Necdin shapes serotonergic development and SERT activity modulating breathing in a mouse model for Prader-Willi syndrome
- Aix Marseille Univ, INSERM, INMED, France
- Karolinska Institute, Sweden
- Université Paris 13, UFR STAPS, France
- Case Western Reserve University, United States
- UPMC Univ Paris 6, Institut du Fer à Moulin, INSERM, France
- Université de Lyon, INSERM, CNRS, France
- Aix Marseille Univ, CNRS, IBDM, France
Figures
Figure 1 with 3 supplements
Necdin expression in 5-HT neurons and alterations of 5-HT neuronal development and activity in Ndn-KO mice.
(A) Scheme adapted from (Hawthorne et al., 2010) representing expression profiles of Necdin (green), Pet1 (blue) and 5-HT (red) throughout embryonic development of 5-HT neurons as soon as the …
Figure 1—figure supplement 1
Necdin expression compared with Pet-1 and 5-HT expression throughout embryonic development and alteration of 5-HT projections in Ndn-KO embryos.
(A–I) Co-expression in WT brainstem of Necdin (B, E, H) with Pet-1 (A) and 5-HT (D,G) at E10.5 (A–C), E12.5 (D–F) and E16.5 (G–I). (J) 5-HT IHC on coronal brainstem sections at E12.5 showing 5-HT …
Figure 1—video 1
Two-photon timelapse video showing somal translocation on organotypic slice cultures of Pet-EYFP neurons in WT embryos (E12.5).
https://doi.org/10.7554/eLife.32640.005
Figure 1—video 2
Two-photon timelapse video showing somal translocation on organotypic slice cultures of Pet-EYFP neurons in Ndn-KO embryos (E12.5).
https://doi.org/10.7554/eLife.32640.006
Figure 2 with 3 supplements
Large punctiform axonal 5-HT staining (5-HT LPAs) results from an increase in SERT expression and activity in Ndn-KO mice.
(A–B) (A) Axonal 5-HT immunoreactivity illustrating 5-HT LPAs in the raphe of WT, Ndn-KO, Slc6a4-KO and Ndn/Slc6a4-DKO neonates (P1). (B) 5-HT LPAs were counted for all different genotypes (n = 3/gen…
Figure 2—figure supplement 1
5-HT metabolism, Tph2 and Slc6a4 transcripts quantification in Ndn-KO mice.
(A–D) 5-HT metabolic analyses from medulla extracted from WT (n = 8) and Ndn-KO (n = 6) mice (at E18.5): (A) 5-HT substrate (L-Trp) (mg per gram of tissue): WT: 85.8 (64.5, 98.5); Ndn-KO: 83.6 …
Figure 2—figure supplement 2
ASP+ uptake in neurons of raphe primary cultures.
(A) 5-HT immunocytochemistry on primary raphe cultures showing positive 5-HT neurons (red). (B) Time lapse illustration of ASP+ fluorescence (black) accumulation into cells bodies and fibers over 5 …
Figure 2—figure supplement 3
Flow diagram of mice used for in vitro and in situ analyses in Figures 1 and 2 and their corresponding supplement figures.
https://doi.org/10.7554/eLife.32640.010
Figure 3 with 2 supplements
Genetic ablation or pharmacologic inhibition of SERT suppresses apnea and rescues central chemoreflex in Ndn-KO mice.
(A) Workflow experiment of constant airflow whole body plethysmography performed in unanaesthetized, unrestrained WT, Ndn-KO, Ndn/Slc6a4-DKO and Ndn-KO+Fluox mice at the age of P15. Ndn-KO and WT …
Figure 3—figure supplement 1
Early life Fluoxetine treatment has only short-term positive effects on Ndn-KO apneas.
(A) Workflow experiment of constant airflow whole body plethysmography performed in unanaesthetized, unrestrained WT and Ndn-KO mice at 0, 15 and 45 days after treatment (DAT). Ndn-KO and WT animals …
Figure 3—figure supplement 2
Early life treatment of Fluoxetine on respiratory apnea in wild-type mice.
(A–B) Plethysmographic recordings of WT mice at 45DAT pre-treated either with (A) 0.9% NaCl (indicated here as WT) or (B) Fluoxetine (indicated here as WT+Fluox) (10 mg/Kg/day) from age P5 to P15 …
Figure 4 with 3 supplements
Alteration of respiratory chemoreflex in Ndn-KO neonates is rescued by Fluoxetine.
(A–F) Effect of hypercapnia on in vivo ventilatory parameters of WT and Ndn-KO neonates. (A) Workflow experiment of constant airflow whole body plethysmography performed in unanaesthetized, …
-
Figure 4—source data 1
Plethysmography data before and after hypercapnia in WT and Ndn-KO mice.
- https://doi.org/10.7554/eLife.32640.018
-
Figure 4—source data 2
Electrophysiology data of rhythmic phrenic bursts frequency during acidosis in WT and Ndn-KO preparations - before and after Fluoxetine treatment.
- https://doi.org/10.7554/eLife.32640.019
-
Figure 4—source data 3
Relates to Figure 4—figure supplements 1 and 2.
Electrophysiology data of rhythmic phrenic bursts frequency during acidosis in WT and Ndn-KO preparations - before and after 8-OHDPAT treatment.
- https://doi.org/10.7554/eLife.32640.020
Figure 4—figure supplement 1
Effect of pre-treatment with the 5-HT1A-R agonist 8OHDPAT on the resting PBf and the PBf response to acidosis in Ndn-KO en bloc brainstem-spinal cord preparations of P0-P1 pups.
(A–C) PBf produced in Ndn-KO en bloc brainstem-spinal cord preparations superfused with (A) neutral aCSF (pH 7.4) and then treated with 8OHDPAT (1 µM) either (B) in neutral aCSF (pH 7.4) or (C) in …
Figure 4—figure supplement 2
Effects of Fluoxetine and of the 5-HT1A-R agonist 8OHDPAT on the resting PBf and the PBf response to acidosis in wild-type medulla preparations.
(A–B) PBf produced in WT en bloc brainstem-spinal cord preparations superfused with (A) neutral aCSF (pH 7.4) and treated with Fluoxetine (20 μM) in aCSF (pH 7.4) or acidified aCSF (pH 7.1), (B) …
Figure 4—figure supplement 3
Flow diagram of mice used for ex vivo and in vivo analyses in Figures 3 and 4 and their corresponding supplement figures.
https://doi.org/10.7554/eLife.32640.017Additional files
-
Transparent reporting form
- https://doi.org/10.7554/eLife.32640.021
