1. Neuroscience

Emergence of non-canonical parvalbumin-containing interneurons in hippocampus of a murine model of type I lissencephaly

  1. Tyler G Ekins
  2. Vivek Mahadevan
  3. Yajun Zhang
  4. James A D'Amour
  5. Gülcan Akgül
  6. Timothy J Petros
  7. Chris J McBain  Is a corresponding author
  1. Program in Developmental Neurobiology, Eunice Kennedy-Shriver National Institute of Child Health and Human Development, National Institutes of Health, United States
  2. NIH-Brown University Graduate Partnership Program, United States
  3. Postdoctoral Research Associate Training Program, National Institute of General Medical Sciences, United States
https://doi.org/10.7554/eLife.62373
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Cite this article
  1. Tyler G Ekins
  2. Vivek Mahadevan
  3. Yajun Zhang
  4. James A D'Amour
  5. Gülcan Akgül
  6. Timothy J Petros
  7. Chris J McBain
(2020)
Emergence of non-canonical parvalbumin-containing interneurons in hippocampus of a murine model of type I lissencephaly
eLife 9:e62373.
https://doi.org/10.7554/eLife.62373
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11 figures, 2 tables and 4 additional files

Figures

Figure 1
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Cell-autonomous and non-autonomous effects of Pafah1b1 heterozygous loss on radial migration.

(A) Images of DAPI-stained hippocampus of WT, GlobalLis (global mutation), NkxLis (IN-specific mutation), and EmxLis (PC-specific mutation) mice. (B) Images of PV- immunostaining in WT, GlobalLis, …

Figure 1—source data 1

PV+INT density and % per layer.

https://cdn.elifesciences.org/articles/62373/elife-62373-fig1-data1-v2.xlsx
Figure 2 with 1 supplement
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Morphological reconstructions and firing profiles of WT and GlobalLis PV+INTs.

(A) Examples of WT PV+INTs with reconstructed cells on top (dendrite in black, axon in blue), polar histograms of dendrite and axonal arbor orientation in the middle and firing traces on bottom. …

Figure 2—source data 1

Axon and dendrite polar histograms.

https://cdn.elifesciences.org/articles/62373/elife-62373-fig2-data1-v2.xlsx
Figure 2—figure supplement 1
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PV-TdTomato/PV-IHC colocalization.

GlobalLis mutant hippocampus. Top image displays PV-IHC, middle displays PV-TdTomato, bottom displays superimposition, in a representative GlobalLis hippocampus. There is high level of …

Figure 2—figure supplement 1—source data 1

PV-TdTomato/PV-IHC co-localization.

https://cdn.elifesciences.org/articles/62373/elife-62373-fig2-figsupp1-data1-v2.xlsx
Figure 3 with 1 supplement
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WT PV+INTs consist of two physiological subtypes: FS and NFS cells.

(A) Unbiased cluster analysis dendrogram displays 83 PV+INTs sorted two clusters that represent fast-spiking (FS) and non-fast-spiking (NFS) cells. Inset indicates the optimal number of clusters, …

Figure 3—figure supplement 1
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Cluster analysis and morphophysiology of EmxLis PV+INTs.

(A) Cluster plot (left) and dendrogram (right) of 38 EmxLis PV+INTs. The dendrogram inset indicates two optimal clusters which correspond to FS and NFS cells. (B) Due to similar sorting, WT and …

Figure 4 with 4 supplements
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GlobalLis PV+INTs consist of three physiological subtypes: FS, IS, and NFS cells.

(A) Unbiased cluster analysis indicates emergence of an additional cluster in GlobalLis CA1, displayed in the dendrogram of 45 PV+INTs. Inset indicates optimal detection of three clusters (FS, IS, …

Figure 4—figure supplement 1
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Cluster analysis and morphophysiology of NkxLis PV+INTs.

(A) Cluster plot (left) and dendrogram (right) of 41 NkxLis PV+INTs. The dendrogram inset indicates three optimal clusters which correspond to FS and NFS cells. (B) Due to similar sorting, as well …

Figure 4—figure supplement 2
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Comparison of K-means and Ward clustering.

A Ward Test was performed, and the resulting clusters were compared to results of the original K-means clustering (used in previous figures). The numbers at the bottoms of the column and ends of the …

Figure 4—figure supplement 3
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Sholl analysis of PV+INT physiological subtypes.

( A) Reconstructions of FS (left), IS (middle) and NFS (right) PV+INTs. Concentric circles (superimposed in black) with a radius interval of 50 µm were used for Sholl analysis. (B) Sholl analysis of …

Figure 4—figure supplement 3—source data 1

Sholl analysis of PV+INTs.

https://cdn.elifesciences.org/articles/62373/elife-62373-fig4-figsupp3-data1-v2.xlsx
Figure 4—figure supplement 4
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Membrane and firing properties of PV+INT physiological subtypes.

Frequency distributions of physiological and morphological properties PV+INTs. FS cells are shown in blue, IS cells in purple, NFS cells in pink. The full color circles connecting lines display the …

Figure 5 with 1 supplement
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Connectivity and microcircuitry of PV+INTs.

(A) (i) Examples of paired whole-cell recordings between synaptically connected presynaptic PV+INTs (top traces) and postsynaptic CA1 PCs (bottom traces). 10 individual traces (grey) and an averaged …

Figure 5—source data 1

PV+INT unitary transmission properties.

https://cdn.elifesciences.org/articles/62373/elife-62373-fig5-data1-v2.xlsx
Figure 5—figure supplement 1
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Analysis of excitatory input to PV+INTs.

Statistical analysis of properties of spontaneous excitatory post-synaptic currents (sEPSCs) on PV+INTs. For statistical analysis *p<0.05, **p<0.01.

Figure 5—figure supplement 1—source data 1

sEPSC properties of PV+INTs.

https://cdn.elifesciences.org/articles/62373/elife-62373-fig5-figsupp1-data1-v2.xlsx
Figure 6
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Depolarization block in PV+INTs.

(A) Maximum firing frequencies and depolarizing block in subtypes of PV+INTs. Top left shows FS cell which showed no reduction in firing frequency after current injection of 1500 pA. Top right …

Figure 6—source data 1

Current inducing depolarization block.

https://cdn.elifesciences.org/articles/62373/elife-62373-fig6-data1-v2.xlsx
Figure 7 with 2 supplements
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Examination of Pafah1b haploinsufficiency on PV+INTs using single-nucleus RNA-seq.

(A) Overview of the experimental workflow. (Bi) Uniform Manifold Approximation and Projection (UMAP) dimensional reduction of single-nuclei transcriptomes of hippocampal PV+INTs, highlighting …

Figure 7—figure supplement 1
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Integrated analyses of single-cell/nucleus transcriptomes from PV+INTs from Pafah1b1+/+, Pafah1b1+/-and reference datasets.

(A) UMAP plots of the PV+INTs from Pafah1b1+/+ and Pafah1b1+/- cortical and hippocampal MGE-derived interneurons and pan-GABAergic Allen Institute datasets indicating robust alignments of the PV+INT …

Figure 7—figure supplement 2
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Annotation of PV+INT subtypes based on discrete marker gene expressions.

(A) Single cell heatmap showing scaled expression values for the top 10 transcriptomic markers in each of the PV+INTs clusters across Pafah1b1+/+, Pafah1b1+/- and reference datasets. (B) UMAP plots …

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Tables

Table 1
Membrane, firing and morphological properties of PV+INTs.
WT
FS
67 cells		GlobalLis
FS
20 cells		NkxLis
FS
12 cells		EmxLis
FS
30 cells		WT
IS
0 cells		GlobalLis
IS
21 cells		NkxLis
IS
21 cells		EmxLis
IS
0 cells		WT
NFS
16 cells		GlobalLis
NFS
4 cells		NkxLis
NFS
8 cells		EmxLis
NFS
8 cells
Input resistance (MΩ)82.5 ± 3.663.5 ± 4.261.1 ± 6.375.0 ± 4.7N/a88.5 ± 4.985.5 ± 7.0N/a143.1 ± 11.9132.0 ± 14.4200.7 ± 17.0173.2 ± 9.0
Rheobase (pA)364.2 ± 17.3457.5 ± 22.2533.3 ± 42.3463.3 ± 24.8n/a340.5 ± 20.0342.9 ± 21.1n/a153.1 ± 14.8162.5 ± 20.0125.0 ± 13.4143.8 ± 11.3
Firing Freq 2x Threshold (Hz)137.6 ± 4.7168.5 ± 11.2173.2 ± 14.8155.4 ± 7.4n/a106.2 ± 4.895.6 ± 3.5n/a66.0 ± 4.557.0 ± 11.672.3 ± 4.458.0 ± 3.5
Adaptation Ratio 2x Threshold0.79 ± 0.020.80 ± 0.030.72 ± 0.040.71 ± 0.02n/a0.71 ± 0.030.65 ± 0.03n/a0.67 ± 0.030.79 ± 0.050.79 ± 0.060.52 ± 0.05
Firing Freq 3x Threshold (Hz)170.9 ± 5.1216.5 ± 11.5213.0 ± 15.3195.7 ± 10.3n/a130.2 ± 6.0131.1 ± 5.1n/a96.1 ± 4.674.5 ± 15.1100.8 ± 5.777.8 ± 6.9
Adaptation Ratio 3x Threshold0.78 ± 0.010.82 ± 0.030.71 ± 0.050.71 ± 0.02n/a0.67 ± 0.020.57 ± 0.03n/a0.65 ± 0.030.64 ± 0.100.73 ± 0.050.47 ± 0.05
AP Threshold (mV)−39.9 ± 0.7−41.2 ± 1.2−39.9 ± 1.3−37.8 ± 1.1n/a−40.4 ± 1.1−35.9 ± 1.2n/a−40.6 ± 1.2−38.6 ± 2.8−36.8 ± 1.2−36.8 ± 1.3
AP Half-width (ms)0.42 ± 0.010.34 ± 0.010.35 ± 0.010.36 ± 0.01n/a0.48 ± 0.010.44 ± 0.01n/a0.59 ± 0.020.64 ± 0.070.56 ± 0.040.58 ± 0.04
AP Amplitude (mV)61.1 ± 1.061.2 ± 2.755.8 ± 3.057.5 ± 2.0n/a62.1 ± 2.353.6 ± 2.0n/a65.4 ± 3.264.8 ± 7.055.1 ± 5.764.9 ± 4.8
AP Max Rise Slope (mV/ms)259.9 ± 5.2248.3 ± 13.5268.6 ± 11.7217.0 ± 12.1n/a261.4 ± 10.7231.0 ± 10.3n/a228.6 ± 11.5210.3 ± 25.0197.9 ± 18.3241.0 ± 26.0
AP Max Decay Slope (mV/ms)−188.7 ± 5.5−176.0 ± 14.3−201.6 ± 9.1−211.4 ± 12.6n/a−198.1 ± 12.6−149.2 ± 7.1n/a−136.5 ± 11.5−131.3 ± 20.6−119.2 ± 14.7−125.4 ± 16.8
AHP Amplitude (mV)−16.8 ± 0.6−17.5 ± 0.9−15.3 ± 0.9−16.7 ± 0.7n/a−16.3 ± 0.8−16.0 ± 1.0n/a−15.7 ± 1.2−16.9 ± 1.1−17.0 ± 1.2−11.3 ± 1.7
Membrane Time Constant (ms)8.5 ± 0.57.5 ± 0.46.7 ± 0.48.6 ± 0.4n/a8.7 ± 0.48.5 ± 0.7n/a10.9 ± 0.612.9 ± 0.614.8 ± 1.617.1 ± 1.0
Membrane Capacitance (pF)104.4 ± 5.0117.7 ± 6.6116.8 ± 18.6123.9 ± 7.7n/a103.7 ± 8.6105.7 ± 8.3n/a91.3 ± 6.488.6 ± 6.473.8 ± 4.897.0 ± 4.3
Sag Index0.90 ± 0.010.91 ± 0.010.95 ± 0.010.92 ± 0.01n/a0.90 ± 0.010.91 ± 0.01n/a0.8 ± 0.030.82 ± 0.040.86 ± 0.020.85 ± 0.02
Total Sholl Intersections (Dendrite)48 ± 7
(19 cells)		65 ± 6
(6 cells)		43 ± 8
(8 cells)		56 ± 7
(11 cells)		n/a43 ± 7
(7 cells)		61 ± 11
(8 cells)		n/a26 ± 4
(10 cells)		28 ± 8
(2 cells)		34 ± 11
(3 cells)		38 ± 10
(4 cells)
Total Sholl Intersections
(Axon)		222 ± 22271 ± 44206 ± 44324 ± 51n/a224 ± 56224 ± 41n/a131 ± 15133 ± 397 ± 2195 ± 26
Postnatal age at time of recording (days)29 ± 126 ± 129 ± 129 ± 2n/a26 ± 127 ± 1n/a28 ± 125 ± 327 ± 128 ± 3
Table 1—source data 1

Morphophysiological properties of PV+INTs.

https://cdn.elifesciences.org/articles/62373/elife-62373-table1-data1-v2.xlsx
Table 2
Unitary transmission properties by PV+INT subtype.
WT
FS.BC
9 connections		WT
FS.DTC
7 connections		WT
NFS.BC
3 connections		GlobalLis
FS
connections		GlobalLis
IS
connections
euIPSC potency (pA)171 ± 7426 ± 556 ± 1071 ± 1656 ± 20
euIPSC Amplitude (pA)171 ± 7424 ± 449 ± 1566 ± 1548 ± 20
Failure probability0.01 ± 0.010.11 ± 0.050.23 ± 0.190.03 ± 0.020.17 ± 0.06
Latency (ms)1.2 ± 0.11.1 ± 0.12.4 ± 0.30.9 ± 0.11.6 ± 0.1
Decay time constant (ms)6.6 ± 0.57.0 ± 0.85.4 ± 0.46.5 ± 0.77.7 ± 1.1
Postnatal age at time of recording (days)35 ± 330 ± 427 ± 130 ± 127 ± 2
Table 2—source data 1

PV+INT euIPSC properties.

https://cdn.elifesciences.org/articles/62373/elife-62373-table2-data1-v2.xlsx

Additional files

Supplementary file 1

Transcriptional impairments triggered by Pafah1b1 haploinsufficiency in PV+INT subtypes.

https://cdn.elifesciences.org/articles/62373/elife-62373-supp1-v2.xlsx
Supplementary file 2

Ingenuity Pathway Analysis (IPA) analysis of differentially expressed genes.

https://cdn.elifesciences.org/articles/62373/elife-62373-supp2-v2.xlsx
Supplementary file 3

Genetic and biochemical interactions with Pafah1b1.

https://cdn.elifesciences.org/articles/62373/elife-62373-supp3-v2.xlsx
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