Hypocretin neuron-specific transcriptome profiling identifies the sleep modulator Kcnh4a
- Bar-Ilan University, Israel
- Toldot Genetics Ltd., Israel
- The Weizmann Institute of Science, Israel
- Massachusetts Institute of Technology, United states
Figures
Figure 1
Isolation of Hcrt neurons, RNA-seq, and experimental design.
(A) Dorsal view of 6 dpf hcrt:EGFP larvae. (B) Dorsal view of the hypothalamus region of 6 dpf hcrt:EGFP larvae expressing EGFP in Hcrt neurons. (C) Cell suspension from the whole head of 6 dpf hcrt:…
Figure 2
The expression pattern of selected candidate Hcrt-neuron–specific genes.
(A) Table presenting the top 20 Hcrt-enriched transcripts. (B–M) Dorsal view of whole-mount ISH-stained 2 dpf WT larvae. Based on the RNA-seq and the bioinformatic analysis, the expression pattern …
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Figure 2—source data 1
Hcrt-neuron enriched transcripts.
All the genes detected have p <0.01 with Bonferroni correction for multiple testing (Materials and methods).
- https://doi.org/10.7554/eLife.08638.005
Figure 3
Selected candidate genes are expressed in Hcrt neurons.
(A–I'') Double fluorescent staining of the candidate genes (red) and EGFP (green) was performed in 2 dpf hcrt:EGFP larvae using whole-mount ISH and immunofluorescence, respectively. White arrows …
Figure 4
Candidate genes are expressed in cell populations located adjacent to Hcrt neurons.
(A–I'') Fluorescent ISH and immunofluorescence experiments in 2 dpf larvae and adult hcrt:EGFP zebrafish showing three candidate genes that are expressed adjacently to Hcrt neurons within the …
Figure 5
Predicted TFs that regulate the expression of Hcrt-neuron–specific transcripts.
(A) TFs with a p <0.005 and their target Hcrt-neuron–specific genes. For each of these transcription factors, a combined score for each gene is calculated according to all of the predicted binding …
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Figure 5—source data 1
Predicted Hcrt-neuron enriched transcription factors and their target genes.
- https://doi.org/10.7554/eLife.08638.009
Figure 6 with 1 supplement
The genomic location, phylogenetic reconstruction and structure of kcnh4a, and the generation of kcnh4a-/- zebrafish.
(A) Synteny analysis shows similar genomic context of hcrt in zebrafish and mammals. Notably, kcnh4a is located a few kbs downstream to hcrt in zebrafish and mammals. (B) The 16-exon kcnh4a gene …
Figure 6—figure supplement 1
Hypothalamic kcnh4a-expressing neurons are glutamatergic.
Double in-situ hybridization against kcnh4a (red, A, B) and gad67 (green, A'), or vglut2b (green, B'). All pictures are on a single optical plane of 0.5 µm.
Figure 7 with 1 supplement
Sleep time and quality are reduced in kcnh4a-/- larvae during the night.
(A) The locomotor activity of kcnh4a-/- (n=85), kcnh4a+/- (n=208), and kcnh4a+/+ (n=98) is shown. kcnh4a-/- larvae exhibit increased locomotor activity compared with kcnh4a+/- and kcnh4a / under LD …
Figure 7—figure supplement 1
Sleep time is increased following sleep deprivation (SD).
(A) At 6dpf, larvae were sleep deprived for 6 hr during the night under constant dark conditions (DD) and sleep time was monitored in the following nine hours. (B,C) Sleep was recovered in …
