Distinct ipRGC subpopulations mediate light’s acute and circadian effects on body temperature and sleep
- Johns Hopkins University, United States
- Northwestern University, United States
Figures
Figure 1 with 2 supplements
Melanopsin mediates the acute effects of light on body temperature.
(A) Paradigm to measure body temperature continuously in a 12:12 light dark cycle with a 3 hr light pulse at ZT14. (B) 48 hr of continuous body temperature monitoring in wildtype male mice (n = 13) …
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Figure 1—source data 1
Temperature data for Figure 1.
- https://doi.org/10.7554/eLife.44358.006
Figure 1—figure supplement 1
Intensity-dependent decrease in core body temperature during a nocturnal light pulse.
(A) Experimental paradigm consisting of a 12 hr/12 hr light/dark cycle with a single 3 hr light pulse starting at Zeitgeber time (ZT) 14 (i.e. 2 hr after lights-off). Each experimental night, a …
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Figure 1—figure supplement 1—source data 1
Temperature data for Figure 1—figure supplement 1.
- https://doi.org/10.7554/eLife.44358.007
Figure 1—figure supplement 2
Melanopsin-dependence of light-induced body temperature changes.
Relative change from ZT14 in mean body temperature in Wildtype (n = 15), Melanopsin-only (Gnat1-/-;Gnat2-/-, n = 11) or Melanopsin KO (Opn4-/-, n = 6) for the 3 hr during the light pulse (Light) or …
Figure 2 with 1 supplement
Brn3b-negative ipRGCs are insufficient for acute body temperature regulation via the SCN.
(A) Diurnal body temperature in control (Opn4Cre/+, n = 9) and (B) Brn3b-DTA (Opn4Cre/+;Brn3bDTA/+, n = 7). (C) Diurnal body temperature amplitude in the two groups. p=0.223 by t-test. (D) Body …
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Figure 2—source data 1
Temperature data for Figure 2.
- https://doi.org/10.7554/eLife.44358.010
Figure 2—figure supplement 1
Brn3b-DTA body temperature regulation with light.
Relative change from ZT14 in mean body temperature in Control (Opn4Cre/+, n = 9) or Brn3b-DTA mice (Opn4Cre/+;Brn3bzDTA/+, n = 7) for the 3 hr during the light pulse (Light) or during the previous …
Figure 3 with 2 supplements
Activation of Brn3b-positive RGCs is sufficient to drive sustained body temperature decreases.
(A) Diagram of intravitreal delivery of AAV2-hSyn-DIO-hM3Dq-mCherry to Brn3bCre/+ mice, and confirmation of infection of ipRGCs. (B) 54 hr continuous diurnal body temperature recordings in …
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Figure 3—source data 1
Temperature data for Figure 3.
- https://doi.org/10.7554/eLife.44358.014
Figure 3—figure supplement 1
Brn3b-Cre::hM3D(Gq) expression and control experiments.
(A) mCherry expression (magenta) from Brn3b-Cre with AAV-DIO-hM3D(Gq)-mCherry injected in to the retina and colocalized with melanopsin (OPN4, green). Scale bar = 50 µm. (B) Quantification of …
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Figure 3—figure supplement 1—source data 1
Temperature data for Figure 3—figure supplement 1.
- https://doi.org/10.7554/eLife.44358.015
Figure 3—figure supplement 2
No effect of CNO on body temperature in wildtype mice.
(A) Body temperature of wildtype mice (n = 9) was monitored continuously and PBS was injected on night one at ZT14, followed by CNO injection (1 mg/kg) on night two at ZT14. (B,C) Normalized body …
Figure 4 with 3 supplements
Brn3b-positive ipRGCs are not required for circadian photoentrainment of sleep, but are required for its acute induction by light.
(A–C) Percent time spent asleep in 1 hr bins across the 24 hr day for (A) Control (black) mice (n = 14) and (B) Brn3b-DTA (blue) mice (n = 13) lacking Brn3b-positive ipRGCs. Both lines showed normal …
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Figure 4—source data 1
Sleep data for Figure 4.
- https://doi.org/10.7554/eLife.44358.020
Figure 4—figure supplement 1
NREM and REM measurements in Control and Brn3b-DTA mice.
(A) Percent sleep recorded across the 24 hour day as NREM vs. REM in Control (black) and Brn3b-DTA mice (blue). Brn3b-DTA mice showed a small but significant increase in NREM and decrease in REM …
Figure 4—figure supplement 2
Brn3b-DTA body temperature regulation with light.
Relative change in sleep time in Control (Opn4Cre/+, n = 14) or Brn3b-DTA mice (Opn4Cre/+;Brn3bzDTA/+, n = 13) for the 3 hr during the light pulse or during the previous control night (ZT14-17). …
Figure 4—figure supplement 3
Wheel-running activity in Brn3b-DTA mice.
(A) Control (Opn4Cre/+, n = 5) and (B) Brn3b-DTA (Opn4Cre/+;Brn3bDTA/+, n = 6) were housed in a 12:12 LD cycle and subjected to a 3 hr light pulse starting at ZT14. (C) Activity counts in 30 min …
Author response image 1
Schematic of analyses throughout the manuscript with example graphs from Brn3bDTA analyses.
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers |
|---|---|---|---|
| Genetic reagent (Mus musculus) | Opn4tauLacZ | Hattar et al., 2002 | Jax: 021153 RRID:MGI:5520170 |
| Genetic reagent (Mus musculus) | Gnat1-/- | PMID: 11095744 | |
| Genetic reagent (Mus musculus) | Gnat2Cpfl3 | PMID: 17065522 | Jax: 006795 |
| Genetic reagent (Mus musculus) | Opn4Cre | Ecker et al., 2010 | RRID:MGI:5285910 |
| Genetic reagent (Mus musculus) | Brn3bzDTA | Chen et al., 2011 | RRID:MGI:5285910 |
| Genetic reagent (Mus musculus) | Brn3bCre | PMID: 24608965 | RRID:IMSR_JAX:030357 |
| Antibody | anti-OPN4 (rabbit polyclonal) | Advanced Targeting Systems | AB-N38 (1:1000) RRID:AB_1608077 |
| Antibody | AlexaFluor 488, anti-rabbit (goat polyclonal) | Life Technologies | A-11008 (1:1000) RRID:AB_143165 |
| Viral reagent | AAV2-hSyn-DIO-hM3Dq-mCherry | UNC Vector Core | |
| Chemical compound, drug | Clozpine-N-oxide | Sigma | |
| Software | R 3.5.2 | https://cran.r-project.org/ | |
| Software | Graphpad Prism 7.0 | https://www.graphpad.com/scientific-software/prism/ |
Additional files
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Transparent reporting form
- https://doi.org/10.7554/eLife.44358.021
