd). After two cell cycles, ganglia were dissected and treated with colchicine. In the in vitro experiment, the ganglia were also incubated in media containing BrdUrd for two cell cycles, and treated with colchicine. SCEs were scored in metaphase stained with Hoechst 33258 plus Giemsa. The frequencies of SCEs stayed constant in the range of 25-150 vg/ml and 0.25-2.5 vg/ml of BrdUrd in vivo and in vitro, respectively. SCEs gradually increased at higher concentrations, strongly suggesting that at least a fraction of the detected SCEs are spontaneous. The constant levels of SCE frequency were estimated, on the average, at 0.103 per cell per two cell cycles for females and 0.101 for males in vivo and at 0.096 for females and 0.091 for males in vitro. No difference was found in the SCE frequency between sexes at any of the BrdUrd concentrations. The analysis for the distribution of SCEs within chromosomes revealed an extraordinarily high proportion of the SCEs at the junctions between euchromatin and heterochromatin; the remaining SCEs were preferentially localized in the euchromatic regions of the chromosomes and in the heterochromatic Y chromosome. These results were largely inconsistent with those of Gatti et al. (1979).

LATTICE SIMULATION OF NASCENT PEPTIDE FOLDING

2004 ◽  
Vol 18 (15) ◽  
pp. 2195-2202 ◽  
Author(s):  
JIAFENG ZHUO ◽  
LINSEN ZHANG ◽  
CHANGJUN CHEN ◽  
YI HE ◽  
YI XIAO
Keyword(s):  
Lattice Model ◽  
Peptide Folding ◽  
Synthesis Process ◽  
Lattice Simulation ◽  
Native State ◽  
Second Stage ◽  
Nascent Peptide ◽  
Two Stages

The nascent peptide folding in vivo is different from the denatured peptide refolding in vitro and can be divided into two stages. In the first stage, the peptide is folding as it is being synthesized until the whole peptide chain is synthesized. The final conformation formed in this stage is called as nascent state. In the second stage, the protein folds beginning with the nascent state formed in the first stage into the native state. We use a lattice model to simulate these two stages and investigate the folding time of the nascent peptide comparing with that of the denatured peptide refolding. Our results show that the synthesis process may affect the folding time of the nascent peptide. This may be helpful to understand why the former folds faster than the latter.

scholarly journals A Mathematical Model for Thermosensitive Liposomal Delivery of Doxorubicin to Solid Tumour

2013 ◽  
Vol 2013 ◽  
pp. 1-13 ◽  
Author(s):  
Wenbo Zhan ◽  
Xiao Yun Xu
Keyword(s):  
Drug Delivery ◽  
Side Effects ◽  
Solid Tumour ◽  
Transport Processes ◽  
Direct Infusion ◽  
Biochemical Processes ◽  
Normal Tissues ◽  
Thermosensitive Liposome ◽  
Anticancer Treatment

The effectiveness of anticancer treatments is often hampered by the serious side effects owing to toxicity of anticancer drugs and their undesirable uptake by healthy cells in vivo. Thermosensitive liposome-mediated drug delivery has been developed as part of research efforts aimed at improving therapeutic efficacy while reducing the associated side effect. Since multiple steps are involved in the transport of drug-loaded liposomes, drug release, and its uptake, mathematical models become an indispensible tool to analyse the transport processes and predict the outcome of anticancer treatment. In this study, a computational model is developed which incorporates the key physical and biochemical processes involved in drug delivery and cellular uptake. The model has been applied to idealized tumour geometry, and comparisons are made between continuous infusion of doxorubicin and thermosensitive liposome-mediated delivery. Results show that thermosensitive liposome-mediated delivery performs better in reducing drug concentration in normal tissues, which may help lower the risk of associated side effects. Compared with direct infusion over a 2-hour period, thermosensitive liposome delivery leads to a much higher peak intracellular concentration of doxorubicin, which may increase cell killing in tumour thereby enhancing the therapeutic effect of the drug.

CD200 is a novel p53-target gene involved in apoptosis-associated immune tolerance

Blood ◽  
2004 ◽  
Vol 103 (7) ◽  
pp. 2691-2698 ◽  
Author(s):  
Michael D. Rosenblum ◽  
Edit Olasz ◽  
Jeffery E. Woodliff ◽  
Bryon D. Johnson ◽  
Marja C. Konkol ◽  
...  
Keyword(s):  
Target Gene ◽  
Molecular Mechanisms ◽  
Apoptotic Cell ◽  
Immune Reactivity ◽  
Biochemical Processes ◽  
The Face ◽  
P53 Target Gene ◽  
Self Antigens

Abstract During apoptotic cell death, biochemical processes modify self-proteins and create potential autoantigens. To maintain self-tolerance in the face of natural cell turnover, the immune system must prevent or control responses to apoptosis-associated autoantigens or risk autoimmunity. The molecular mechanisms governing this process remain largely unknown. Here, we show that expression of the immunoregulatory protein CD200 increases as murine dendritic cells (DCs) undergo apoptosis. We define CD200 as a p53-target gene and identify both p53- and caspase-dependent pathways that control CD200 expression during apoptosis. CD200 expression on apoptotic DCs diminishes proinflammatory cytokine production in response to self-antigens in vitro and is required for UVB-mediated tolerance to haptenated self-proteins in vivo. Up-regulation of CD200 may represent a novel mechanism, whereby immune reactivity to apoptosis-associated self-antigens is suppressed under steady state conditions. (Blood. 2004;103: 2691-2698)

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