Cellular birthdate predicts laminar and regional cholinergic projection topography in the forebrain
- Neuroscience Institute, New York University, United States
- Department of Neurobiology, Harvard Medical School, United States
- Stanley Center for Psychiatric Research, Broad Institute, United States
- Department of Neurosurgery, Massachusetts General Hospital and Harvard Medical School, United States
- Northeastern University, United States
Figures
Figure 1 with 3 supplements
Retrograde labeling of layer-specific cholinergic projections in the somatosensory cortex.
See also Figure 1—figure supplements 1, 2 and 3. (A) Genetic and viral strategy for specific labeling of cholinergic efferents to superficial or deep layers of the somatosensory cortex. (B) …
Figure 1—figure supplement 1
Precise and efficient recombination mediated by the Chat-ires-Flpo driver.
(A–D) Anti-ChAT immunohistochemistry (IHC; green) performed on coronal tissue sections (20 μm thick) from P17 ChatFlpO; Dlx6aCre (pan-ventral forebrain driver); Rosa26Ai65 brains. ChAT+ cells …
Figure 1—figure supplement 2
Complete reconstruction of a L1-targeting cholinergic projection neuron.
Left top, front view. Left middle, top view. Left bottom, side view. Right, 3D view.
Figure 1—figure supplement 3
Complete reconstruction of a L5/6-targeting cholinergic projection neuron.
Left top, front view. Left middle, top view. Left bottom, side view. Right, 3D view.
Figure 2
Spatial embryonic origin does not predict projection topography of cholinergic neurons.
(A) Genetic strategy for fate mapping cells from the medial ganglionic eminence (MGE) or preoptic area (POA) based on differential gene expression. (B) Fate mapped cells in the P30 medial septum …
Figure 3 with 1 supplement
Cellular birthdate predicts layer-specific cholinergic projection topography in the primary somatosensory cortex.
See also Figure 3—figure supplement 1. (A) Experimental design for tamoxifen (Tm) induction of CreER activity for neuronal birthdating. (B) Example of E11 tamoxifen-birthdated cholinergic neurons at …
Figure 3—figure supplement 1
Birthdated cholinergic neuron projections to the primary motor and primary visual cortices.
(A) Quantification of overall primary motor cortex (M1) innervation at P30 for cholinergic neurons born at each timepoint (one-way ANOVA (F(3,9) = 4.458), p=0.0352). (B) Quantification of …
Figure 4
Birthdate predicts cholinergic topography in the hippocampus, amygdala, and mPFC.
(A) Examples of birthdated cholinergic neuron projections to the medial prefrontal cortex (mPFC). (B) Examples of birthdated cholinergic neuron projections to the hippocampus at P30. (C) Examples of …
Figure 5
Cell body distribution of birthdated cholinergic neurons.
(A) Cell body locations in the basal forebrain at P30 for cholinergic neurons born at each timepoint labeled by tamoxifen induction of CreER. (B) Quantification of cell body distribution (not …
Figure 6
Cholinergic axons traverse distinct routes to their projection targets based on birthdate.
(A) Routes cholinergic axons take to the cortex from the basal forebrain. (B) Proportion of fibers in each pathway for brains with birthdated cholinergic neurons at P30. Connected lines represent a …
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (canine adenovirus) | Cav-2::Cre | Plateforme de Vectorologie de Montpellier | CAV Cre | |
| Genetic reagent (Mus musculus) | ChatFlpo | This paper | N/A | Submitted to Jax, further information about generation of this line in Materials and methods below. |
| Genetic reagent (Mus musculus) | CMVCre | Jax | RRID:IMSR_JAX:006054 | |
| Genetic reagent (Mus musculus) | Ascl1CreER | Jax | RRID:IMSR_JAX:012882 | |
| Genetic reagent (Mus musculus) | Lhx6iCre | Jax | RRID:IMSR_JAX026555 | |
| Genetic reagent (Mus musculus) | ShheGFP-Cre | Jax | RRID:IMSR_JAX005622 | |
| Genetic reagent (Mus musculus) | Rosa26Ai65 | Jax | RRID:IMSR_JAX021875 | |
| Antibody | Anti-DsRed (rabbit polyclonal) | Clontech | RRID:AB_10013483 | (1:1000) |
| Antibody | Anti-ChAT (goat polyclonal) | Millipore | RRID:AB_2079751 | (1:250) |
| Antibody | Anti-rabbit IgG Alexa Fluor 594 (donkey polyclonal) | Thermo Fisher | RRID:AB_141637 | (1:1000) |
| Antibody | Anti-goat IgG Alexa Fluor 594 (donkey polyclonal) | Thermo Fisher | RRID:AB_2534105 | (1:1000) |
| Commercial assay or kit | Click-iT EdU kit for imaging Alexa Fluor 488 | Thermo Fisher | C10337 | |
| Chemical compound, drug | 5-Ethynyl-2´- deoxyuridine (EdU) | Invitrogen | A10044 | (50 μg per 1 g body weight) |
| Software, algorithm | Neurolucida 360 | MBF Biosciences | Neurolucida 360 2019 and 2020 | |
| Other | DAPI stain | Invitrogen | D1306 | (1 µg/mL) |
Additional files
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Supplementary file 1
ANOVA summary tables and post-hoc test information.
- https://cdn.elifesciences.org/articles/63249/elife-63249-supp1-v1.xlsx
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Transparent reporting form
- https://cdn.elifesciences.org/articles/63249/elife-63249-transrepform-v1.pdf
