Article Figures & Data
Figures
- Figure 1.
Sleep–wake states and GABA receptors in Orx and MCH neurons across groups. A, Percentage of time spent in wake during the 2 h preceding termination in SC, SD, and SR groups. Percentage wake is significantly higher in SD compared to SC and SR and significantly lower in SR compared to SC. B, Proportion of Orx+ or MCH+ neurons bearing GABAARs across groups. The percentage Orx+/GABAAR+ was significantly greater in SD compared to SC and SR (B1), whereas the percentage MCH+/GABAAR+ neurons was insignificantly less in SD compared to SC and SR (B2). C, Luminance of the GABAAR immunofluorescence on Orx and MCH neurons across groups, which was significantly increased on the Orx+ neurons (C1) and decreased on the MCH+ neurons (C2) in SD compared to SC and SR. D, Proportion of the Orx (D1) and MCH (D2) neurons expressing GABABRs, which did not change across groups. E, Luminance of the GABABR which was significantly higher in Orx+ neurons (E1), and significantly lower in MCH+ neurons (E2) following SD compared to SC and SR. Note that the changes in GABARs on Orx neurons parallel the percentage wake, whereas those on MCH+ neurons parallel the percentage sleep across groups. *Indicates significant difference of SD relative to SC and SR (p < 0.05). §Indicates significant difference of SR relative to SC (p < 0.05), according to post hoc paired comparisons following one-way ANOVA (Table 1). Abbreviations: Lum, luminance.
- Figure 2.
GABAARs in Orx neurons across groups. Confocal images of immunostained sections indicate that the GABAAR (red) was minimal on Orx+ neurons (blue, indicated by filled arrowheads) compared to that on adjacent Orx-negative neurons (stained with FNS, green, indicated by carets). A, The GABAAR immunofluorescence was minimally visible as small clusters along a portion of the plasma membrane of an Orx+ cell body in an SC mouse, in which it was readily visible along the full membrane of an Orx-negative cell body. B, The GABAAR staining was more visible as larger clusters along a larger portion of the membrane of an Orx+ cell in an SD mouse. C, The GABAAR staining was similar in an SR mouse to that in SC. Scale bars, 10μm. Image thickness: A, B, 1500 nm; C, 2000 nm.
- Figure 3.
GABAARs in MCH neurons across groups. Confocal images indicate that GABAAR immunostaining (red) was moderate in MCH neurons (blue, indicated by filled arrowheads) though less than that in some adjacent MCH-negative neurons (stained with FNS, green, indicated by carets). A, GABAAR immunostaining was present as clusters visible along the full plasma membrane of the cell body in an SC mouse. B, GABAAR immunostaining was barely visible on MCH+ neurons, whereas it was prominent on adjacent MCH-negative neuron in an SD mouse. C, GABAAR immunostaining appeared to be moderate in an SR mouse, similar to that in SC. Scale bars, 10μm. Image thickness, 500 nm.
- Figure 4.
GABABRs in Orx neurons across groups. Confocal images of the GABABR immunostaining (red) in Orx neurons (blue, indicated by filled arrowheads). A, The GABABR immunofluorescence was minimally visible as clusters over the cytoplasm of an Orx+ cell body in an SC mouse. B, The GABABR staining was more visible as larger clusters over the cytoplasm and partially on the plasma membrane of an Orx+ cell in an SD mouse. C, The GABABR staining was similar in an SR mouse to that in SC. Scale bars, 10μm. Image thickness, 1500 nm.
- Figure 5.
GABABRs in MCH neurons across groups. Confocal images of the GABABR immunostaining (red) in MCH neurons (blue, indicated by filled arrowheads). A, GABABR immunostaining was present as prominent large clusters over the cytoplasm of an MCH+ neuron in an SC mouse. B, The GABABR immunostaining was minimally visible over the cytoplasm of an MCH+ neuron in an SD mouse. C, The GABABR immunostaining appeared to be prominent over the cytoplasm and near the plasma membrane of an SR mouse, similar to that in SC. Scale bars, 10μm. Image thickness: A, 2000 nm; B, C, 1500 nm.
Tables
Dataset Figure One-way ANOVA (group = 3 levels) F value df; group, error p value Tukey’s HSD paired-comparisons p value SC-SD SC-SR SD-SR % Wake 1A 1032.33 2, 22 <0.001 <0.001* <0.001§ <0.001* % GABAAR+/Orx+ 1B1 10.27 2, 9 0.005 0.009* 0.910 0.010* % GABAAR+/MCH+ 1B2 1.48 2, 9 0.270 n/a n/a n/a Lum GABAAR:Orx+ 1C1 7.43 2, 115 0.001 0.010* 0.800 0.002* Lum GABAAR:MCH+ 1C2 5.88 2, 117 0.004 0.010* 0.970 0.020* % GABABR+/Orx+ 1D1 n/a n/a n/a n/a n/a n/a % GABABR+/MCH+ 1D2 n/a n/a n/a n/a n/a n/a Lum GABABR:Orx+ 1E1 15.93 2, 127 <0.001 <0.001* 0.970 <0.001* Lum GABABR:MCH+ 1E2 27.16 2, 127 <0.001 <0.001* 0.860 <0.001*
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