TMEM16B regulates anxiety-related behavior and GABAergic neuronal signaling in the central lateral amygdala
- University of California, San Francisco, United States
- Howard Hughes Medical Institute, University of California, San Francisco, United States
- Gladstone Institute of Neurological Disease, United States
Figures
Figure 1
Ano2 Mutant Mice Display Reduced Anxiety-related Behavior and Impaired Pre-pulse Inhibition.
(A) Representative traces of Ano2 KO mice and control (Con) littermates in open field. (B–D) Ano2 mutant mice display reduced anxiety-related behavior in the open field test, shown as time spent at …
Figure 2
Ano2 Knockout Mice Exhibit Reduced Context-independent Cued Fear Expression.
(A) Ano2 KO and control mice exhibited comparable freezing levels during fear acquisition. After habituation for 5 min, freezing levels during fear training with three 30 s 80 dB tones that each was …
Figure 3 with 1 supplement
Ano2 mRNA Expression in Somatostatin-positive GABAergic Neurons in the Central Lateral Amygdala.
(A–C) RNA scope studies reveal that Ano2 is expressed in central lateral amygdala (CeL), but not in basal lateral amygdala (BLA) or central medial amygdala (CeM). Representative images of …
Figure 3—figure supplement 1
Ano2 mRNA Expression Patterns.
(A and B) RNA scope labeling of Ano2 mRNA in lateral septum, CA1, CA3 and DG. (C) No detectable signals are observed in samples from Ano2 KO mutant mice as specificity control.
Figure 4 with 1 supplement
TMEM16B Mediated Ca2+-activated Cl- Currents in Somatostatin-positive Neurons of the Central Lateral Amygdala.
(A) Brain slice of Sst-IRES-Cre and Ai14 reporter mice with fluorescently marked somatostatin-positive (SOM+) neurons in the amygdala. Scale bar, 200 μm. (B and C) Depolarization of SOM+ neurons to …
Figure 4—figure supplement 1
No detectable CaCC in non-SOM+ neurons.
(A) Calcium current in SOM+ neurons induced by 60 ms voltage clamp steps from −40 mV to +10 mV in 10 mV increments. (B and C) Depolarization of SOM+ neurons (B) and non-SOM+ neurons (C) to +10 mV …
Figure 5 with 1 supplement
TMEM16B-CaCC Regulates Action Potential Waveform of Somatostatin-positive Neurons in the Central Lateral Amygdala.
(A) upper panel, representative traces of action potentials in SOM+ neurons from control and Ano2 KO mice in response to 100 pA current pulse lasting for 500 ms. Lower panel, overlay of action …
Figure 5—figure supplement 1
SOM+ Neurons Lacking Ano2 Have Normal Resting Potential, Membrane Capacitance, Input Resistance, Time Constant, Voltage threshold and Afterhyperpolarization.
(A–D) Passive membrane property is measured in SOM+ neurons from control and Ano2 KO mice, including capacitance (A), input resistance (B), membrane time constant (C), resting membrane potential (D),…
Figure 6 with 1 supplement
Altered Inhibitory Neurotransmission in the Central Lateral Amygdala of Ano2 KO mice.
(A) Representative traces of sIPSCs from SOM+ neurons of Ano2 KO and control mice. (B) The SOM+ neurons of Ano2 KO mice showed increased sIPSC frequency and sIPSC amplitude. Mann Whitney test in (B),…
Figure 6—figure supplement 1
Similar eIPSC amplitudes of SOM+ neurons in control and Ano2 knockout mice.
(A) Representative IPSC induced by 100 μm GABA puff from control (indicated as black) and KO (red) mice (puff duration 50 ms, 4–6 psi). The GABA induced IPSC is blocked by picrotoxin (PTX), a GABAA …
Figure 7
Voltage-activated Ca2+ Channel Involvement in mIPSC in Brain Slices With or Without Ano2.
(A) Representative traces of mIPSCs from SOM+ neurons of Ano2 KO and control mice. (B) The SOM+ neurons of Ano2 KO mice showed a decrease of mIPSC frequency, and normal mIPSC amplitude. Mann Whitney …
Figure 8
Mechanistic model for how presynaptic loss of function of TMEM16B, a Ca2+-activated Cl- channel, in central lateral amygdala (CeL) SOM+ GABAergic neurons mediates the electrophysiological and behavioral phenotypes.
(A) Loss of TMEM16B mediated CaCC in SOM+ CeL neurons results in broadening of action potential, thereby increasing presynaptic calcium channel activity and GABA release evoked by spike firing. (B) …
Additional files
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Transparent reporting form
- https://doi.org/10.7554/eLife.47106.014
