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General Information
Symbol
Dmel\EcRB1-ΔC655.W650A.UAS
Species
D. melanogaster
Name
FlyBase ID
FBal0141828
Feature type
allele
Associated gene
Associated Insertion(s)
Carried in Construct
Also Known As
UAS-EcRDN, UAS-EcR-B1W650A, UAS-EcR-DN, UAS-EcR-W650A, EcR-DN, EcRDN, EcRW650A, UAS-EcRW650A, EcRDN, P{UAS-EcR.B1-DeltaC655.W650A}
Nature of the Allele
Mutations Mapped to the Genome
 
Type
Location
Additional Notes
References
Associated Sequence Data
DNA sequence
Protein sequence
 
 
Progenitor genotype
Carried in construct
Cytology
Nature of the lesion
Statement
Reference

UASt regulatory sequences drive expression of a mutated EcR B1 isoform which ends at EcR amino acid residue 655 and has the amino acid replacement W650A.

Allele components
Product class / Tool use(s)
Encoded product / tool
Expression Data
Reporter Expression
Additional Information
Statement
Reference
 
Marker for
Reflects expression of
Reporter construct used in assay
Human Disease Associations
Disease Ontology (DO) Annotations
Models Based on Experimental Evidence ( 0 )
Disease
Evidence
References
Modifiers Based on Experimental Evidence ( 0 )
Disease
Interaction
References
Comments on Models/Modifiers Based on Experimental Evidence ( 0 )
 
Phenotypic Data
Phenotypic Class
Phenotype Manifest In

mesothoracic ventral neurosecretory neuron & axon, with Scer\GAL4FMRFa.PS

metathoracic ventral thoracic neurosecretory neuron & axon, with Scer\GAL4FMRFa.PS

microtubule & dorsal multidendritic neuron ddaC | pupal stage, with Scer\GAL4ppk.PG

neuron & ventral nerve cord, with Scer\GAL4Crz.PC

prothoracic ventral neurosecretory neuron & axon, with Scer\GAL4FMRFa.PS

Detailed Description
Statement
Reference

Expressing EcRB1-ΔC655.W650A.UAS under the control of Scer\GAL4CCAP.PP leads to the progressive loss of most CCAP neurons starting from late 2nd instar through to wandering third instar larval stage. This results in a severely defective pupal development, characterized by a failure in head eversion and shortened wings and legs.

Expressing EcRB1-ΔC655.W650A.UAS under the control of Scer\GAL4Crz.PU completely suppresses the death of vCrz peptidergic neurons at 6-7 hours after puparium formation.

Expressing EcRB1-ΔC655.W650A.UAS under the control of Scer\GAL4A9 results in the formation of melanotic spots in the digestive system of dying larvae.

Expressing EcRB1-ΔC655.W650A.UAS under the control of Scer\GAL4Burs.PL results in the loss of most CCAP neurons in third instar larvae.

Expression of EcRB1-ΔC655.W650A.Scer\UAS in astrocytes, under the control of Scer\GAL4alrm.PD, does not affect astrocyte morphology in 3rd instar larvae. However, EcRB1-ΔC655.W650A.Scer\UAS expression completely blocks their morphological transformation into the highly vacuolated morphology apparent in controls at 6 hours after pupal formation.

Astrocyte morphological transformation can be suppressed when EcRB1-ΔC655.W650A.Scer\UAS is expressed in one to two astrocytes, using MARCM, rather than the entire population. EcRB1-ΔC655.W650A.Scer\UAS expression blocks the induction of phagolysomal activity.

Blockage of EcR signaling in astrocytes, through expression of EcRB1-ΔC655.W650A.Scer\UAS under the control of Scer\GAL4alrm.PD, suppresses the clearance of brp[+] synapses from multiple brain regions at 18 hours after pupal formation and vCrz[+] neuronal debris, with neurite debris lingering until 18 hours after pupal formation.

Astrocytic EcRB1-ΔC655.W650A.Scer\UAS expression, under the control of Scer\GAL4alrm.PD, blocks the transformation of these astrocytes into phagocytes.

While the dorsal and medial lobes of mushroom body γ neurons are largely pruned by 18 hours after pupal formation in controls, pruning of these axonal branches is strongly inhibited by astrocytic EcRB1-ΔC655.W650A.Scer\UAS expression.

In comparison to wild-type white prepupae, fewer and smaller autolysosomes are present in the fat body cells of flies expressing EcRB1-ΔC655.W650A.Scer\UAS under the control of Scer\GAL4Lsp2.PH.

Autophagy is significantly decreased by EcRB1-ΔC655.W650A.Scer\UAS overexpression.

Expression of EcRB1-ΔC655.W650A.Scer\UAS in the remodelling fat body, under the control of Scer\GAL4Lsp2.PH delays pupal development by more than 8 hours.

Severing of the dendrites of ddaC neurons occurs in animals expressing EcRB1-ΔC655.W650A.Scer\UAS under the control of Scer\GAL4repo does occur, albeit with some delay. However, the glial membranes that wrap the proximal dendrites as part of this dendrite pruning process show very little sign of retraction or disintegration by 16 hours after puparium formation and the wrapped dendrite segments remain intact. This defect persist until the death of the neurons during metamorphosis.

Expression of EcRB1-ΔC655.W650A.Scer\UAS under the control of Scer\GAL4twi.PG results in embryos with arrested midgut morphogenesis (midgut constriction and folding is impeded), while tracheal development, head involution and dorsal closure are unaffected.

Expression of EcRB1-ΔC655.W650A.Scer\UAS under the control of Scer\GAL4btl.PS at 20[o]C results in embryos with bloated dorsal tracheal trunks. If the temperature is raised to 29[o]C at the time of the 20-hydroxy-ecdysone pulse, the phenotype is more severe, with impaired tracheal branch growth.

Embryos expressing EcRB1-ΔC655.W650A.Scer\UAS under the control of Scer\GAL469B show incomplete head involution and dorsal closure, as well as a reduced head skeleton and denticle belts.

Follicular cells expressing EcRB1-ΔC655.W650A.Scer\UAS under the control of Scer\GAL4e22c have normal cell shape at S10A. Cells are smaller than normal by S10B, and no apical constriction occurs. Cell nuclei are slightly smaller compared to control cells.

Follicular cells expressing EcRB1-ΔC655.W650A.Scer\UAS under the control of Scer\GAL4e22c block stretch cell formation.

Neuroblasts are found 24hr after puparium formation in 18 out of 101 MARCM clones expressing EcRB1-ΔC655.W650A.Scer\UAS, in contrast to controls which show no neuroblasts at this stage.

Motorneuron clones of a laterally located lineage (identified as lineage 15) expressing EcRB1-ΔC655.W650A.Scer\UAS show abnormally clumped and reduced dendritic arbors that leave large spaces devoid of dendrites. The axons of these clones generally show missing arbors in the adult femur and tibia, misaligned and clumped neuromuscular junctions, and about half the number of femoral endplates of controls. In all seven clones examined, proximal femoral branches were clumped or misaligned showing a star-shaped morphology projecting away from the primary tract. In approximately half the clones examined, the distal femoral arbors are completely missing, but of those still present most are misaligned. In a second, more medially located neuroblast clone (identified as lineage 7), similar phenotypes are seen with clumping and severe reduction in the extent of the dendritic arbor.

In the larval first thoracic neuromere, 9 of 11 clones of the neuroblast lineage identified as lineage 7, expressing EcRB1-ΔC655.W650A.Scer\UAS, the ipsilateral arbors of the secondary projections clump together to produce dense strands interspersed with areas containing no bundles. The midline crossing primary bundle also defasciculated in five out of the 11 clones, and the contralateral arbor shows clumping. Lineage 7 neuroblast clones of the larval third thoracic neuromere overexpressing EcRB1-ΔC655.W650A.Scer\UAS shows some abnormal fasciculation of ipsilateral and contralateral arbors.

Of five clones of lineage 6 neuroblast in the first or the third larval thoracic neuromere, expressing EcRB1-ΔC655.W650A.Scer\UAS, one had a clumped ipsilateral arbor, and two showed dorsal projecting neurons with both reduced and clumped ipsilateral arbors.

Three of four dorsally located lineage 18 neuroblast clones expressing EcRB1-ΔC655.W650A.Scer\UAS show a reduced ipsilateral arbor, with two of these showing clumping together of the terminal branching.

Seven of fourteen clones of lineage 9 neuroblast clones expressing expressing EcRB1-ΔC655.W650A.Scer\UAS show defasciculation of the primary bundle as it curves ventrally around the leg neuropil, compared to one out of six control clones.

Expression of EcRB1-ΔC655.W650A.Scer\UAS in ddaC neurons under the control of Scer\GAL4109(2)80 restricts ddaC dendritic pruning resulting in excess primary and secondary dendrites remaining attached to soma at 18h APF.

There is no sign of microtubule disruption in the dendrites of ddaC neurons at 6 hours after puparium formation (APF) in animals expressing EcRB1-ΔC655.W650A.Scer\UAS under the control of Scer\GAL4ppk.PG (microtubules in the dendrites of wild-type ddaC neurons show breakage at this stage).

Scer\GAL4da.G32-mediated expression of EcRB1-ΔC655.W650A.Scer\UAS results in a mild persistent amnioserosa phenotype. Development of the midgut is also inhibited, but a coherent amnioserosa tissue is evident in innervated and contractile embryos, suggesting amnioserosa degeneration is indeed delayed.

Expression of EcRB1-ΔC655.W650A.Scer\UAS under the control of Scer\GAL4da.Switch.PT in the presence of 1μg ml[-1] RU486 in male flies results in a 10% increase in the median lifespan compared to controls. Higher levels of EcRB1-ΔC655.W650A.Scer\UAS expression (using increasing concentrations of RU486) abolish this positive effect and even result in a decrease in median lifespan (14% decreased compared to controls at a concentration of 50μg ml[-1] RU486).

Expression of EcRB1-ΔC655.W650A.Scer\UAS under the control of Scer\GAL4da.Switch.PT in the presence of RU486 in female flies decreases lifespan in a RU486 dose-dependent manner.

Expression of EcRB1-ΔC655.W650A.Scer\UAS under the control of Scer\GAL4ppk.1.9 results in a significant reduction in the mean density of the dendrites of the ddaC dendritic arborisation neurons compared to controls. The higher order dendritic branches are primarily affected.

Targeted expression of EcRB1-ΔC655.W650A.Scer\UAS driven by Scer\GAL4αTub84B.PP abrogates metamorphosis of the CSD interneuron (CSDn), resulting in a 'larva-like' morphology.

Thoracic ventral neurosecretory cells that express EcRB1-ΔC655.W650A.Scer\UAS under the control of Scer\GAL4Fmrf.PS are of normal size and larval morphology at the start of metamorphosis. However, these mutants show defective neuronal remodelling during metamorphosis. The axons show much lower levels of pruning compared to wild-type cells and show no filopodia during the pruning phase. Scer\GAL4Fmrf.PS>EcRB1-ΔC655.W650A.Scer\UAS neurons fail to extend filopodia during the outgrowth phase. The rare filopodial-like structures that form are very short, stubby and less active than those seen on control cells. Effects on adult axon outgrowth are severe, with arbors significantly reduced in size.

Expression of EcRB1-ΔC655.W650A.Scer\UAS under the control of Scer\GAL4Crz blocks programmed cell death in Crz-expressing ventral nerve cord neurons up to 12 hours after pupal formation.

When expression is governed by Scer\GAL4Lsp2.PH in the larval fat body early autophagosomal structures are able to form but do not acidify, perhaps due to a failure to fuse with lysosomes. At the wandering third instar stage the cytoplasm lacks the usual autophagosomes and large 2-5 micron autolysosomes, and instead contain many small structures - probably autolysosome related. Autophagic area is markedly reduced. Autophagy has been inhibited. In clones, smaller and fewer acidic structures are evident than in wild type, and fat droplets appear distinct.

External Data
Interactions
Show genetic interaction network for Enhancers & Suppressors
Phenotypic Class
Suppressed by
Statement
Reference
NOT suppressed by
Phenotype Manifest In
Suppressed by
Statement
Reference
NOT suppressed by
Additional Comments
Genetic Interactions
Statement
Reference

Overexpression of drprI.Scer\UAS (under the control of Scer\GAL4repo) in a EcRB1-ΔC655.W650A.Scer\UAS background fails to transform astrocytes into phagocytes at puparium formation.

Co-expression of Sox14fl.Scer\UAS with EcRB1-ΔC655.W650A.Scer\UAS in ddaC neurons under the control of Scer\GAL4109(2)80 suppresses the excess dendritic arborization found upon expression of EcRB1-ΔC655.W650A.Scer\UAS alone in ddaC neurons under the control of Scer\GAL4109(2)80.

Co-expression of MicalScer\UAS.fl with EcRB1-ΔC655.W650A.Scer\UAS in ddaC neurons under the control of Scer\GAL4109(2)80 partially suppresses the excess dendritic arborization found upon expression of EcRB1-ΔC655.W650A.Scer\UAS alone in ddaC neurons under the control of Scer\GAL4109(2)80.

Co-expression of MicalN-ter.Scer\UAS with EcRB1-ΔC655.W650A.Scer\UAS in ddaC neurons under the control of Scer\GAL4109(2)80 fails to affect the excess dendritic arborization found upon expression of EcRB1-ΔC655.W650A.Scer\UAS alone in ddaC neurons under the control of Scer\GAL4109(2)80.

The deleterious effect on female lifespan of expression of EcRB1-ΔC655.W650A.Scer\UAS under the control of Scer\GAL4da.Switch.PT in the presence of RU486 is suppressed by ovoD1.

Xenogenetic Interactions
Statement
Reference
Complementation and Rescue Data
Partially rescued by
Comments

Expressing either EcRB1.UAS or EcRB1.UAS under the control of Scer\GAL4CCAP.PP fully rescues CCAP neuronal death in larvae and the pupal phenotype (failure in head eversion and shortened wings and legs) induced by expression of EcRB1-ΔC655.W650A.UAS alone. Expressing EcRA.UAS results only in a partial rescue of both larval CCAP neuronal death and the reduced size of pupal legs and wings, accompanied by a full rescue of the head eversion phenotype.

The defects in midgut morphogenesis which are seen in embryos expressing EcRB1-ΔC655.W650A.Scer\UAS under the control of Scer\GAL4twi.PG are rescued by co-expression of EcRA.Scer\UAS.

The defects in tracheal morphogenesis which are seen in embryos expressing EcRB1-ΔC655.W650A.Scer\UAS under the control of Scer\GAL4btl.PS are rescued by co-expression of EcRA.Scer\UAS.

The embryonic defects caused by expression or EcRB1-ΔC655.W650A.Scer\UAS under the control of Scer\GAL469B are rescued by co-expression of EcRA.Scer\UAS.

Images (0)
Mutant
Wild-type
Stocks (1)
Notes on Origin
Discoverer
Comments
Comments

Encodes a dominant negative form of EcR.

External Crossreferences and Linkouts ( 0 )
Synonyms and Secondary IDs (4)
Reported As
Symbol Synonym
EcRB1-ΔC655.W650A.Scer\UAS
EcRB1-ΔC655.W650A.UAS
Name Synonyms
Secondary FlyBase IDs
    References (42)