Journal: The Journal of Neuroscience

Article Title: Dorsal Raphe Serotonin Neurons Mediate CO 2 -Induced Arousal from Sleep

doi: 10.1523/JNEUROSCI.2182-17.2018

Figure Lengend Snippet: Stimulation of DRN with acidosis-induced arousal from sleep in Lmx1bf/f but not Lmx1bf/f/p mice. A, Sagittal schematic view of mouse brain demonstrating cannula placement into DRN (green). Medullary raphe depicted in purple for reference. B, EEG and EMG traces depicting lack of arousal from sleep with perfusion of normal aCSF into DRN (left) and arousal from sleep with perfusion of acidified aCSF into DRN (right) in an Lmx1bf/f mouse. Calibration: vertical, 20 μV; horizontal, 20 s. C, EEG and EMG traces depicting lack of arousal from sleep with perfusion of normal (left) or acidified (right) aCSF into DRN of an Lmx1bf/f/p mouse. Calibration as in B. D, Mean arousal latencies for Lmx1bf/f (black) and Lmx1bf/f/p (red) mice treated with normal (control; solid bars) or acidified (acidosis; open bars) aCSF. Data are mean ± SEM. n = 10 per group. *p < 0.001. E, Percentage change in time spent in Wake (left) and NREM (right) for Lmx1bf/f (black) and Lmx1bf/f/p (red) mice while DRN perfused with acidified aCSF (acidosis; open bars) compared with normal aCSF (control; solid bars). n = 10 per group. *p < 0.001. F, Nissl-stained section to verify cannula placement toward DRN. Arrow indicates cannula tract. Scale bar, 100 μm. Aq, Aqueduct of Sylvius. G–I, Bar graphs represent mean effects of perfusion of normal or acidified aCSF into DRN on respiratory frequency (G), tidal volume (H), and minute ventilation (I) for Lmx1bf/f (black) and Lmx1bf/f/p (red) mice during Wake (left) and NREM (right). n = 10 per group.

Article Snippet: The guide cannula was then secured with dental acrylic glue (Lang Dental); and once firmly in place, the electrode wires were soldered to the six-pin EEG/EMG head socket (Pinnacle Technology or Bisco Industries).

Techniques: Staining

Journal: The Journal of Neuroscience

Article Title: Dorsal Raphe Serotonin Neurons Mediate CO 2 -Induced Arousal from Sleep

doi: 10.1523/JNEUROSCI.2182-17.2018

Figure Lengend Snippet: Inactivation of 5-HT neurons through systemic application of 8-OH-DPAT to Htr1aRR mice prevented arousal to CO2. A, EEG and EMG traces depicting arousal response following 7% CO2 challenges 60 min after saline (left) or 8-OH-DPAT (0.5 mg/kg; right) injection (100 μl, s.c.) in a Htr1aRR mouse. Calibration: vertical, 5 μV; horizontal, 30 s. B, Mean latency to arousal following CO2 challenge 60 min after saline (green) or 8-OH-DPAT (purple) injections in Htr1aKO and Htr1aRR mice as indicated. n = 6 per group. *p < 0.05. **p < 0.001.

Article Snippet: The guide cannula was then secured with dental acrylic glue (Lang Dental); and once firmly in place, the electrode wires were soldered to the six-pin EEG/EMG head socket (Pinnacle Technology or Bisco Industries).

Techniques: Injection

Journal: The Journal of Neuroscience

Article Title: Dorsal Raphe Serotonin Neurons Mediate CO 2 -Induced Arousal from Sleep

doi: 10.1523/JNEUROSCI.2182-17.2018

Figure Lengend Snippet: Inactivation of DRN 5-HT neurons through direct application of 8-OH-DPAT to DRN of Htr1aRR mice prevented arousal to CO2. A, EEG and EMG traces depicting arousal response to 7% CO2 during perfusion of DRN with normal aCSF (left) or aCSF containing 8-OH-DPAT (1 mm; right) in an Htr1aRR mouse. Calibration: vertical, 20 μV; horizontal, 20 s. B, Latency to arousal to CO2 challenge during perfusion of DRN with normal aCSF (green) or 8-OH-DPAT (purple) in Htr1aKO and Htr1aRR mice as indicated. n = 6 per group. *p < 0.001.

Article Snippet: The guide cannula was then secured with dental acrylic glue (Lang Dental); and once firmly in place, the electrode wires were soldered to the six-pin EEG/EMG head socket (Pinnacle Technology or Bisco Industries).

Techniques:

Journal: The Journal of Neuroscience

Article Title: Dorsal Raphe Serotonin Neurons Mediate CO 2 -Induced Arousal from Sleep

doi: 10.1523/JNEUROSCI.2182-17.2018

Figure Lengend Snippet: Optogenetic inhibition of DRN 5-HT neurons prevented arousal to CO2. A, Schematic of virus transfection and placement of fiberoptic cannula into DRN (green). Medullary raphe depicted in purple for reference. B, Coronal sections showing virus transfection in the DRN of a Pet1-Cre mouse (top) and lack of expression in a wild-type mouse (bottom). C, EEG and EMG traces depicting arousal from sleep to CO2 without (left) and with (right) laser stimulation in a Pet1-Cre mouse transfected with Arch. Green traces represent time of laser on. Calibration: vertical, 20 μV; horizontal, 60 s. D, Mean arousal latencies in mice expressing Arch (bottom) and in control mice not expressing a functional opsin (top) to room air (open bars) and CO2 (closed bars) exposure without (black) and with (green) laser stimulation. Data are mean ± SEM. n = 6 per group. *p < 0.05. **p < 0.001.

Article Snippet: The guide cannula was then secured with dental acrylic glue (Lang Dental); and once firmly in place, the electrode wires were soldered to the six-pin EEG/EMG head socket (Pinnacle Technology or Bisco Industries).

Techniques: Inhibition, Transfection, Expressing, Functional Assay