Coordinated cadherin functions sculpt respiratory motor circuit connectivity
- Department of Neurosciences, Case Western Reserve University, United States
- Department of Developmental Neurobiology, St. Jude Children's Research Hospital, United States
- Department of Ophthalmology, University of California, San Francisco, United States
- Max Delbrück Center for Molecular Medicine in the Helmholtz Association, Germany
Figures
Figure 1 with 5 supplements
A combinatorial cadherin code establishes breathing and is required for life.
(a) A combinatorial cadherin code defines phrenic MNs during development. Reproduced from Figure 4c from Vagnozzi et al., 2020. The ventral lateral spinal cord at the level of the PMC (brown circle) …
Figure 1—figure supplement 1
Validation of NMNΔ6910KO mice.
(a) N-cadherin, cdh6, 9, and 10 expression in phrenic motor neurons (MNs) at e13.5 (modified from Vagnozzi et al., 2020). The images shown are taken from the ventral lateral spinal cord at the level …
Figure 1—figure supplement 2
Normal breathing behaviors in 6910KO mice.
(a) Schematic of a representative breath and related respiratory parameters recorded by plethysmography (adapted from emka user manual). (b) 6910KO mice do not show changes in minute ventilation at …
Figure 1—figure supplement 3
Cadherins N, 6, 9, and 10 control diaphragm innervation.
(a) Diaphragm innervation in control, 6910KO, NMNΔ, and NMNΔ6910KO mice. NMNΔ6910KO mice display a reduction in ventral diaphragm innervation (arrows) and arborization complexity at e18.5. Motor …
Figure 1—video 1
Respiratory insufficiency in NMNΔ6910KO mice.
NMNΔ6910KO mice appear cyanotic at birth and gasp for air before perishing shortly after birth.
Figure 1—video 2
Functional neuromuscular junctions (NMJs) in NMNΔ6910KO mice.
Stimulation of the phrenic nerve at e18.5/P0 in NMNΔ6910KO mice can elicit robust diaphragm contractions, demonstrating functional NMJs.
Figure 2 with 1 supplement
N-cadherin predominantly establishes phrenic motor neuron (MN) cell body position.
Analysis of cell body position reveals differential contributions of type I and II cadherins to phrenic motor topography. (a, c, e, g) Phrenic MN cell bodies (yellow, defined by the co-expression of …
Figure 2—figure supplement 1
N-cadherin predominantly establishes phrenic motor neuron (MN) cell body position.
(a) Rostral to caudal distribution of e13.5 phrenic MN cell bodies (yellow, defined by the expression of Scip in green and Isl1/2 in red) in control, 6910KO, NMNΔ, and NMNΔ6910KO mice. While phrenic …
Figure 3 with 1 supplement
Cadherins N, 6, 9, and 10 dictate phrenic motor neuron (MN) dendritic orientation.
(a, c, e, g) DiI injections into the phrenic nerve in control, 6910KO, NMNΔ, and NMNΔ6910KO mice reveal phrenic MN dendrites, which extend in the dorsolateral and ventromedial directions in control …
Figure 3—figure supplement 1
Phrenic motor neuron (MN) dendritic orientation requires cadherins N, 6, 9, and 10.
(a–d) Rostral to caudal extent of phrenic MN dendrites, as revealed by DiI injections into the phrenic nerve. In control and 6910KO mice, dendrites elaborate in dorsolateral to ventromedial …
Figure 4
Coordinated type I and II cadherin signaling drives phrenic motor neuron (MN) activation.
(a) Schematic of brainstem–spinal cord preparation, which displays fictive inspiration after removal of the pons. Suction electrode recordings were taken from the phrenic nerve in the thoracic …
Figure 5 with 1 supplement
Cadherin expression defines the core motor respiratory circuit.
Fluorescence in situ hybridization (FISH) showing expression of Cdh6 (green, a, b), Cdh9 (green, c, d), Cdh10 (green, e, f), and N-cadherin (green, g, h) in the brainstem, at the putative location …
Figure 5—figure supplement 1
Validation of Cdh9iCre mice.
Cre-dependent fluorescent reporter expression (Ai9-Tdtomato, red) in Cdh9iCre mice labels phrenic motor neurons (MNs) at e12.5 (a, identified by Scip expression in green) and the putative rostral …
Figure 6 with 1 supplement
Cadherin signaling is required in Dbx1-derived neurons for robust respiratory output.
(a) B- and γ-catenin are obligate intracellular factors required for cadherin-mediated cell adhesive function. We utilized inactivation of β- and γ-catenin in Dbx1-derived interneurons (Ctnnb1flox/fl…
Figure 6—figure supplement 1
Dbx1-mediated recombination does not target phrenic motor neurons (MNs).
Cre-dependent fluorescent reporter expression (Ai9-Tdtomato, red) in Dbx1Cre mice does not label phrenic MNs (circled) at e13.5 (identified by Isl1/2 and Scip expression in green and blue, …
Figure 7
Coordinated cadherin functions dictate respiratory motor output.
In control animals, phrenic motor neurons (MNs) form a cluster in the cervical, ventral spinal cord and project their dendrites in a stereotypical ventromedial and dorsolateral orientation. This …
