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Cited by 276Sleep and Wakefulness ResearchCircadian rhythm and melatoninNeuroscience of respiration and sleep

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Galanin neurons in the ventrolateral preoptic area promote sleep and heat loss in mice

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Kroeger, Daniel, Absi, Gianna, Gagliardi, Celia, Bandaru, Sathyajit S., Madara, Joseph C., Ferrari, Loris L., Arrigoni, Elda, Münzberg, Heike, Scammell, Thomas E., Saper, Clifford B., Vetrivelan, Ramalingam·2018·Nature Communications
DOI 10.1038/s41467-018-06590-7PMC PMC6175893
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Abstract

The preoptic area (POA) is necessary for sleep, but the fundamental POA circuits have remained elusive. Previous studies showed that galanin (GAL)- and GABA-producing neurons in the ventrolateral preoptic nucleus (VLPO) express cFos after periods of increased sleep and innervate key wake-promoting regions. Although lesions in this region can produce insomnia, high frequency photostimulation of the POA GAL neurons was shown to paradoxically cause waking, not sleep. Here we report that photostimulation of VLPO GAL neurons in mice promotes sleep with low frequency stimulation (1–4 Hz), but causes conduction block and waking at frequencies above 8 Hz. Further, optogenetic inhibition reduces sleep. Chemogenetic activation of VLPO GAL neurons confirms the increase in sleep, and also reduces body temperature. In addition, chemogenetic activation of VLPO GAL neurons induces short-latency sleep in an animal model of insomnia. Collectively, these findings establish a causal role of VLPO GAL neurons in both sleep induction and heat loss.

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Fig. 1 Expression of AAV-ChR2 in VLPOGAL neurons in GAL-IRES-Cre mice. a Schematic representation of viral vector injections and implantation of optical fibers and EEG/EMG leads. b Outlines of viral injection sites in the VLPO at two levels (equivalent to AP: +0.26 and +AP: 0.14 in Ref. 70); Animal IDs (n = 10 mice) and the percentage increase in NREM sleep observed after photoactivation in each mouse is represented in matching colors. c cFos expression (black nuclei) in mCherry-expressing neurons (brown) after 2 h of optical stimulation (1 Hz, 10 ms) in VLPOGAL-mCherry mice (left) and VLPOGAL-ChR2 mice (right). 3 V, 3rd Ventricle; AC anterior commissure; OC optic chiasm; MnPO median preoptic nucleus; MPO medial preoptic area; PvPO periventricular preoptic area; VLPO ventrolateral preoptic area; VLPOc VLPO core; VLPOed VLPO extended dorsal; VLPOem VLPO extended medial. Scale bars in c are 200 μm (upper panels) and 50 μm (lower panels)
Fig. 1 Expression of AAV-ChR2 in VLPOGAL neurons in GAL-IRES-Cre mice. a Schematic representation of viral vector injections and implantation of optical fibers and EEG/EMG leads. b Outlines of viral injection sites in the VLPO at two levels (equivalent to AP: +0.26 and +AP: 0.14 in Ref. 70); Animal IDs (n = 10 mice) and the percentage increase in NREM sleep observed after photoactivation in each mouse is represented in matching colors. c cFos expression (black nuclei) in mCherry-expressing neurons (brown) after 2 h of optical stimulation (1 Hz, 10 ms) in VLPOGAL-mCherry mice (left) and VLPOGAL-ChR2 mice (right). 3 V, 3rd Ventricle; AC anterior commissure; OC optic chiasm; MnPO median preoptic nucleus; MPO medial preoptic area; PvPO periventricular preoptic area; VLPO ventrolateral preoptic area; VLPOc VLPO core; VLPOed VLPO extended dorsal; VLPOem VLPO extended medial. Scale bars in c are 200 μm (upper panels) and 50 μm (lower panels)CC BY · PubMed Central
Fig. 2 Optogenetic activation of VLPOGAL neurons increases NREM sleep. a In vitro stimulation of ChR2-expressing VLPOGAL neurons-one minute recordings with 10 ms pulses at 0.5, 1, 2, 4, 8, 16 Hz (middle column showing the first 30 s of stimulation); first three pulses (left column) and last three pulses (right column); dotted line represents the 0 mV level; laser stimulations in blue above traces. Note that stimulation rates of 8 Hz or above fail to elicit action potentials after the first laser pulses. b Quantification of stimulation-induced action potentials over the first minute of stimulation in a, one-way ANOVA for treatment, followed by Tukey’s post hoc test; F(5, 57) = 16.11, p < 0.0001 (n = 13 cells from three mice). c Frequency dependence of NREM sleep with optical stimulation (two-way repeated measures (RM) ANOVA for 2 h of recording time for ‘virus type’ and ‘stimulation frequency’ followed by Sidak’s post hoc test (virus type: F(1,14) = 31.69, p < 0.0001; stimulation frequency: F(6,84) = 5.809, p < 0.0001), n = 10 VLPOGAL-ChR2 mice vs. n = 6 VLPOGAL-mCherry mice). d Representative sample traces of EEG/EMG recordings during baseline (upper traces) and 1 Hz photostimulation of VLPOGAL neurons (lower traces); note large-amplitude and low frequency events in wake, NREM and REM sleep during stimulation; scale bar is 2 s. e EEG spectral analysis for NREM sleep, REM sleep and wake during 2 h of 1 Hz stimulation (Mann–Whitney test: NREM (Delta p = 0.0390, Theta p = 0.0390), REM (Delta p = 0.0028, Theta p = 0.1063), wake (Delta p = 0.0017, Theta p = 0.0110), n = 10 VLPOGAL-ChR2 mice vs. n = 6 VLPOGAL-mCherry mice). Data are Mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001
Fig. 2 Optogenetic activation of VLPOGAL neurons increases NREM sleep. a In vitro stimulation of ChR2-expressing VLPOGAL neurons-one minute recordings with 10 ms pulses at 0.5, 1, 2, 4, 8, 16 Hz (middle column showing the first 30 s of stimulation); first three pulses (left column) and last three pulses (right column); dotted line represents the 0 mV level; laser stimulations in blue above traces. Note that stimulation rates of 8 Hz or above fail to elicit action potentials after the first laser pulses. b Quantification of stimulation-induced action potentials over the first minute of stimulation in a, one-way ANOVA for treatment, followed by Tukey’s post hoc test; F(5, 57) = 16.11, p < 0.0001 (n = 13 cells from three mice). c Frequency dependence of NREM sleep with optical stimulation (two-way repeated measures (RM) ANOVA for 2 h of recording time for ‘virus type’ and ‘stimulation frequency’ followed by Sidak’s post hoc test (virus type: F(1,14) = 31.69, p < 0.0001; stimulation frequency: F(6,84) = 5.809, p < 0.0001), n = 10 VLPOGAL-ChR2 mice vs. n = 6 VLPOGAL-mCherry mice). d Representative sample traces of EEG/EMG recordings during baseline (upper traces) and 1 Hz photostimulation of VLPOGAL neurons (lower traces); note large-amplitude and low frequency events in wake, NREM and REM sleep during stimulation; scale bar is 2 s. e EEG spectral analysis for NREM sleep, REM sleep and wake during 2 h of 1 Hz stimulation (Mann–Whitney test: NREM (Delta p = 0.0390, Theta p = 0.0390), REM (Delta p = 0.0028, Theta p = 0.1063), wake (Delta p = 0.0017, Theta p = 0.0110), n = 10 VLPOGAL-ChR2 mice vs. n = 6 VLPOGAL-mCherry mice). Data are Mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001CC BY · PubMed Central
Fig. 3 Optogenetic inhibition of VLPOGAL neurons decreases NREM sleep. a Schematic representation of AAV injections and implantation of optical fibers and EEG/EMG leads. b In vitro whole-cell recordings in current clamp mode from VLPOGAL neurons expressing ArchT-GFP. Yellow/orange-light pulses (5 min duration) strongly hyperpolarized (difference in resting membrane potential between the last 10 s prior to laser pulse vs. the last 10 s of laser illumination was 28.4 ± 3.7 mV; p = 0.00026, paired t-test, n = 7 neurons in three mice) VLPOGAL neurons expressing ArchT and abolished action potentials (dotted line: 0 mV). c In vivo inhibition of VLPOGAL neurons by yellow/orange laser light (593.5 nm wavelength; ~10 mW/mm2 illumination at fiber tip) applied for 5 min every 30 min decreased the percent time spent in NREM sleep during the 12 h dark period. We compared 5 min periods ‘before’, ‘during’, and ‘after’ the photoinhibition using a one-way repeated measures ANOVA for ‘treatment groups’, followed by Tukey’s post hoc test (F(2, 12) = 15.86, p = 0.0005, n = 7 mice. d Sham photoinhibition did not alter the amount of NREM sleep in the same mice. Data are Mean ± SEM. ∗p < 0.05, ∗∗p < 0.01. Scale bars in b are 20 mV, 50 s
Fig. 3 Optogenetic inhibition of VLPOGAL neurons decreases NREM sleep. a Schematic representation of AAV injections and implantation of optical fibers and EEG/EMG leads. b In vitro whole-cell recordings in current clamp mode from VLPOGAL neurons expressing ArchT-GFP. Yellow/orange-light pulses (5 min duration) strongly hyperpolarized (difference in resting membrane potential between the last 10 s prior to laser pulse vs. the last 10 s of laser illumination was 28.4 ± 3.7 mV; p = 0.00026, paired t-test, n = 7 neurons in three mice) VLPOGAL neurons expressing ArchT and abolished action potentials (dotted line: 0 mV). c In vivo inhibition of VLPOGAL neurons by yellow/orange laser light (593.5 nm wavelength; ~10 mW/mm2 illumination at fiber tip) applied for 5 min every 30 min decreased the percent time spent in NREM sleep during the 12 h dark period. We compared 5 min periods ‘before’, ‘during’, and ‘after’ the photoinhibition using a one-way repeated measures ANOVA for ‘treatment groups’, followed by Tukey’s post hoc test (F(2, 12) = 15.86, p = 0.0005, n = 7 mice. d Sham photoinhibition did not alter the amount of NREM sleep in the same mice. Data are Mean ± SEM. ∗p < 0.05, ∗∗p < 0.01. Scale bars in b are 20 mV, 50 sCC BY · PubMed Central
Fig. 4 Chemoactivation of VLPOGAL neurons by CNO. a Schematic representation of viral vector injections. b Outlines of AAV-hM3Dq injection sites in VLPO at two levels (equivalent to AP: +0.26 and AP: +0.14 in Ref. 70; Animal IDs (n = 15 mice) and the magnitude of hypothermia observed after chemoactivation in each mouse is represented in matching colors c cFos expression (black nuclei) in mCherry-expressing neurons (brown) 3 h after i.p. injections of CNO (0.3 mg/kg) in VLPOGAL-mCherry mice (left) and VLPOGAL-hM3Dq mice (right). d Representative trace of in vitro activation of an hM3Dq-expressing VLPOGAL neuron with 1 μM CNO (n = 5 cells from five mice). 3 V, 3rd Ventricle; AC Anterior commissure; OC optic chiasm; MnPO Median preoptic nucleus; MPO Medial preoptic area; PvPO Periventricular preoptic area; VLPO Ventrolateral preoptic area; VLPOc. VLPO core; VLPOed, VLPO extended dorsal; VLPOem, VLPO extended medial. Scale bars in c 200 μm (upper panels) and 50 μm (lower panels) and d 20 mV, 60 s
Fig. 4 Chemoactivation of VLPOGAL neurons by CNO. a Schematic representation of viral vector injections. b Outlines of AAV-hM3Dq injection sites in VLPO at two levels (equivalent to AP: +0.26 and AP: +0.14 in Ref. 70; Animal IDs (n = 15 mice) and the magnitude of hypothermia observed after chemoactivation in each mouse is represented in matching colors c cFos expression (black nuclei) in mCherry-expressing neurons (brown) 3 h after i.p. injections of CNO (0.3 mg/kg) in VLPOGAL-mCherry mice (left) and VLPOGAL-hM3Dq mice (right). d Representative trace of in vitro activation of an hM3Dq-expressing VLPOGAL neuron with 1 μM CNO (n = 5 cells from five mice). 3 V, 3rd Ventricle; AC Anterior commissure; OC optic chiasm; MnPO Median preoptic nucleus; MPO Medial preoptic area; PvPO Periventricular preoptic area; VLPO Ventrolateral preoptic area; VLPOc. VLPO core; VLPOed, VLPO extended dorsal; VLPOem, VLPO extended medial. Scale bars in c 200 μm (upper panels) and 50 μm (lower panels) and d 20 mV, 60 sCC BY · PubMed Central
Fig. 5 Chemoactivation of VLPOGAL neurons promotes NREM sleep and hypothermia. a, b Percentages of NREM sleep every 4 h for 24 h after saline or CNO administration (red arrow) at 19:00 h a or 10:00 h b and respective NREM sleep latencies. Two-way repeated measures (RM) ANOVA for the first 12 h after treatment for ‘time’ and ‘compound injected’, followed by Sidak’s post hoc test (19:00 h injections (n = 9 mice): time F(2,32) = 2.398, p = 0.1070, compound injected F(1,16) = 22.76, p = 0.0002; 10:00 h injections (n = 13 mice): time F(2,48) = 77.29, p < 0.0001, compound injected F(1,24) = 28.92, p < 0.0001). c, d Percentages of REM sleep after saline or CNO at 19:00 h c or 10:00 h d with respective REM sleep latencies. Two-way RM ANOVA followed by Sidak’s post hoc test (19:00 h injections (n = 9 mice): time F(2,32) = 10.77, p = 0.0003, compound injected F(1,16) = 7.763, p = 0.0132; 10:00 h injections (n = 13 mice): time F(2,48) = 11.75, p < 0.0001, compound injected F(1,24) = 41.06, p < 0.0001); Wilcoxon matched-pairs signed-rank test for REM sleep latencies (19:00 h injections p = 0.0010; 10:00 h injections p = 0.0002). e, f Tb after saline or CNO at 19:00 h e or 10:00 h f; dotted lines represent SEM; Two-way RM ANOVA for the first 12 h after treatment for ‘time’ and ‘compound injected’, followed by Sidak’s post hoc test (19:00 h injections: time F(143, 4004) = 19.4, p < 0.0001, compound injected F(1,28) = 19.09, p = 0.0002; n = 15 mice; 10:00 h injections: time F(143,4290) = 65, p < 0.0001, compound injected F(1,30) = 29.78, p < 0.0001; n = 16 mice). g, i Correlation between percentage change in NREM sleep and fall in Tb after CNO at 19:00 h g or 10:00 h i; Pearson correlation (19:00 h injections: r = 0.5831, p = 0.1292, n = 8 mice; 10:00 h injections: r = 0.7076, p = 0.01; n = 12 mice). h, j Correlation between percentage change in REM sleep and fall in Tb after CNO at 19:00 h h or 10:00 j; Pearson correlation (19:00 h injections r = 0.4093, p = 0.3140, n = 8 mice; 10:00 injections: r = 0.5749, p = 0.0505; n = 12 mice). Red arrows indicate saline/CNO injections. Data are Mean ± SEM, except sleep latencies which are Mean ± SD, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001
Fig. 5 Chemoactivation of VLPOGAL neurons promotes NREM sleep and hypothermia. a, b Percentages of NREM sleep every 4 h for 24 h after saline or CNO administration (red arrow) at 19:00 h a or 10:00 h b and respective NREM sleep latencies. Two-way repeated measures (RM) ANOVA for the first 12 h after treatment for ‘time’ and ‘compound injected’, followed by Sidak’s post hoc test (19:00 h injections (n = 9 mice): time F(2,32) = 2.398, p = 0.1070, compound injected F(1,16) = 22.76, p = 0.0002; 10:00 h injections (n = 13 mice): time F(2,48) = 77.29, p < 0.0001, compound injected F(1,24) = 28.92, p < 0.0001). c, d Percentages of REM sleep after saline or CNO at 19:00 h c or 10:00 h d with respective REM sleep latencies. Two-way RM ANOVA followed by Sidak’s post hoc test (19:00 h injections (n = 9 mice): time F(2,32) = 10.77, p = 0.0003, compound injected F(1,16) = 7.763, p = 0.0132; 10:00 h injections (n = 13 mice): time F(2,48) = 11.75, p < 0.0001, compound injected F(1,24) = 41.06, p < 0.0001); Wilcoxon matched-pairs signed-rank test for REM sleep latencies (19:00 h injections p = 0.0010; 10:00 h injections p = 0.0002). e, f Tb after saline or CNO at 19:00 h e or 10:00 h f; dotted lines represent SEM; Two-way RM ANOVA for the first 12 h after treatment for ‘time’ and ‘compound injected’, followed by Sidak’s post hoc test (19:00 h injections: time F(143, 4004) = 19.4, p < 0.0001, compound injected F(1,28) = 19.09, p = 0.0002; n = 15 mice; 10:00 h injections: time F(143,4290) = 65, p < 0.0001, compound injected F(1,30) = 29.78, p < 0.0001; n = 16 mice). g, i Correlation between percentage change in NREM sleep and fall in Tb after CNO at 19:00 h g or 10:00 h i; Pearson correlation (19:00 h injections: r = 0.5831, p = 0.1292, n = 8 mice; 10:00 h injections: r = 0.7076, p = 0.01; n = 12 mice). h, j Correlation between percentage change in REM sleep and fall in Tb after CNO at 19:00 h h or 10:00 j; Pearson correlation (19:00 h injections r = 0.4093, p = 0.3140, n = 8 mice; 10:00 injections: r = 0.5749, p = 0.0505; n = 12 mice). Red arrows indicate saline/CNO injections. Data are Mean ± SEM, except sleep latencies which are Mean ± SD, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001CC BY · PubMed Central
Fig. 6 Warm ambient temperatures (Ta) enhance NREM sleep with chemoactivation of VLPOGAL neurons. a Schematic of experimental paradigm: baseline Ta 22 °C and exposure to Ta 29 °C and 36 °C. b Mean Tb for 2 h after saline/CNO (0.3 mg/kg) injections at 19:00 h in different Ta: Two-way repeated measures (RM) ANOVA for 2 h after treatment for ‘Ta’ and ‘compound injected’, followed by Sidak’s post hoc test (Ta: F(2,28) = 46.87, p < 0.0001, compound injected: F(1,14) = 26.26, p = 0.0002; n = 8 mice). c Percentages of NREM sleep for 2 h after saline or CNO treatment in different Ta: Two-way RM ANOVA followed by Sidak’s post hoc test (Ta: F(2,28) = 28.29, p < 0.0001, compound injected: F(1,14) = 40.08, p < 0.0001; n = 8 mice). d NREM latencies after saline or CNO injections in different Ta: Two-way RM ANOVA followed by Sidak’s post hoc test (Ta: F(2,28) = 0.1369, p = 0.0002, compound injected: F(1,14) = 5.12, p = 0.0401; n = 8 mice). e CNO/Saline ratio of NREM delta power for 2 h after saline or CNO treatment in different Ta. One-way ANOVA (Friedman test) for 2 h after treatment for ‘Ta‘, followed by Dunn’s multiple comparisons test (p = 0.0303); f Percentages of REM sleep for 2 h after saline or CNO treatment in different Ta: Two-way RM ANOVA followed by Sidak’s post hoc test (Ta: F(2,28) = 3.228, p = 0.0548, compound injected: F(1,14) = 20.19, p = 0.0005; n = 8 mice). g Cumulative REM sleep amounts every 4 h for 24 h after saline or CNO at Ta 29 °C during the light period (10:00 h). Note that animals at 29 °C who received CNO had less REM sleep in the initial 4 hours, but by the end of the next 16 hours REM rebound resulted in catching up to animals treated with saline: Two-way RM ANOVA for ‘time’ and ‘compound injected’, followed by Sidak’s post hoc test (time: F(5,170) = 391.5, p < 0.0001, compound injected: F(3,34) = 12.79, p < 0.0001; n = 13 mice (Ta 22 °C), n = 6 mice (Ta 29 °C). Data are Mean ± SEM, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001
Fig. 6 Warm ambient temperatures (Ta) enhance NREM sleep with chemoactivation of VLPOGAL neurons. a Schematic of experimental paradigm: baseline Ta 22 °C and exposure to Ta 29 °C and 36 °C. b Mean Tb for 2 h after saline/CNO (0.3 mg/kg) injections at 19:00 h in different Ta: Two-way repeated measures (RM) ANOVA for 2 h after treatment for ‘Ta’ and ‘compound injected’, followed by Sidak’s post hoc test (Ta: F(2,28) = 46.87, p < 0.0001, compound injected: F(1,14) = 26.26, p = 0.0002; n = 8 mice). c Percentages of NREM sleep for 2 h after saline or CNO treatment in different Ta: Two-way RM ANOVA followed by Sidak’s post hoc test (Ta: F(2,28) = 28.29, p < 0.0001, compound injected: F(1,14) = 40.08, p < 0.0001; n = 8 mice). d NREM latencies after saline or CNO injections in different Ta: Two-way RM ANOVA followed by Sidak’s post hoc test (Ta: F(2,28) = 0.1369, p = 0.0002, compound injected: F(1,14) = 5.12, p = 0.0401; n = 8 mice). e CNO/Saline ratio of NREM delta power for 2 h after saline or CNO treatment in different Ta. One-way ANOVA (Friedman test) for 2 h after treatment for ‘Ta‘, followed by Dunn’s multiple comparisons test (p = 0.0303); f Percentages of REM sleep for 2 h after saline or CNO treatment in different Ta: Two-way RM ANOVA followed by Sidak’s post hoc test (Ta: F(2,28) = 3.228, p = 0.0548, compound injected: F(1,14) = 20.19, p = 0.0005; n = 8 mice). g Cumulative REM sleep amounts every 4 h for 24 h after saline or CNO at Ta 29 °C during the light period (10:00 h). Note that animals at 29 °C who received CNO had less REM sleep in the initial 4 hours, but by the end of the next 16 hours REM rebound resulted in catching up to animals treated with saline: Two-way RM ANOVA for ‘time’ and ‘compound injected’, followed by Sidak’s post hoc test (time: F(5,170) = 391.5, p < 0.0001, compound injected: F(3,34) = 12.79, p < 0.0001; n = 13 mice (Ta 22 °C), n = 6 mice (Ta 29 °C). Data are Mean ± SEM, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001CC BY · PubMed Central
Fig. 7 Chemogenetic activation of VLPOGAL neurons attenuates sleep-onset insomnia. a Schematic of experimental setup showing mice introduced to a new cage with fresh nesting material immediately after i.p. injections of either saline or CNO. b Percentages of NREM sleep every 2 h for 12 h after saline or CNO (0.3 mg/kg) injections and cage change at 10:00 h. Typically, mice introduced to a new cage during the day (when they usually sleep about 60% of the time) have reduced sleep for 2–4 h with a prolonged latency to first NREM sleep. This is rescued by activation of VLPOGAL neurons. Two-way repeated measures (RM) ANOVA for the first 6 h after treatment for ‘time’ and ‘compound injected’, followed by Sidak’s post hoc test (time: F(3,36) = 64.72, p < 0.0001, compound injected: F(1,12) = 8.644, p = 0.0124; n = 7 mice). c NREM sleep latencies after injections and cage change: Wilcoxon matched-pairs signed-rank test (p = 0.0156; n = 7 mice). d Tb after injections and cage change. Two-way RM ANOVA for the first 6 h after treatment for ‘time’ and ‘compound injected’, followed by Sidak’s post hoc test (time: F(71,852) = 14.72, p < 0.0001, compound injected: F(1,12) = 15.21, p = 0.0021; n = 7 mice). Red arrows indicate saline/CNO injections. The data are Mean ± SEM, except sleep latencies which are Mean ± SD, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001
Fig. 7 Chemogenetic activation of VLPOGAL neurons attenuates sleep-onset insomnia. a Schematic of experimental setup showing mice introduced to a new cage with fresh nesting material immediately after i.p. injections of either saline or CNO. b Percentages of NREM sleep every 2 h for 12 h after saline or CNO (0.3 mg/kg) injections and cage change at 10:00 h. Typically, mice introduced to a new cage during the day (when they usually sleep about 60% of the time) have reduced sleep for 2–4 h with a prolonged latency to first NREM sleep. This is rescued by activation of VLPOGAL neurons. Two-way repeated measures (RM) ANOVA for the first 6 h after treatment for ‘time’ and ‘compound injected’, followed by Sidak’s post hoc test (time: F(3,36) = 64.72, p < 0.0001, compound injected: F(1,12) = 8.644, p = 0.0124; n = 7 mice). c NREM sleep latencies after injections and cage change: Wilcoxon matched-pairs signed-rank test (p = 0.0156; n = 7 mice). d Tb after injections and cage change. Two-way RM ANOVA for the first 6 h after treatment for ‘time’ and ‘compound injected’, followed by Sidak’s post hoc test (time: F(71,852) = 14.72, p < 0.0001, compound injected: F(1,12) = 15.21, p = 0.0021; n = 7 mice). Red arrows indicate saline/CNO injections. The data are Mean ± SEM, except sleep latencies which are Mean ± SD, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001CC BY · PubMed Central

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Authors
Kroeger, D., Absi, G., Gagliardi, C., Bandaru, S. S., Madara, J. C., Ferrari, L. L., Arrigoni, E., Münzberg, H., Scammell, T. E., Saper, C. B., & Vetrivelan, R.
Affiliations
  • Beth Israel Deaconess Medical Center (US)
  • Pennington Biomedical Research Center (US)
Corresponding author
  • Saper, Clifford B.
  • Vetrivelan, Ramalingam
ORCID
Venue
Nature Communications
Publisher
Springer Science and Business Media LLC
Published
2018-10-08
Volume
9
Issue
1
Pages
4129-4129
ISSN
2041-1723 (ISSN-L)
DOI
10.1038/s41467-018-06590-7
PMC
PMC6175893
OpenAlex
W2895180682
Cited by
276
References
70
Citations by year
2026: 26 · 2025: 29 · 2024: 46 · 2023: 37 · 2022: 32 · 2021: 46 · 2020: 29 · 2019: 31
FWCI
15.78
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature Communications (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • RePEc: Research Papers in Economics (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
Office of Extramural Research, National Institutes of Health (NS091126, NS085477, HL095491, AG009975)
Topics
  • Life Sciences › Neuroscience › Cognitive Neuroscience › Sleep and Wakefulness Research
  • Life Sciences › Neuroscience › Endocrine and Autonomic Systems › Circadian rhythm and melatonin
  • Life Sciences › Neuroscience › Endocrine and Autonomic Systems › Neuroscience of respiration and sleep
Keywords
ventrolateral preoptic area, sleep regulation, heat loss, body temperature, photostimulation, optogenetics, chemogenetics, insomnia, c-fos, wakefulness, neuronal firing rate
MeSH terms
Animals, Body Temperature Regulation, Electroencephalography, Electromyography, Sleep Initiation and Maintenance Disorders, Male, Mice, Transgenic, Neurons, Preoptic Area, Sleep, Galanin, Gene Expression Profiling

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