puparium formation
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Genetics ◽  
2020 ◽  
Vol 216 (4) ◽  
pp. 1137-1152
Author(s):  
Clinton Rice ◽  
Stuart J. Macdonald ◽  
Xiaochen Wang ◽  
Robert E. Ward

Imaginal disc morphogenesis during metamorphosis in Drosophila melanogaster provides an excellent model to uncover molecular mechanisms by which hormonal signals effect physical changes during development. The broad (br) Z2 isoform encodes a transcription factor required for disc morphogenesis in response to 20-hydroxyecdysone, yet how it accomplishes this remains largely unknown. Here, we use functional studies of amorphic br5 mutants and a transcriptional target approach to identify processes driven by br and its regulatory targets in leg imaginal discs. br5 mutants fail to properly remodel their basal extracellular matrix (ECM) between 4 and 7 hr after puparium formation. Additionally, br5 mutant discs do not undergo the cell shape changes necessary for leg elongation and fail to elongate normally when exposed to the protease trypsin. RNA-sequencing of wild-type and br5 mutant leg discs identified 717 genes differentially regulated by br, including a large number of genes involved in glycolysis, and genes that encode proteins that interact with the ECM. RNA interference-based functional studies reveal that several of these genes are required for adult leg formation, particularly those involved in remodeling the ECM. Additionally, br Z2 expression is abruptly shut down at the onset of metamorphosis, and expressing it beyond this time results in failure of leg development during the late prepupal and pupal stages. Taken together, our results suggest that br Z2 is required to drive ECM remodeling, change cell shape, and maintain metabolic activity through the midprepupal stage, but must be switched off to allow expression of pupation genes.


2007 ◽  
Vol 176 (6) ◽  
pp. 737-739 ◽  
Author(s):  
Carl S. Thummel

The precise determination of when and where cells undergo programmed cell death is critical for normal development and tissue homeostasis. Cao et al. (2007; see p. 843 of this issue) report that the Fork head (Fkh) transcription factor, which is essential for the early development and function of the larval salivary glands in Drosophila melanogaster, also contributes to its demise. These authors show that fkh expression in the salivary glands is normally lost at puparium formation, which is ∼12 h before they undergo massive cell death triggered by the steroid hormone ecdysone, making room for their developing adult counterparts. The loss of Fkh eliminates its role in blocking cell death, allowing for subsequent ecdysone-induced reaper and head involution defective death activator expression and tissue destruction. This study provides new insights into the transcriptional regulation of programmed cell death and the mechanisms that underlie the precise spatial and temporal control of hormone responses during development.


2006 ◽  
Vol 295 (1) ◽  
pp. 156-163 ◽  
Author(s):  
Christoph Scherfer ◽  
Mousumi R. Qazi ◽  
Kuniaki Takahashi ◽  
Ryu Ueda ◽  
Mitchell S. Dushay ◽  
...  

Development ◽  
2000 ◽  
Vol 127 (23) ◽  
pp. 5203-5212
Author(s):  
K. Jacobs ◽  
M.G. Todman ◽  
M.J. Allen ◽  
J.A. Davies ◽  
J.P. Bacon

The tergotrochanteral (jump) motorneuron is a major synaptic target of the Giant Fibre in Drosophila. These two neurons are major components of the fly's Giant-Fibre escape system. Our previous work has described the development of the Giant Fibre in early metamorphosis and the involvement of the shaking-B locus in the formation of its electrical synapses. In the present study, we have investigated the development of the tergotrochanteral motorneuron and its electrical synapses by transforming Drosophila with a Gal4 fusion construct containing sequences largely upstream of, but including, the shaking-B(lethal) promoter. This construct drives reporter gene expression in the tergotrochanteral motorneuron and some other neurons. Expression of green fluorescent protein in the motorneuron allows visualization of its cell body and its subsequent intracellular staining with Lucifer Yellow. These preparations provide high-resolution data on motorneuron morphogenesis during the first half of pupal development. Dye-coupling reveals onset of gap-junction formation between the tergotrochanteral motorneuron and other neurons of the Giant-Fibre System. The medial dendrite of the tergotrochanteral motorneuron becomes dye-coupled to the peripheral synapsing interneurons between 28 and 32 hours after puparium formation. Dye-coupling between tergotrochanteral motorneuron and Giant Fibre is first seen at 42 hours after puparium formation. All dye coupling is abolished in a shaking-B(neural) mutant. To investigate any interactions between the Giant Fibre and the tergotroachanteral motorneuron, we arrested the growth of the motorneuron's medial neurite by targeted expression of a constitutively active form of Dcdc42. This results in the Giant Fibre remaining stranded at the midline, unable to make its characteristic bend. We conclude that Giant Fibre morphogenesis normally relies on fasciculation with its major motorneuronal target.


Development ◽  
2000 ◽  
Vol 127 (1) ◽  
pp. 97-107 ◽  
Author(s):  
F. Roch ◽  
M. Akam

The Drosophila haltere is a much reduced and specialised hind wing, which functions as a balance organ. Ultrabithorax (Ubx) is the sole Hox gene responsible for the differential development of the fore-wing and haltere in Drosophila. Previous work on the downstream effects of Ubx has focused on the control of pattern formation. Here we provide the first detailed description of cell differentiation in the haltere epidermis, and of the developmental processes that distinguish wing and haltere cells. By the end of pupal development, haltere cells are 8-fold smaller in apical surface area than wing cells; they differ in cell outline, and in the size and number of cuticular hairs secreted by each cell. Wing cells secrete only a thin cuticle, and undergo apoptosis within 2 hours of eclosion. Haltere cells continue to secrete cuticle after eclosion. Differences in the shape of wing and haltere cells reflect differences in the architecture of the actin cytoskeleton that become apparent between 24 and 48 hours after puparium formation. We show that Ubx protein is not needed later than 6 hours after puparium formation to specify these differences, though it is required at later stages for the correct development of campaniform sensilla on the haltere. We conclude that, during normal development, Ubx protein expressed before pupation controls a cascade of downstream effects that control changes in cell morphology 24–48 hours later. Ectopic expression of Ubx in the pupal wing, up to 30 hours after puparium formation, can still elicit many aspects of haltere cell morphology. The response of wing cells to Ubx at this time is sensitive to both the duration and level of Ubx exposure.


Genome ◽  
1999 ◽  
Vol 42 (5) ◽  
pp. 919-929 ◽  
Author(s):  
Angeliki Gariou-Papalexiou ◽  
George Chrysanthis ◽  
Anastassios C Mintzas ◽  
Antigone Zacharopoulou

The patterns of puffing activity in the salivary gland polytene chromosomes have been studied during the late larval and prepupal stages of the medfly Ceratitis capitata. A total of 128 loci, with significant changes in puffing activity during this developmental period, were assigned to the five autosomes of the medfly. Two waves of puffing activity were observed, the first during the late larval stage and the second during the prepupal development. Overall puffing activity can be divided into four groups, group-IV activity being most conspicuous with 58 active loci. The major changes in puffing activity take place around jumping, a characteristic event occurring about 6 h before puparium formation, at puparium formation, and during midprepupal development. The overall puffing activity shows a positive correlation to the ecdysone titer in the hemolymph, suggesting that most of the changes in the activity of the puffs during the late larval and prepupal stages of the medfly may be regulated by ecdysone.Key words: polytene chromosomes, puffing patterns, ecdysone, Ceratitis capitata.


Development ◽  
1999 ◽  
Vol 126 (18) ◽  
pp. 4065-4076 ◽  
Author(s):  
T. Lee ◽  
A. Lee ◽  
L. Luo

The mushroom bodies (MBs) are prominent structures in the Drosophila brain that are essential for olfactory learning and memory. Characterization of the development and projection patterns of individual MB neurons will be important for elucidating their functions. Using mosaic analysis with a repressible cell marker (Lee, T. and Luo, L. (1999) Neuron 22, 451–461), we have positively marked the axons and dendrites of multicellular and single-cell mushroom body clones at specific developmental stages. Systematic clonal analysis demonstrates that a single mushroom body neuroblast sequentially generates at least three types of morphologically distinct neurons. Neurons projecting into the (gamma) lobe of the adult MB are born first, prior to the mid-3rd instar larval stage. Neurons projecting into the alpha' and beta' lobes are born between the mid-3rd instar larval stage and puparium formation. Finally, neurons projecting into the alpha and beta lobes are born after puparium formation. Visualization of individual MB neurons has also revealed how different neurons acquire their characteristic axon projections. During the larval stage, axons of all MB neurons bifurcate into both the dorsal and medial lobes. Shortly after puparium formation, larval MB neurons are selectively pruned according to birthdays. Degeneration of axon branches makes early-born gamma neurons retain only their main processes in the peduncle, which then project into the adult gamma lobe without bifurcation. In contrast, the basic axon projections of the later-born (alpha'/beta') larval neurons are preserved during metamorphosis. This study illustrates the cellular organization of mushroom bodies and the development of different MB neurons at the single cell level. It allows for future studies on the molecular mechanisms of mushroom body development.


Development ◽  
1998 ◽  
Vol 125 (23) ◽  
pp. 4709-4717 ◽  
Author(s):  
B.L. Hall ◽  
C.S. Thummel

Pulses of the steroid hormone ecdysone function as key temporal signals during insect development, coordinating the major postembryonic developmental transitions, including molting and metamorphosis. In vitro studies have demonstrated that the EcR ecdysone receptor requires an RXR heterodimer partner for its activity, encoded by the ultraspiracle (usp) locus. We show here that usp exerts no apparent function in mid-third instar larvae, when a regulatory hierarchy prepares the animal for the onset of metamorphosis. Rather, usp is required in late third instar larvae for appropriate developmental and transcriptional responses to the ecdysone pulse that triggers puparium formation. The imaginal discs in usp mutants begin to evert but do not elongate or differentiate, the larval midgut and salivary glands fail to undergo programmed cell death and the adult midgut fails to form. Consistent with these developmental phenotypes, usp mutants show pleiotropic defects in ecdysone-regulated gene expression at the larval-prepupal transition. usp mutants also recapitulate aspects of a larval molt at puparium formation, forming a supernumerary cuticle. These observations indicate that usp is required for ecdysone receptor activity in vivo, demonstrate that the EcR/USP heterodimer functions in a stage-specific manner during the onset of metamorphosis and implicate a role for usp in the decision to molt or pupariate in response to ecdysone pulses during larval development.


1998 ◽  
Vol 49 (2) ◽  
pp. 146
Author(s):  
A. Moribayashi ◽  
T. Hayashi ◽  
C. Shudo ◽  
H. Kurahashi

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