eye, with Scer\GAL4GMR.PF
leg, with Scer\GAL4CCAP.PP
Expression of ShEKO.Scer\UAS.T:Avic\GFP-GL in DA neurons under the control of Scer\GAL4ple.PF causes a phase shift in circadian locomotor activity and gradually extends the circadian periodicity.
Expression of ShEKO.Scer\UAS.T:Avic\GFP-GL in Scer\GAL4Ilp2.PR-neurons results in an increase in sleep both during the day and night.
Expression of one, two, or three copies of ShEKO.Scer\UAS.T:Avic\GFP-GL in N[[CCAP]] neurons under the control of Scer\GAL4Ccap.PP induces wing expansion deficits of increasing severity and frequency. Suppression of N[[CCAP]] by three copies of ShEKO.Scer\UAS.T:Avic\GFP-GL completely eliminates both the tonic abdominal contraction that defines the grooming phase and air swallowing. Despite the absence of the expansion phase, however, the duration of the perch selection phase in these animals is indistinguishable from that of control flies.
Flies expressing two copies of ShEKO.Scer\UAS.T:Avic\GFP-GL in N[[CCAP]] neurons under the control of Scer\GAL4Ccap.PP can be divided into two categories: Approximately half resemble flies expressing one copy of ShEKO.Scer\UAS.T:Avic\GFP-GL, which exhibit a robust expansion phase that includes tonic abdominal contraction and air swallowing, while the rest resemble flies expressing three copies of ShEKO.Scer\UAS.T:Avic\GFP-GL, which completely lack an expansion phase.
Expression of two copies of ShEKO.Scer\UAS.T:Avic\GFP-GL under the control of Scer\GAL4futsch-C380 in the presence of Scer\GAL80Cha.PK (to suppress expression in all cholinergic neurons) results in the MN5 neuron showing strongly decreased excitability in response to a current injection into the soma. Resting membrane potential and input resistance of the MN5 neuron are unaffected.
The dendritic structure of the MN5 neuron is altered compared to wild type in animals expressing two copies of ShEKO.Scer\UAS.T:Avic\GFP-GL under the control of Scer\GAL4futsch-C380. Total dendritic length and the the mean length of individual dendritic branches is increased. The number of branch points and maximum branch order are unaffected. The mean distance of all dendritic segments to the origin of the tree and the total dendritic surface are increased.
Flies expressing two copies of ShEKO.Scer\UAS.T:Avic\GFP-GL under the control of Scer\GAL4futsch-C380 in the presence of Scer\GAL80Cha.PK show reduced flight motor performance in a restrained flight assay. The initial and total flight time are significantly decreased compared to wild type.
Larval RP2 motor neurons expressing ShEKO.Scer\UAS.T:Avic\GFP-GL under the control of Scer\GAL4unspecified show no significant change in dendrite volume compared to controls.
Cultured motor neurons expressing ShEKO.Scer\UAS.T:Avic\GFP-GL under the control of Scer\GAL4futsch-C380 (in the presence of Scer\GAL80Cha.PK to suppress expression in cholinergic neurons) show no significant change in neurite length or branch number compared to controls.
Expression of ShEKO.Scer\UAS.T:Avic\GFP-GL by Scer\GAL4P2.4.Pdf results in an increase of sleep but a decrease in sleep latency in the day and night, compared to controls.
Most animal expressing one copy of ShEKO.Scer\UAS.T:Avic\GFP-GL by Scer\GAL4burs.PP show a partially expanded wing phenotype, a small fraction do not expand their wings, and 40% show normal wing expansion. In contrast, most flies expressing three copies of ShEKO.Scer\UAS.T:Avic\GFP-GL fail to expand their wings. These animals also fail to tan properly, retaining their unpigmented juvenile state 3 hours after eclosion.
Flies expressing a single copy of ShEKO.Scer\UAS.T:Avic\GFP-GL under the control of Scer\GAL4Ccap.PP fail to expand their wings when raised at 18[o]C, but expand their wings normally when raised at 31[o]C. Similarly, flies expressing a single copy of ShEKO.Scer\UAS.T:Avic\GFP-GL by Scer\GAL4burs.PP show approximately five-fold higher frequency in wing expansion defects when raised at 18[o]C than when raised at 31[o]C. A fraction of these animals raised at 18[o]C then shifted to 31[o]C from 5 to 6 days after puparium formation show wing expansion defects.
Although adult flies expressing three copies of ShEKO.Scer\UAS.T:Avic\GFP-GL under the control of Scer\GAL4dimm-929 fail to expand their wings, they consistently show a period of sustained abdominal contraction characteristic of the wing expansion phase after eclosion and they ingest air similarly to wild-type flies. However, in contrast to wild-type flies, the abdomen is elongated and constricted, but not necessarily flexed.
Only 4% of adults expressing three copies of ShEKO.Scer\UAS.T:Avic\GFP-GL under the control of Scer\GAL4burs.PP show sustained abnormal abdominal contractions during the wing expansional phase after eclosion, and only 14% swallow air (all mutant animals that swallow air are female).
Expression of ShEKO.Scer\UAS.T:Avic\GFP-GL under the control of Scer\GAL4Mef2.247.Switch (in the presence of RU486) causes a significant increase in sleep.
Expression of increasing copy numbers of ShEKO.Scer\UAS.T:Avic\GFP-GL in the wing under the control of Scer\GAL4Ccap.PP results in incremental increases in the frequency and severity of wing expansion deficits. Most flies expressing 1xShEKO.Scer\UAS.T:Avic\GFP-GL partially expand their wings, whereas all flies expressing 3xShEKO.Scer\UAS.T:Avic\GFP-GL have unexpanded wings. Flies expressing 2xShEKO.Scer\UAS.T:Avic\GFP-GL (under the control of Scer\GAL4Ccap.PP), typically lack the level of pigmentation observed in age-matched controls 3hrs after eclosion. Inhibition of melanisation in these flies appears complete, with the differences in cuticle pigmentation between ShEKO.Scer\UAS.T:Avic\GFP-GL-expressing and control flies comparable with those between newly eclosed and 3hr-old wild-type flies. A small number of animals expressing 3xShEKO.Scer\UAS.T:Avic\GFP-GL (under the control of Scer\GAL4Ccap.PP) die with head eversion defects and foreshortened wings and legs. Pharate adults expressing 3xShEKO.Scer\UAS.T:Avic\GFP-GL (under the control of Scer\GAL4Ccap.PP) exhibit no loss of NCCAP neurons. Expression of ShEKO.Scer\UAS.T:Avic\GFP-GL (under the control of Scer\GAL4dimm-929) incrementally increases the severity of wing expansion deficits with increasing transgene copy numbers, with expression of 3xShEKO.Scer\UAS.T:Avic\GFP-GL resulting in unexpanded wings in 100% of animals and failure to tan. Flies expressing Scer\GAL80Ccap, as well as ShEKO.Scer\UAS.T:Avic\GFP-GL (under the control of Scer\GAL4dimm-929) exhibited wild-type levels of wing expansion.
Flies expressing ShEKO.Scer\UAS.T:Avic\GFP-GL, under the control of Scer\GAL4NP1535, exhibit a significantly lowered proboscis extension reflex rate in response to 100mM sucrose. The gustatory receptor neuron response to 100mM sucrose is reduced in these flies.
Expression of ShEKO.Scer\UAS.T:Avic\GFP-GL in Rh6 photoreceptors, under the control of Scer\GAL4Rh6.PD, does not affect 5-HT arborization. Expression of ShEKO.Scer\UAS.T:Avic\GFP-GL in all photoreceptors, under the control of Scer\GAL4GMR.PF, does not affect 5-HT arborization, but does reduce larval response to light.
When ShEKO.Scer\UAS.T:Avic\GFP-GL is driven by Scer\GAL4elav.PLu, it is embryonic lethal. When ShEKO.Scer\UAS.T:Avic\GFP-GL is driven by Scer\GAL4how-24B, larvae survive until third instar. When ShEKO.Scer\UAS.T:Avic\GFP-GL under the control of Scer\GAL4Mhc.Switch.PO in the presence of RU486, K+ currents on MF6 (in abdominal segments 2-4) are abnormal. Outward K+ currents of about 315nA are seen at a holding potential of +20mV a seven fold increase over controls. When RU486 is not present 36% increase in current is seen over controls.
When ShEKO.Scer\UAS.T:Avic\GFP-GL is driven by Scer\GAL4how-24B the number and gross morphology of the sensory neurons appears unaltered. When ShEKO.Scer\UAS.T:Avic\GFP-GL is driven by Scer\GAL4elav-C155 or Scer\GAL4how-24B K+ current densities in embryonic neurons and muscles respectively are over 2-fold larger than those of controls. Tha activation threshold of these currents is also substantially shifted in the hyperpolarising direction in both cell types: -30 mV in neurons and -26 in muscles. Expression of ShEKO.Scer\UAS.T:Avic\GFP-GL significantly reduces cellular excitability and indeed no muscle contractions are seen in mutant muscle fibres. Decreases in both input resistances and resting membrane potentials are seen in muscles. On average, resting potentials are hyperpolarised by -9mV in mutant muscles and input resistances measured at the resting potential are 74% smaller. When synaptic function in muscle fibre is measured in response to retrograde stimulation of axon 2 in the segmental nerve, while excitatory postsynaptic currents (EPSCs) are similar to controls excitatory postsynaptic potentials (EPSPs) are on average 30% smaller and decay nearly five times as fast. Expression of one copy of ShEKO.Scer\UAS.T:Avic\GFP-GL driven by either Scer\GAL4elav-C155 or Scer\GAL4how-24B leads to increased mortality. Two copies are lethal. If two copies of are driven by Scer\GAL4elav-C155, 99% of embryos fail to hatch. When driven by Scer\GAL4how-24B lethality is primarily in larval and pupal stages. This lethality is dosage dependant. In both nerves and muscles, the increasing lethality with increasing dosage correlates with progressive impairment to motor functions. Mutant embryos with the Scer\GAL4elav-C155 driver show a reduction in the frequency of full-body peristaltic waves. When the VLM innervation pattern of mutant embryos with the Scer\GAL4elav-C155 driver are examined at late stage 17, just after developmental arrest, abnormalities are seen. There is an approximately 100% increase in both the frequency and average length of the collateral branches. No defects are seen in these ectopic synapses. When ShEKO.Scer\UAS.T:Avic\GFP-GL is driven by Scer\GAL4GMR.PF, defects are seen in the eye's response to a light stimulus. the initial amplitude of the photoreceptor potential declines progressively with increasing transgene copy number, with suppression saturating at approximately 50% of controls. In contrast, the postsynaptic response of the laminar interneurons remains relatively invariant. When three copies of the transgene responsible for ShEKO.Scer\UAS.T:Avic\GFP-GL are driven by Scer\GAL4A307 various anatomical and behavioural defects are seen. There is pronounced pigmentation of the thoracic trident, a failure of wing expansion. Neither of these defects are seen with two copies of P{UAS-EKO+}.
ShEKO.UAS.GFP(GL)/Scer\GAL4elav.PLu is a non-enhancer of short lived | dominant phenotype of AtpαDTS1
Scer\GAL4hs.2sev/ShEKO.UAS.GFP(GL) is a non-enhancer of short lived | dominant phenotype of AtpαDTS1
Scer\GAL4futsch-C380/ShEKO.UAS.GFP(GL) is a suppressor of abnormal neuroanatomy | larval stage phenotype of SERCAKum170, comt6
Scer\GAL4futsch-C380/ShEKO.UAS.GFP(GL) is a suppressor of abnormal neurophysiology | larval stage phenotype of SERCAKum170, comt6
ShEKO.UAS.GFP(GL), Scer\GAL4Tab2-201Y is a non-suppressor of abnormal neuroanatomy phenotype of Hsap\APPAβ42.UAS.Tag:SS(rPENK), Scer\GAL4Tab2-201Y
Scer\GAL4Ilp3.PB/ShEKO.UAS.GFP(GL) is a non-suppressor of decreased body size | adult stage phenotype of upd2Δ3-62
Scer\GAL4shot-OK307, ShEKO.UAS.GFP(GL) has wing phenotype, enhanceable by parats1
Scer\GAL4futsch-C380/ShEKO.UAS.GFP(GL) is a suppressor of neuromuscular junction | larval stage phenotype of SERCAKum170, comt6
ShEKO.UAS.GFP(GL), Scer\GAL4Tab2-201Y is a non-suppressor of Kenyon cell phenotype of Hsap\APPAβ42.UAS.Tag:SS(rPENK), Scer\GAL4Tab2-201Y
Scer\GAL4Ilp3.PB/ShEKO.UAS.GFP(GL) is a non-suppressor of wing phenotype of upd2Δ3-62
Expression of ShEKO.Scer\UAS.T:Avic\GFP-GL under the control of Scer\GAL4futsch-C380 suppresses the increased synaptic growth and transmitter release observed at the neuromuscular junction in comt6 Ca-P60AKum170 larvae.
There is no significant change in the lifespan of AtpαDTS1 mutants when they express ShEKO.Scer\UAS.T:Avic\GFP-GL under the control of either Scer\GAL4elav.PLu or Scer\GAL4hs.2sev.
79% of parats1 flies expressing two copies of P{UAS-EKO+} (driven by Scer\GAL4A307) fail to expand their wings.
Expression of ShEKO.Scer\UAS.T:Avic\GFP-GL does not suppress the learning defects seen in larvae expressing Hsap\APPAβ1-42.Scer\UAS.cIa pan-neuronally under the control of Scer\GAL4elav.PU. The progressive climbing defects seen in adult flies are also not suppressed.
Expression of ShEKO.Scer\UAS.T:Avic\GFP-GL does not suppress the mushroom body neuron loss seen in flies expressing Hsap\APPAβ1-42.Scer\UAS.cIa specifically in the mushroom body under the control of Scer\GAL4Tab2-201Y.
Expression of ShEKO.Scer\UAS.T:Avic\GFP-GL does not suppress the reduction in lifespan seen when Hsap\APPAβ1-42.Scer\UAS.cIa is expressed under the control of Scer\GAL4elav.PU.
Expression of ShEKO.Scer\UAS.T:Avic\GFP-GL under the control of Scer\GAL4Ilp3.PB does exacerbate the reduced wing area phenotype of upd2Δ3-62 mutants.
