A Toll-receptor map underlies structural brain plasticity
- Neurodevelopment Lab, School of Biosciences, University of Birmingham, United Kingdom
- Rudolf Virchow Center for Experimental Biomedicine, University of Würzburg, Germany
- Facultad de Ingeniería, Universidad de Ibagué, Colombia
- Neuroscience, University of Arizona College of Science, United States
Figures
Figure 1 with 1 supplement
Expression of Tolls demarcates the anatomical map of the adult brain.
(A) Toll receptor expression visualised with: Anti-Toll-1 antibodies, in retinal photoreceptors; CRISPR/Cas9 generated Toll-2pTVGAL4 > FlyBow, throughout the brain; MIMIC-RMCE generated Toll-3GAL4 > …
Figure 1—figure supplement 1
Most Tolls are expressed in the central nervous system.
Reverse transcription PCR (RT-PCR) carried out to detect transcripts for Tolls in whole embryos, and dissected central nervous systems (CNS) of larvae and pupae, and dissected adult brains. Control …
Figure 2 with 1 supplement
Toll-2 knock-down caused neuronal loss in the pupal and adult brain.
(A,B) Overlapping but distinct expression of Toll-2 and the adaptor MyD88, visualised with MyD88 >tdTomato, Toll-2pTVGAL4 > UASFlyBow, MyD88 >histone YFP and Toll-2pTVGAL4 > UAShistone YFP. Toll-2 …
Figure 2—figure supplement 1
Over-expression of Toll-2 does not affect Kenyon cell number.
(A) The MBGAL4 line drives expression virtually only in Kenyon cells (MB-GAL4 >UASFlyBow). (B) The ROI indicates MBGAL4 >UAS HisYFP cells counted automatically with DeadEasy Kenyon cells software in …
Figure 3 with 1 supplement
Loss of Toll-2 function caused neurodegeneration and impaired behaviour.
(A) Toll-2 is expressed in pupal brains, most prominently in optic lobes, mushroom bodies and SOG. (B–H) Toll-2pTV mutant MARCM clones - labelled with GFP - induced in pupa cause neuronal loss …
Figure 3—figure supplement 1
Toll-2 loss of function compromises survival, longevity and locomotion.
(A) Complementation tests show Toll-2pTV is a loss of function allele, as it causes semi-lethality over the known null Toll-2 allele 18wD7-35 and over deficiency Df(2R)BSC594 for the locus, but not …
Figure 4 with 1 supplement
Cell number is regulated by Toll-2 and neuronal activity at the adult critical period.
At adult days 0–2 post-eclosion, within the critical period: (A) Conditional over-expression of Toll-2 increased MyD88 >hisYFP+ cell number in the central brain. Cells were counted automatically in …
Figure 4—figure supplement 1
Toll-signalling downstream factors were found in the brain.
(A,B) sarm was expressed in the adult brain: line sarmGAL4NP760 drives expression of four short isoforms, and sarmGAL4NP0257 of the longest isoform, visualised with UAS-FlyBow. Note prominent …
Figure 5
Toll-2 can induce cell proliferation in the brain.
(A) Toll-2 induces G1/S cycling. Over-expression of the G1/S inhibitor Rbf280 in Toll-2+ cells reduced Toll-2 >his-YFP+ KC cell number by day one pupa, compared to controls, meaning that Toll-2+ is …
Figure 6
MyD88 and Toll-2 are expressed in progenitor cells in the adult brain.
(A–D, K) FUCCI expressed with tubGAL80ts, MyD88GAL4 revealed cycling cells in the adult brain. Arrows point to GFP— RFP+ cells in S-phase, GFP+RFP— cells in G1, and GFP+RFP+ cells in G2/M. (D) …
Figure 7
Toll-2 regulates Yki in the adult brain.
(A–D) Nuclear translocation of a Yki-GFP fusion protein at the adult brain critical period: (A) there were many cells with nuclear Yki-GFP in the adult brain, particularly upon conditional Tolll-2 …
Figure 8 with 1 supplement
Tolls regulate brain size in the pupal and adult brain.
(A,B) Constant RNAi knock-down of three Tolls with Toll-2GAL4 resulted in smaller brains. Optic lobes were smaller than in controls, and so were central brains (Figure 8—figure supplement 1A). (B) …
Figure 8—figure supplement 1
Conditional knock-down of Toll-2 and −6 in the adult brain altered cell number and brain size.
(A) Triple Toll RNAi knock-down with Toll-2GAL4 reduced central brain area, meaning it affects the central brain as well as the optic lobes shown in Figure 8A,B (images shown in Figure 8A). …
Figure 9
Toll receptors topographically modulate experience-dependent structural brain plasticity.
(A) Experience – that is stimulation through vision - alters the size of multiple brain domains, including optic lobes (for vision), central brain (integration, equivalent to mammalian cortex) and …
Additional files
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Supplementary file 1
Key Resources Table.
- https://cdn.elifesciences.org/articles/52743/elife-52743-supp1-v2.docx
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Supplementary file 2
Genotypes, samples size and statistical analysis.
All details of the quantitative analyses including: genotypes, sample sizes, statistical tests carried out: normality tests, parametric and non-parametric tests for two or more samples, test results and p-value for whole data sets, and multiple comparison post-hoc correction tests and their p-values.
- https://cdn.elifesciences.org/articles/52743/elife-52743-supp2-v2.xlsx
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Supplementary file 3
List of primers.
Primers used for all cloning and RT-PCR experiments.
- https://cdn.elifesciences.org/articles/52743/elife-52743-supp3-v2.docx
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Transparent reporting form
- https://cdn.elifesciences.org/articles/52743/elife-52743-transrepform-v2.docx
