Activity dynamics of amygdala GABAergic neurons during cataplexy of narcolepsy

  1. Ying Sun
  2. Carlos Blanco-Centurion
  3. Emmaline Bendell
  4. Aurelio Vidal-Ortiz
  5. Siwei Luo
  6. Meng Liu  Is a corresponding author
  1. Medical University of South Carolina, United States
6 figures, 4 videos, 3 tables and 1 additional file

Figures

Histology results.

(A) Immunostaining demonstrating abundant orexin immunoreactive neurons and fibers in the LH of the control mice (n = 8). (B) orexin immunoreactive somata and fibers were utterly absent in LH of the …

https://doi.org/10.7554/eLife.48311.002
Figure 1—source data 1

Cell counts of neurons expressing GCaMP6 and VGAT in the amygdala of one set of coronal sections from the narcoleptic mice used for calcium data analysis (n = 5).

Percentages inside the parenthesis in column (A) were calculated as of total GCaMP6 expressing neurons in the amygdala structure. More than 80% of GCaMP6 expressing neurons were located in CeA. About 95% of them also contained VGAT. CeA: central nucleus of the amygdala, BLA: basolateral amygdala, BMA: basomedial amygdala.

https://doi.org/10.7554/eLife.48311.003
Live Ca2+ activity recording and cell classification.

(A) Map of 44 recorded cells from narcoleptic mouse LR02082418. (B) A serial figures showing (top to bottom) EEG spectrogram, EMG, brain states, cell traces of a total of 44 cells, cell traces of …

https://doi.org/10.7554/eLife.48311.004
Activities of sorted neurons during sleep/wake cycle and cataplexy.

(A–C) Neuronal activity graphs of 28 HAW-ON cells, 19 EC-ON cells and 17 unscored cells in the narcoleptic mouse LR02082418. Most of the unscored cells displayed significantly elevated activities …

https://doi.org/10.7554/eLife.48311.006
Figure 3—source data 1

Z-score data for transition map.

https://doi.org/10.7554/eLife.48311.007
Figure 4 with 1 supplement
Hyperactivity of amygdala GABAergic neurons before and during emotion-induced cataplexy.

(A–D), heat maps of Ca2+ fluorescent intensity changes before and after coyote urine exposure (red arrows show the starting time of exposure) in control mouse LR04101618. (A and B) and narcoleptic …

https://doi.org/10.7554/eLife.48311.011
Figure 4—source data 1

Percentage source data for Figure 4 heat maps and graphs (A–D, I, J).

https://doi.org/10.7554/eLife.48311.013
Figure 4—figure supplement 1
Calcium transient intensity ΔFF (Z-score) graphs (1-4) are incorporated into each heat maps in Figure 4, A-D, respectively, to demonstrate the dramatic intensity increase in the narcoleptic mice after coyote urine exposure.

Legend is the same as Figure 4.

https://doi.org/10.7554/eLife.48311.012
Spatial connectivity maps and graphs.

Colored lines represent a strong positive correlation R value between 0.6 (blue) to 1.0 (red). (A–D): correlation maps of an SC (A), EC (B), HAW (C) and REMS (D) bout from narcoleptic mouse …

https://doi.org/10.7554/eLife.48311.017
Figure 5—source data 1

source data for correlation maps and graph.

https://doi.org/10.7554/eLife.48311.018
Durations of 10 spontaneous (SC) and 14 emotion-induced cataplexy (EC).

The duration difference between SC and EC are insignificant (F(1,22)=3.02, p=0.09).

https://doi.org/10.7554/eLife.48311.019
Figure 6—source data 1

Source data for duration graph.

https://doi.org/10.7554/eLife.48311.020

Videos

Video 1
A sample of raw calcium movies played at 64x speed.
https://doi.org/10.7554/eLife.48311.010
Video 2
Behaviors upon coyote urine exposure from a control mouse (Left) and a narcoleptic mouse (right).
https://doi.org/10.7554/eLife.48311.014
Video 3
Demonstrating the spontaneous cataplexy (SC) bout in Figure 4C, 8x speed.
https://doi.org/10.7554/eLife.48311.015
Video 4
Demonstrating the emotion-induced cataplexy (EC1) bout in Figure 4D, 8x speed.
https://doi.org/10.7554/eLife.48311.016

Tables

Table 1
Number of recorded cells in each mouse.
https://doi.org/10.7554/eLife.48311.008
Mouse IDSexGroupNumbers of cells
LR00062218MaleNarcoleptic27
LR01062918FemaleNarcoleptic35
LR02082418MaleNarcoleptic44
LR03091618MaleNarcoleptic42
LR08012219FemaleNarcoleptic38
LR04101618MaleControl38
LR12011419MaleControl28
LR13022419FemaleControl63
LR14022819FemaleControl35
LR16031319MaleControl43
Table 2
components of HAW-ON and EC-ON neurons in the narcoleptic group.
https://doi.org/10.7554/eLife.48311.009
UnscoredAW-ONREMS-ON REMS/AW-ONNREMS-ON
SC-ON
EC-ONHAW-ON
HAW-ON (112)50 (44.64%)40 (35.71%)13 (11.61%)8 (7.14%)63 (56.25%)------------
EC-ON (74)40 (54.05%)17 (22.97%)17 (22.97%)0------------63 (85.14%)
Key resources table
Reagent type
(species) or resource
DesignationSource or referenceIdentifiersAdditional
information
Genetic reagent (M. musculus)VGAT-CreThe Jackson laboratoryRRID: MGI:5141285PMID:21745644
Genetic reagent (M. musculus)Orexin KOPMID:10481909RRID: IMSR_KOMP:VG11744-1.1-VlcgDr. Masashi Yanagisawa (University of Texas)
Recombinant DNA reagentAAV.Syn.Flex.GCaMP6s.WPRE.SV40http://www.addgene.org/100845/RRID:Addgene_100845PMID:23868258
Originally purchased from university of Pennsylvania gene
therapy core
Antibodyanti-VGAT (Mouse monoclonal)Synaptic SystemCat# 131011, RRID:AB_887872PMID:17444497
(1:500)
Antibodyanti-Orexin (Goat polyclonal)Santa Cruz BiotechnologyCat#: SC-8070,
RRID:AB_653610
PMID:16100511
(1:5000)
Software, algorithmSPSSIBMRRID:SCR_002865
Software, algorithmMatlabMathWorksRRID:SCR_001622

Additional files

Download links