Variation of connectivity across exemplar sensory and associative thalamocortical loops in the mouse
- McGovern Institute for Brain Research, United States
- Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, United States
- Department of Anatomy and Neuroscience, School of Medicine, Autónoma de Madrid University, Spain
Figures
Figure 1 with 2 supplements
Structural differences between MD→PL and MGB→A1 microcircuits are reflected in functional divergence.
(A) Left: Experimental schematic of combined MD and PL recordings with optical activation of MD using Stabilized Step Function Opsins (SSFO). Right: Anatomical expression of AAV-SSFO-GFP in MD. …
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Figure 1—source code 1
MD PL firing rates.
- https://cdn.elifesciences.org/articles/62554/elife-62554-fig1-code1-v2.zip
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Figure 1—source code 2
MGB A1 firing rates.
- https://cdn.elifesciences.org/articles/62554/elife-62554-fig1-code2-v2.zip
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Figure 1—source code 3
MATLAB Script to calculate spike rate changes .
- https://cdn.elifesciences.org/articles/62554/elife-62554-fig1-code3-v2.zip
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Figure 1—source data 1
Change in firing rates normalised to baseline firing.
- https://cdn.elifesciences.org/articles/62554/elife-62554-fig1-data1-v2.xlsx
Figure 1—figure supplement 1
Sorting of units into regular spiking (RS) and fast spiking (FS) neurons.
(A) Recorded units were separated into regular spiking (n = 1372, green) and fast spiking (n = 678, red) cells, via k-means clustering on three feature dimensions: half trough time, peak to trough …
Figure 1—figure supplement 2
Changes in spike rates of RS and FS neurons recorded in the PL or A1 separated by animals.
(A) Proportional change in spike rate of RS neurons recorded in the PL or A1 produced by SSFO activation of the MD (blue) or MGB (orange) separated by animals. Across animals, a decrease in spike …
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Figure 1—figure supplement 2—source data 1
Change in RS and FS firing rates normalized to baseline separated by individual animals.
- https://cdn.elifesciences.org/articles/62554/elife-62554-fig1-figsupp2-data1-v2.xlsx
Figure 2 with 1 supplement
MD→PL axons establish smaller synaptic terminals than MGB→A1 projections.
(A) Schematic diagram of the iontophoretic injection protocol (left) and images of the center of a representative BDA deposit in the lateral MD (right) Scale bar = 500 µm. (B) Low magnification …
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Figure 2—source data 1
Maximal projection areas of BDA labeled boutons.
- https://cdn.elifesciences.org/articles/62554/elife-62554-fig2-data1-v2.xlsx
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Figure 2—source data 2
p-values table for KS test and Dunn's corrected multiple comparisons post Kruskal Wallis test in Figure 2G, H.
- https://cdn.elifesciences.org/articles/62554/elife-62554-fig2-data2-v2.xlsx
Figure 2—figure supplement 1
Images of BDA deposits in lateral MD and ventral MGB.
Counterstained with Cytochrome oxidase or thionin (case MDL #3). Scale bar = 500 µm.
Figure 3 with 3 supplements
MD→PL projections contact a larger proportion of PV+ inhibitory neurons compared to MGB→A1 projections.
(A) Schematics of trans-synaptic anterograde labeling strategies for output targets of mediodorsal (top) and geniculate (bottom) thalamocortical cells. (B) Representative image of PL (top) and A1 …
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Figure 3—source data 1
Laminar distribution of transsynaptic anterograde labeling and overlap with cortical PV+ neurons .
- https://cdn.elifesciences.org/articles/62554/elife-62554-fig3-data1-v2.xlsx
Figure 3—figure supplement 1
Schematics and injection sites of AAV1 mediated anterograde tracing.
(A) Schematic of anterograde tracing protocol of MD output neurons to PL. (B) Injection site of anterograde AAV1 expressing cre in MD marked by Hoescht333342. Scale bar = 200 µm. (C) Schematic of …
Figure 3—figure supplement 2
Anterograde transsynaptic labeling distributions in PL and A1.
Distribution of the number of mCherry+ neurons per section labeled by the anterograde AAV1-cre shows no significant difference between PL neurons labeled by the MD (blue) and A1 neurons labeled by …
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Figure 3—figure supplement 2—source data 1
Quantification of number of transynaptic anterograde neurons in PL vs A1.
- https://cdn.elifesciences.org/articles/62554/elife-62554-fig3-figsupp2-data1-v2.xlsx
Figure 3—figure supplement 3
Immunolabeled PV positive neuron distribution in PL and A1.
Distribution of immunohistochemically labeled parvalbumin positive neurons in the PL (blue) versus A1 (orange) reveal no significant difference between the two cortices. (n.s., Mann Whitney U test). …
Figure 4 with 1 supplement
MGB→A1 neurons innervate excitatory cortical neurons with larger synaptic terminals compared to MD→PL neurons.
(A) Schematic of synaptic labeling mechanism using PV-Cre driver lines or CamKII-Cre virus to label inhibitory or excitatory synapses, respectively. (B) High magnification confocal image of a …
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Figure 4—source data 1
Synaptic counts and volumes of mGRASP labeled synapses.
- https://cdn.elifesciences.org/articles/62554/elife-62554-fig4-data1-v2.xlsx
Figure 4—figure supplement 1
Overlap of mGRASP labeled synapses with vGlut1 and vGlut2.
(A) Representative image of a post-mGRASP labeled pyramidal neuron (magenta) in the PL with, MD→PL synapses labeled by mGRASP in yellow (left), immunohistochemical labelling of Vglut2+ synapses of …
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Figure 4—figure supplement 1—source data 1
Quantification of overlap of mGRASP labeled synapses with vGluT1 and vGluT2.
- https://cdn.elifesciences.org/articles/62554/elife-62554-fig4-figsupp1-data1-v2.xlsx
Figure 5 with 3 supplements
Detailed map of retrograde inputs to geniculate microcircuit compared to mediodorsal microcircuit (Anterior half, AP 2.60 mm to −0.80 mm).
(A–D) Representative confocal images of monosynaptic inputs MD→PL neurons (left) and MGB→A1 neurons (right) across the rostro-caudal axis illustrate distinct input patterns in each microcircuit. …
Figure 5—figure supplement 1
Monosynaptic retrograde tracing from MD→PL and MGB→A1 neurons.
(A) Schematic of monosynaptic input tracing protocol of MD→PL neurons. (B) Top-left: Injection site of retrograde AAV expressing cre in PL marked by Hoescht333342. Top-right: Helper virus expression …
Figure 5—figure supplement 2
Distribution of starters for monosynaptic retrograde tracing from MD→PL and MGB→A1 neurons.
(A) Schematic of starter population distribution across individual animals in the MD (B) Fraction of starter neurons in the thalamus show predominant expression in the lateral MD compared to the …
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Figure 5—figure supplement 2—source data 1
Starter counts for monosynaptic retrograde tracing with rabies viruses.
- https://cdn.elifesciences.org/articles/62554/elife-62554-fig5-figsupp2-data1-v2.xlsx
Figure 5—figure supplement 3
Quantitative summary of retrograde monosynaptic input tracing from MD→PL and MGB→A1 neurons.
(A, B) Significant inputs (>0.5% total input) to MD→PL (A) and MGB→A1 (B) thalamic neurons measured from different brain regions. (c, d) Sagittal schematic of inputs to MD→PL (C) and MGB→A1 (D). …
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Figure 5—figure supplement 3—source data 1
Retrograde inputs to MD vs MGB expressed as a percentage of total inputs.
- https://cdn.elifesciences.org/articles/62554/elife-62554-fig5-figsupp3-data1-v2.xlsx
Figure 6
Detailed map of retrograde inputs to geniculate microcircuit compared to mediodorsal microcircuit (Rostral half, AP 2.60 mm to −0.80 mm).
(A–E) Representative confocal images of monosynaptic inputs to inputs MD→PL neurons (left) and MGB→A1 neurons (right) across the rostro-caudal axis illustrate distinct input patterns in each …
Figure 7 with 1 supplement
Representation and quantification of inputs to the mediodorsal microcircuit compared to the geniculate microcircuit.
(A) Representative images of major cortical inputs to MD→PL loop (left) versus MGB→A1 (right). Scale bars = 200 µm. (B) Quantification of input to starter ratios for each major cortical input region …
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Figure 7—source data 1
Retrograde monosynaptic labeling input counts, input to starter ratios and layerwise distribution of inputs.
- https://cdn.elifesciences.org/articles/62554/elife-62554-fig7-data1-v2.xlsx
Figure 7—figure supplement 1
Mid brain to starter ratios and TRN inputs to MD→PL and MGB→A1 neurons.
(A) Left: Representative confocal image of the major midbrain input to MD→PL neurons (top), and MGB→A1 neurons (bottom). Right: Quantification of input to starter ratio reveals higher number of …
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Antibody | Anti-GFP antibody (chicken polyclonal) | Aves Labs | GFP1010 RRID:AB_2307313 | (1:1000) |
| Antibody | Alexa Fluor 488 goat anti-chicken IgG (goat polyclonal) | Thermo Fisher Scientific | A32931 RRID:AB_2762843 | (1:500) |
| Antibody | Rabbit anti-PV (rabbit polyclonal) | Swant | PV-27 RRID:AB_2631173 | (1:1000) |
| Chemical compound, drug | BDA (10 KDa) | Thermo Fisher Scientific | D1956 | 3% solution |
| Chemical compound, drug | Hoechst 33342 | Thermo Fisher Scientific | H3570 | 1:1000 solution |
| Software, algorithm | ImageJ | NIH | ||
| Software, algorithm | Imaris | Oxford Instruments | Version 9.3.2 | |
| Software, algorithm | Prism 8 | Graphpad | Version 8.0 | |
| Other | EnvA-RVdG-mCherry | Gift; Dr. Ian Wickersham, MIT Chatterjee et al., 2018 | Rabies virus expressing mCherry | |
| Other | AAV1-TREtight-B19G | Gift; Dr. Ian Wickersham, MIT Chatterjee et al., 2018 | AAV expressing B19 variant of G protein | |
| Other | AAV1-syn-FLEX-TA-TVA-GFP | Gift; Dr. Ian Wickersham, MIT Chatterjee et al., 2018 | AAV expressing cre dependent TVA, tTA, and GFP | |
| Other | AAV2/8-CAG-pre-mGRASP-mCerulean | Gift; Dr. Michael Baratta, Univ. of Colorado | N.A. | AAV expressing pre-mGRASP and mCerulean |
| Other | AAV2/8.CAG.Jx-rev.post-mGRASP-2A-dTomato | Gift; Dr. Michael Baratta, Univ. of Colorado | N.A. | AAV expressing cre dependent post-mGRASP and tdTomato |
| Other | AAVrg-hSyn-Cre-WPRE-hGH (retrograde) | Addgene Vector Core | Lot#: 105553-AAVrg | AAV expressing cre recombinase |
| Other | AAV1-hSyn-Cre-WPRE-hGH | Addgene Vector Core | Lot#: 105553-AAV1 | AAV expressing cre recombinase |
| Other | AAV1-pCAG-FLEX-tdTomato-WPRE | Addgene Vector Core | Lot# 51503 | AAV expressing cre dependent tdTomato |
| Other | AAV1-CamKIIa-SSFO-GFP | UNC vector core | N.A. | AAV expressing SSFO and GFP under CamKII promoter |
| Other | pENN-AAV-CamKII-0.4.-Cre-SV40 | Addgene Vector Core | Lot#: 105540 | AAV expressing cre under CamKII promoter |
