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PubMedDate not supplied

Commencement of pupal commitment in late penultimate instar and its hormonal control in wing imaginal discs of the silkworm, Bombyx mori.

Pupal commitment of the wing imaginal disc of the silkworm, Bombyx mori, is completed shortly after the final (fifth) larval ecdysis. Pupal commitment was induced by in vitro culture with 20-hydroxyecdysone (20E). Shortly after the head capsule slippage (HCS) that occurs approximately 24 h before the final larval ecdysis, the discs become competent to respond to 20E, indicating that the process of pupal commitment begins in the late penultimate (fourth) instar. The simultaneous presence of methoprene (JHA) with 20E suppressed the pupal commitment at 4 ng/ml for the discs at 12 h after HCS and at 240 ng/ml for the discs at the ecdysis. Thus, the discs rapidly lose their sensitivity to JH at the end of the fourth instar. Day 0 fourth wing discs were not pupally committed by 20E when freshly dissected discs were exposed to 20E. By contrast, exposure to 20E after a pre-culture in a hormone free medium induced the pupal commitment. In those discs, the effective JHA concentration to suppress the 20E effects was 0.1 ng/ml. The present data suggest that pupal commitment proceeds through two stages from a reversible state that begins at around HCS to an irreversible state early in the fifth instar. The loss of sensitivity to JH is the primary impetus to begin the process and 20E is the factor that drives the discs to enter the reversible state.

Animals↗
PubMed2004

Isolation of an imaginal disc growth factor homologue from Pieris rapae and its expression following parasitization by Cotesia rubecula.

Endoparasitoid insects introduce maternal factors into the body of their host at oviposition to suppress cellular defences for the protection of the developing parasitoid. We have shown that transient expression of polydnavirus genes from a hymenopteran parasitoid Cotesia rubecula (CrPDV) is responsible for the inactivation of hemocytes from the lepidopteran host Pieris rapae. Since the observed downregulation of CrPDV genes in infected host tissues is not due to cis-regulatory elements at the CrV1 gene locus, we speculated that the termination of CrPDV gene expression may be due to cellular inactivation caused by the CrV1-mediated immune suppression of infected tissues. To test this assumption, we isolated an imaginal disc growth factor (IDGF) that is expressed in fat body and hemocytes, the target of viral infection and expression of CrPDV genes. Time-course experiments showed that the level of P. rapae IDGF is not affected by parasitization and polydnavirus infection. However, the amount of highly expressed genes, such as storage proteins, arylphorin and lipophorin, are significantly reduced following parasitization.

Amino Acid Sequence↗
PubMed2004

Usefulness of pain drawings in identifying real or imagined pain: accuracy of pain professionals, nonprofessionals, and a decision model.

UNLABELLED: The aim of this study was to determine the accuracy of pain specialists, nonprofessionals, and a decision model in judging whether a pain diagram was marked by a patient with chronic pain or by a healthy volunteer. Two hundred twenty-eight pain drawings were shown in random order to 10 pain medicine physicians, 10 pain medicine fellows, 10 nonphysician specialists, and 10 nonprofessionals. One half of the drawings (n = 114) had been produced by patients treated at a pain center and the other half (n = 114) by healthy individuals who were instructed to mark the diagrams as if they had a pain problem. The nonprofessionals were found to be 51.5% accurate, pain medicine fellows 52.7%, nonphysician specialists 54.3%, and pain medicine physicians 55.2 % accurate at distinguishing drawings by actual pain patients from drawings from volunteers without pain. A decision model was able to achieve 68.9% accuracy in determining which drawings were made by pain patients and which drawings were made by healthy individuals. The results suggest that subjective assessment of pain drawings alone is not useful in determining whether someone has real or imagined pain. A decision model that makes decisions on the basis of the number of highlighted squares on the pain diagram can identify real pain drawings with greater accuracy than humans. PERSPECTIVE: Pain drawings are clinically useful but have limitations. This study illustrates some of the benefits of computerized pain assessment and highlights the importance of not judging patients on the basis of one source of information.

Adolescent↗
PubMed2004

Cerebral activation during hypnotically induced and imagined pain.

The continuing absence of an identifiable physical cause for disorders such as chronic low back pain, atypical facial pain, or fibromyalgia, is a source of ongoing controversy and frustration among pain physicians and researchers. Aberrant cerebral activity is widely believed to be involved in such disorders, but formal demonstration of the brain independently generating painful experiences is lacking. Here we identify brain areas directly involved in the generation of pain using hypnotic suggestion to create an experience of pain in the absence of any noxious stimulus. In contrast with imagined pain, functional magnetic resonance imaging (fMRI) revealed significant changes during this hypnotically induced (HI) pain experience within the thalamus and anterior cingulate (ACC), insula, prefrontal, and parietal cortices. These findings compare well with the activation patterns during pain from nociceptive sources and provide the first direct experimental evidence in humans linking specific neural activity with the immediate generation of a pain experience.

Adult↗
PubMed2006

The UBX-regulated network in the haltere imaginal disc of D. melanogaster.

Hox proteins have been proposed to act at multiple levels within regulatory hierarchies and to directly control the expression of a plethora of target genes. However, for any specific Hox protein or tissue, very few direct in vivo-regulated target genes have been identified. Here, we have identified target genes of the Hox protein Ultrabithorax (UBX), which modifies the genetic regulatory network of the wing to generate the haltere, a modified hindwing. We used whole-genome microarrays and custom arrays including all predicted transcription factors and signaling molecules in the Drosophila melanogaster genome to identify differentially expressed genes in wing and haltere imaginal discs. To elucidate the regulation of selected genes in more detail, we isolated cis-regulatory elements (CREs) for genes that were specifically expressed in either the wing disc or haltere disc. We demonstrate that UBX binds directly to sites in one element, and these sites are critical for activation in the haltere disc. These results indicate that haltere and metathoracic segment morphology is not achieved merely by turning off the wing and mesothoracic development programs, but rather specific genes must also be activated to form these structures. The evolution of haltere morphology involved changes in UBX-regulated target genes, both positive and negative, throughout the wing genetic regulatory network.

Animals↗
PubMedDate not supplied

Emotional processing and outcome of imaginal flooding therapy in Vietnam veterans with chronic posttraumatic stress disorder.

This study examined emotional processing and outcome in 20 Vietnam veterans with chronic posttraumatic stress disorder (PTSD) who underwent imaginal flooding therapy. Results supported the occurrence of emotional processing, as manifest in significant activation, within-session habituation, and partial across-session habituation of physiologic and self-reported process variables. The flooding therapy produced only modest overall improvement, which was statistically significant for avoidance symptomatology measured by the impact of Events Scale (IOES) and number of intrusions per day recorded by the subject in a log. Symptomatic improvement appeared to generalize from a treated to an untreated experience. Heart rate activation during the first flooding session predicted a decrease in daily number of intrusive combat memories across the therapy. Otherwise, there was little association between extent of emotional processing and therapeutic outcome. The results provide limited support for the notion that mobilization of psychophysiologic arousal during exposure therapy predicts improvement.

Adult↗
PubMed2002

Pupal commitment and its hormonal control in wing imaginal discs.

The timing of pupal commitment of the forewing imaginal discs of the silkworm, Bombyx mori, was determined by a transplantation assay using fourth instar larvae. The wing discs were not pupally committed at the time of ecdysis to the fifth instar. Pupal commitment began shortly after the ecdysis and was completed in 14 h. When the discs of newly molted larvae (0-h discs) were cultured in medium containing no hormone, they were pupally committed in 26 h. In vitro exposure of 0-h discs to 20-hydroxyecdysone accelerated the progression of pupal commitment. Methoprene, a juvenile hormone analog (JHA), did not suppress the change in commitment in vitro at physiological concentrations. Thus the wing discs at the time of the molt have lost their sensitivity to JH, and 20E is not a prerequisite for completion of pupal commitment. These results suggest that the change in commitment in the forewing discs may begin before the last larval molt.

Journal Article↗
PubMed1998

The effect of juvenile hormone on the response of the Drosophila imaginal disc cell line Cl 8+ to moulting hormone.

The Drosophila wing imaginal disc cell line Cl 8+ was used to investigate the interaction between juvenile hormone III (JH) and 20-hydroxyecdysone (20HE). Cell cultures were exposed to either or both hormones at a range of concentrations and cell growth was observed. JH was found to ameliorate the effects of 20HE on cell growth, even when added after the cells had been exposed to 20HE for 4 or 24h.

Journal Article↗
PubMed2000

Juvenile hormone and juvenile hormone mimics inhibit proliferation in a lepidopteran imaginal disc cell line.

The action of juvenile hormone (JH) and JH mimics have been examined in vitro by utilizing the imaginal disc-derived cell line, IAL-PID2. We have discovered that the cell line was responsive to JH and a variety of JH mimics. The most consistent response obtained in our studies was inhibition of cell proliferation, in the absence of 20-hydroxyecdysone (20E), which characteristically reduces cell proliferation in its own right in this cell line. JH-I, JH-III, methoprene, fenoxycarb, and farnesol significantly inhibited cell proliferation after 3 days of exposure of the cells in vitro to each of the compounds. Linoleic acid controls had no effect on proliferation in the cultures. The cell proliferation assay demonstrates the JH responsiveness of this cell line, but the concentrations of JH required were high compared to the concentrations of 20E needed for inhibition of proliferation in these cells.

Journal Article↗
PubMed2000

Signaling reaches to new dimensions in Drosophila imaginal discs.

Finding that peripodial cells in wing and eye imaginal discs are essential for the growth and patterning of the separate layer of disc cells now opens the study of interacting cell layers to the powerful developmental genetic techniques with which the Drosophila system is blessed. We can anticipate that future work will identify how such interactions contribute to patterning and how the mechanisms and processes that are involved are conserved in vertebrates. We can also look forward to contributions that this work will make to understand-ing the role of interconnecting cell extensions in such signaling processes. In this minireview, we have noted numerous types of signaling cells in which cellular extensions have been observed. At present, neither the functional nor structural relationship of these related structures is known. It is certainly tempting to suggest that these structures are conduits for signals or that they function as sensors. There is, as yet, no direct experimental evidence for such roles.

Animals↗
PubMed2000

Peripodial cells regulate proliferation and patterning of Drosophila imaginal discs.

Cells employ a diverse array of signaling mechanisms to establish spatial patterns during development. Nowhere is this better understood than in Drosophila, where the limbs and eyes arise from discrete epithelial sacs called imaginal discs. Molecular-genetic analyses of pattern formation have generally treated discs as single epithelial sheets. Anatomically, however, discs comprise a columnar cell monolayer covered by a squamous epithelium known as the peripodial membrane. Here we demonstrate that during development, peripodial cells signal to disc columnar cells via microtubule-based apical extensions. Ablation and targeted gene misexpression experiments demonstrate that peripodial cell signaling contributes to growth control and pattern formation in the eye and wing primordia. These findings challenge the traditional view of discs as monolayers and provide foundational evidence for peripodial cell function in Drosophila appendage development.

Animals↗
PubMed2000

Dpp gradient formation in the Drosophila wing imaginal disc.

The secreted signaling protein Dpp acts as a morphogen to pattern the anterior-posterior axis of the Drosophila wing. Dpp activity is required in all cells of the developing wing imaginal disc, but the ligand gradient that supports this activity has not been characterized. Here we make use of a biologically active form of Dpp tagged with GFP to examine the ligand gradient. Dpp-GFP forms an unstable extracellular gradient that spreads rapidly in the wing disc. The activity gradient visualized by MAD phosphorylation differs in shape from the ligand gradient. The pMAD gradient adjusted to compartment size when this was experimentally altered. These observations suggest that the Dpp activity gradient may be shaped at the level of receptor activation.

Animals↗
PubMed1999

Cytonemes: cellular processes that project to the principal signaling center in Drosophila imaginal discs.

Wing imaginal disc cells in Drosophila develop by using information received from a signaling center associated with the anterior/posterior compartment border. We show here that disc cells have thin, actin-based extensions (cytonemes) that project to this signaling center. Cytonemes can be induced when cells from the lateral flanks of a wing disc are cultured next to cells from the A/P border or next to a source of fibroblast growth factor. Mouse limb bud cells also grow projections during a brief culture period, indicating that cytonemes are an attribute of both vertebrate and invertebrate cells. We suggest that cytonemes may be responsible for some forms of long-range cell-cell communication.

Actins↗
PubMed2004

Moral imagination in tissue engineering research on animal models.

Animal experimentation is an integrated part of tissue engineering research. This paper investigates what scientists conducting such experimentation should reasonably take into consideration from an ethical point of view. It is argued that scientists should use their moral imagination in making fundamental ethical choices, in reflecting on legal regulation, in taking public opinion seriously, and in balancing human benefit and animal harm as expected outcomes of the experiments.

Animal Experimentation↗
PubMed2000

Imagination and reality in the search for the P2Y receptors.

A great body of evidence based on tissue and organ physiology and pharmacology led to the recognition, widespread by about 1990, that there must be cell membrane receptors for extracellular nucleotides to transduce their effects. This evidence was provided by the pioneering work of Geoffrey Burnstock and those who worked with him, or was developed by others starting from that information. This article will review how we could start from that foundation to clone the first known gene for such a receptor, P2Y(1). Some unusual properties of that receptor were revealed. I will consider further the P2Y receptors as a class - its definition, now that many such genes have become known. Imagination and reality have been intertwined in this saga.

Animals↗
PubMed1975

Characterization and purification of messenger RNAs containing poly A in insect imaginal disks.

The presence of a fragment of polyA resistant to both T1 and p ribonucleases in mRNAs extracted from wing imaginal disks of an insect, Pieris brassicae, is reported. Its length was approximatively 150 nucleotides. PolyU sepharose affinity chromatography was subsequently used for purification of these polyA(+)mRNA molecules. Analyses on sucrose gradients showed a good recovery of poly(+)molecules characterized by their size (20-100 S) and a polydisperse pattern. These mRNAlike species represent 2-3 per cent of the total radioactivity incorporated into RNA in 3 hours of labeling. Sequential extractions were carried out to provide cytoplasmic RNA rich fractions (4 degrees C) and nuclear rich fractions (45 degrees C). When assayed for the presence of polyA(+)RNA, molecules extracted by these two sequential methods were found to be very similar in their polyA content.

Animals↗
PubMed2001

Genomic organization and expression of the human tumorous imaginal disc (TID1) gene.

Human Tid-1, the human homologue of the Drosophila tumor suppressor lethal (2) tumorous imaginal discs, l(2) tid gene product, is a member of the DNAJ family of proteins which serve as co-chaperones to Hsp70 proteins. Here we report the cloning and characterization of the genomic structure of the human TID1 gene (hTID1), which is located on chromosome 16p13.3. hTID1 is approximately 34 kb and is composed of 12 exons. Exon sizes vary from 64 to 232 nucleotides, with the exception of exon 12 corresponding to the 3' untranslated region of hTID1, which extends over 1.1 kb. S1 nuclease protection assays and primer extension experiments indicate a putative transcriptional start site 21 nucleotides upstream of the initiating methionine. The presumptive promoter is characterized by the lack of TATA and CAAT motifs, and a high G+C content. The 5' flanking region contains several consensus binding sites for transcription factors that regulate gene expression during tissue and organ development, such as myeloid zinc finger (MZF1), Ikaros 2 and homeodomain proteins, as well as factors implicated in cell growth and survival responses, including AP-1, PEA3, E2F and NF-kB. Three alternatively spliced variants of hTID1 are expressed in a tissue and cell-type specific manner in many of the human tissues examined. The existence of these forms needs to be considered in efforts aimed at identifying mutations in the hTID1 gene.

5' Flanking Region↗
PubMed1997

Sequence, molecular organization and products of the Drosophila virilis homologs of the D. melanogaster nested genes lethal(2) tumorous imaginal discs [1(2)tid] and lethal(2) neighbour of tid [1(2)not].

In this study, we describe the isolation of the Drosophila virilis (Dvir) 6201-bp genomic fragment homologous to a 7047-bp genomic region of D. melanogaster (Dmel) that harbors the nested genes lethal(2) tumorous imaginal discs (l(2)tid), lethal(2) neighbour of tid (l(2)not) and lethal(2) relative of tid (l(2)rot). The isolated fragment, which maps at the cytogenetic position 50A5 on chromosome 5, carries the Dvir homologs of the Dmel genes l(2)tid and l(2)not. In both cases, the interspecific comparison of the determined sequences reveals a high homology regarding the protein coding regions and a high degree of evolutionary divergence concerning the intronic parts of the genes. In the two distantly related species, the particular gene within gene arrangement of the two genes is conserved, namely, Dvir tid is located in the intron of Dvir not, on the non-coding DNA strand. Interestingly, the Dvir homolog of the Dmel l(2)rot gene residing in the l(2)not intron on its coding strand, opposite l(2)tid, is not present in the 6201-bp genomic fragment. The protein predicted from the Dvir tid sequence, Dvir Tid58, exhibits 76.5% identity with the putative Tid56 protein of Dmel. The putative Dvir Not58 protein shows 71% identity with its Dmel homolog Not56. The developmental transcript and protein patterns, as well as the characteristics of the protein products encoded by the genes Dvir tid and Dvir not are similar to those identified for their Dmel homologs.

Amino Acid Sequence↗