Differential spatiotemporal development of Purkinje cell populations and cerebellum-dependent sensorimotor behaviors
- Department of Neuroscience, Erasmus MC, Netherlands
- Princeton Neuroscience Institute, United States
- Department for Developmental Origins of Disease, University Medical Center Utrecht Brain Center and Wilhelmina Children’s Hospital, Netherlands
Figures
Figure 1 with 5 supplements
Simple and complex spike activity develops differently between ZebrinII-related Purkinje cell populations.
(A1) Schematic illustration of single unit cerebellar extracellular recordings in Purkinje cells of awake head-fixed mice (left). Schematic representation of a cerebellar coronal section and …
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Figure 1—source data 1
Purkinje cell in vivo source data 1.
- https://cdn.elifesciences.org/articles/63668/elife-63668-fig1-data1-v2.xlsx
Figure 1—figure supplement 1
In vivo extracellular recordings of simple and complex spike activity in different ZebrinII-related Purkinje cell populations during development.
(A1) Example trace of a young P12 Purkinje cell recording identified by its hallmark feature, the occurrence of complex spikes (asterisk) and simple spikes (left). Inset depicting the complex spike …
Figure 1—figure supplement 2
Climbing fiber pause is longer than inter-simple spike-interval in the majority of the recorded cells.
(A) Scatterplot of the log mean simple spike-interval versus the log climbing fiber (CF) pause of all cells recorded in four age groups (blue: P12-17; green: P18-29; red: P30-59 and purple: P>60). …
Figure 1—figure supplement 3
In vivo extracellular recordings of simple spike activity distributed by lobules and cerebellar regions during development.
Purkinje cell simple spike firing rate in vivo for Purkinje cells in all lobules at (A1) >P60, (A2) P30-59, (A3) P18-29 and (A4) P12-17. Purkinje cell simple spike firing rate in vivo for (B1) …
Figure 1—figure supplement 4
Overview with color-coded simple spike and complex spike rate for all identified ZebrinII– and ZebrinII+ Purkinje cells.
(A) P12-P17 Purkinje cell recording location with simple spike rate and (B) P12-P17 Purkinje cell recording location with complex spike rate.
© 2007, John Wiley & Sons. The schematic drawing in panels A and B is redrawn from Figure 3 from Sugihara and Quy, 2007b, with permission from John Wiley & Sons, Inc. It is not covered by the CC-BY 4.0 licence and further reproduction of this panel would need permission from the copyright holder.
Figure 1—figure supplement 5
Overview with color-coded simple spike coefficient of variation 2 (CV2) and coefficient of variation (CV) for all identified ZebrinII– and ZebrinII+ cells.
(A) P12-P17 Purkinje cell recording location with simple spike coefficient of variation (CV2) and (B) P12-P17 Purkinje cell recording location with simple spike coefficient of variation (CV).
© 2007, John Wiley & Sons. The schematic drawing in panels A and B is redrawn from Figure 3 from Sugihara and Quy, 2007b with permission from John Wiley & Sons, Inc. It is not covered by the CC-BY 4.0 licence and further reproduction of this panel would need permission from the copyright holder.
Figure 2 with 2 supplements
Intrinsic activity develops differently between lobule III and lobule X Purkinje cell populations.
(A1) Cell-attached recordings in vitro in Slc1a6-EGFP mice that express eGFP in Purkinje cells. Recordings of intrinsic Purkinje cell activity were made in lobules III and X. Purkinje cells were …
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Figure 2—source data 1
Purkinje cell in vitro source data 1.
- https://cdn.elifesciences.org/articles/63668/elife-63668-fig2-data1-v2.xlsx
Figure 2—figure supplement 1
In vitro cell-attached recordings of intrinsic activity in different Purkinje cell populations during development.
(A) Schematic of cell-attached recordings in vitro in lobules III and X (left) and example of a Purkinje cell filled with biocytin and stained afterwards (right). (B) Example traces of cell-attached …
Figure 2—figure supplement 2
Lobule III and ZebrinII– Purkinje cell simple spike rate results from a higher intrinsic and input driven component when compared with lobule X and ZebrinII+ Purkinje cells.
(A1) Purkinje cell intrinsic simple spike rate recorded in vitro for lobule III Purkinje cells (gray) and lobule X Purkinje cells (purple) and Purkinje cell simple spike firing rate recorded in vivo …
Figure 3 with 1 supplement
Purkinje cells located in the lobule III display a larger and elaborated dendritic tree.
(A) Photomicrographs of Purkinje cells filled with biocytin in lobule III and X (top panel); and sholl analysis of Purkinje cells located in lobules I/II/III (gray) or IX/X (purple) in six age …
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Figure 3—source data 1
Purkinje cell dendritic morphological source data 1.
- https://cdn.elifesciences.org/articles/63668/elife-63668-fig3-data1-v2.xlsx
Figure 3—figure supplement 1
Purkinje cell location in the cerebellum within each lobule.
(A) Photomicrographs of Purkinje cells filled with biocytin in lobule III; regions within lobule III (A = apex; B = base; S = sulcus), Scale bar = 500, 200 and 50 µm. (B1) Percentage of Purkinje cell…
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Figure 3—figure supplement 1—source data 1
Apex, Base, Sulcus source data 1.
- https://cdn.elifesciences.org/articles/63668/elife-63668-fig3-figsupp1-data1-v2.xlsx
Figure 4
Translocation of climbing fibers occurs earlier in the nodular regions of the cerebellum.
(A) Photomicrographs of lobule III and lobule X regions depicting climbing fiber (VGluT2 staining, cyan) and Purkinje cell (calbindin, red) development in five age groups. (B1) Molecular layer (ML) …
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Figure 4—source data 1
VGluT2 puncta source data 1.
- https://cdn.elifesciences.org/articles/63668/elife-63668-fig4-data1-v2.xlsx
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Figure 4—source data 2
Molecular layer thickness and climbing fiber height source data 2.
- https://cdn.elifesciences.org/articles/63668/elife-63668-fig4-data2-v2.xlsx
Figure 5 with 1 supplement
ZebrinII+ Purkinje cell axons are larger and less dense at all early postnatal ages.
(A) Photomicrographs of cerebellar sections demonstrating sparse labeling of Purkinje cells. Left panels are stained with antibodies against ZebrinII as well as red fluorescent protein (RFP) to …
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Figure 5—source data 1
Purkinje cell axonal morphological source data 1.
- https://cdn.elifesciences.org/articles/63668/elife-63668-fig5-data1-v2.xlsx
Figure 5—figure supplement 1
Random distribution of labeled Purkinje cells for axon analysis.
Sparsely labeled Purkinje cells are randomly distributed throughout the cerebellar cortex at P10, P14, and P21 as exemplified by three representative mice at each age. Purkinje cells are labeled …
Figure 6 with 4 supplements
Differential development of cerebellar regions impacts the maturation of cerebellar-specific behaviors.
(A1) Schematic illustration of eye movement recording setup. Mice are head-fixed in the center of a turntable (green arrow) for vestibular stimulation and surrounded by a random dotted pattern drum …
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Figure 6—source data 1
Eyeblink source data 1.
- https://cdn.elifesciences.org/articles/63668/elife-63668-fig6-data1-v2.xlsx
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Figure 6—source data 2
Eye movement source data 2.
- https://cdn.elifesciences.org/articles/63668/elife-63668-fig6-data2-v2.xlsx
Figure 6—figure supplement 1
Compensatory eye movements in young and adult mice.
Gain (top) and phase (bottom) of baseline performance of compensatory eye movements: (A1 and B1) the optokinetic reflex (OKR), (A2 and B2) the vestibulo-ocular reflex (VOR) and (A3 and B3) the …
Figure 6—figure supplement 2
Vestibulo-ocular reflex phase reversal gain, overnight consolidation, and gaze during training differ between juvenile and adult mice.
(A) Example trace of eye movement position in young (blue) and adult (green) mice before and after training in relation with the table position (black) at an amplitude of 5°. (B) Gain of 5 days of …
Figure 6—figure supplement 3
Polar plot for vestibulo-ocular reflex phase reversal gain.
Vestibulo-ocular reflex (VOR) phase reversal gain in (A) individual sessions and (B) averaged sessions per 5 days in young (blue) and adult (green) mice.
Figure 6—figure supplement 4
Eyeblink conditioning in young and adult mice.
(A1) Example traces of a single unconditioned response and (A2) a single conditioned response. Insets: mouse eye video captures show eyelid closure ranging from 0 (fully-open) to 1 (fully-closed). (B…
Figure 7
Summary of developmental timelines of Purkinje cell activity and morphology.
Left, selected features for physiological activity, represented by simple spike firing rate in vitro, simple spike rate in vivo and complex spike rate in vivo. Below the graphs periods of rapid …
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (Mus musculus) | Slc1a6-EGFP | Gong et al., 2003 | MMRRC: 012845-UCD | (Tg(Slc1a6-EGFP) HD185Gsat/Mmucd) |
| Strain, strain background (Mus musculus) | Pcp2-creERT2 | Institut Clinique de la Souris, France Jackson Laboratory | MGI:97508; ICS: 0273 | (Tg(Pcp2-creERT2)17.8.ICS) |
| Strain, strain background (Mus musculus) | Ai14 | The Jackson Laboratory | JAX: 007908 | (Gt(ROSA)26Sortm14(CAG-tdTomato)Hze/J) |
| Strain, strain background (Mus musculus) | C57BL/6 | The Jackson Laboratory Janvier Labs Charles River | JAX: 00055 Janvier: C57BL/6JRj CR: C57BL/6NCrl | |
| Antibody | Goat anti-ZebrinII/Aldolase C (Goat polyclonal) | Santa Cruz Biotechnology | Cat# sc-12065 RRID:AB_2242641 RRID:AB_2315622 | 1:1000 |
| Antibody | Mouse anti- Calbindin D-28K (Mouse monoclonal) | Swant | Cat# 300 RRID:AB_10000347 | 1:10,000 |
| Antibody | Guinea pig anti-VGluT2 (Guinea pig polyclonal) | Millipore | Cat# AB2251-l RRID:AB_2665454 | 1:2000 |
| Antibody | Rabbit anti-RFP (Rabbit polyclonal) | Rockland | Cat# 600-401-379 RRID:AB_2209751 | 1:1000 |
| Antibody | Cy3 Streptavidin | Jackson ImmunoResearch | Cat# 016-160-084 RRID:AB_233724 | 1:1000 |
| Antibody | Cy3-AffiniPure Donkey anti-Mouse (Mouse polyclonal) | Jackson ImmunoResearch | Cat# 715-165-150 RRID:AB_2340813 | 1:1000 |
| Antibody | Alexa Fluor 488-AffiniPure Donkey anti-Guinea Pig (Guinea pig polyclonal) | Jackson ImmunoResearch | Cat# 706-545-148 RRID:AB_2340472 | 1:1000 |
| Antibody | Donkey anti-goat Daylight 488 (Goat polyclonal) | Jackson ImmunoResearch | Cat# 705-486-147 RRID:AB_2616594 | 1:500 |
| Antibody | Cy3-AffiniPure Donkey Anti-Rabbit (Rabbit polyclonal) | Jackson ImmunoResearch | Cat# 711-165-152 RRID:AB_2307443 | 1:500 |
| Chemical compound, drug | 4’,6-Diamidine-2’-phenylindole dihydrochloride (DAPI) | Thermo Fisher Scientific | Cat# D3571 RRID:AB_2307455 | |
| Chemical compound, drug | Paraformaldehyde (PFA) | Millipore | Cat# 104005 | 4% |
| Chemical compound, drug | Biocytin | Sigma-Aldrich | Cat# B4261 | 1% |
| Chemical compound, drug | Evans Blue | Sigma-Aldrich | Cat# E2129 | 0.5% |
| Chemical compound, drug | Picrotoxin | Hello Bio Ltd | Cat# HB0506 | 100 µM |
| Chemical compound, drug | NBQX | Hello Bio Ltd | Cat# HB0442 | 10 µM |
| Chemical compound, drug | D-AP5 | Hello Bio Ltd | Cat# HB0225 | 50 µM |
| Chemical compound, drug | Mineral oil | Sigma-Aldrich | Cat# M3516 | |
| Chemical compound, drug | Tamoxifen | Sigma-Aldrich | Cat# T5648 | |
| Software, algorithm | FIJI (ImageJ) | National Institute of Health | RRID:SCR_002285 | |
| Software, algorithm | MATLAB 2008, 2016 | MathWorks | RRID:SCR_001622 | |
| Software, algorithm | ZEN digital Imaging for Light Microscopy | ZEISS | RRID:SCR_013672 | |
| Software, algorithm | Leica Application Suite X (LAS X) | Leica Microsystems | RRID:SCR_013673 | |
| Software, algorithm | Clampfit 10 | Molecular Devices | RRID:SCR_011323 | |
| Software, algorithm | Patchmaster | HEKA Electronics | RRID:SCR_000034 | |
| Software, algorithm | GraphPad Prism | GraphPad Software | RRID:SCR_002798 | |
| Software, algorithm | Compensatory eye movements analysis | Schonewille group | https://github.com/MSchonewille/iMove | |
| Software, algorithm | Eyeblink conditioning analysis | Neurasmus B.V. Rotterdam | RRID:SCR_021043 | |
| Other | Leica SM2000 R sliding microtome | Leica Biosystems | RRID:SCR_018456 | |
| Other | P-1000 Puller | Sutter Instrument | RRID:SCR_021042 | |
| Other | LSM 700 laser scanning confocal | ZEISS | RRID:SCR_017377 | |
| Other | SP5 confocal | Leica Microsystems | RRID:SCR_020233 | |
| Other | SP8 confocal | Leica Microsystems | RRID:SCR_018169 | |
| Other | Axio Imager.M2 | ZEISS | RRID:SCR_018876 |
Additional files
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Supplementary file 1
Data for in vivo, in vitro electrophysiology, anatomical and behavioral quantifications.
Descriptive values are represented as mean ± SEM. p=postnatal, Z−=ZebrinII negative, Z+=ZebrinII positive, LIII = lobule III, LX = lobule X, LI-III = lobules I/II/III, LIX-X = lobules IX/X.
- https://cdn.elifesciences.org/articles/63668/elife-63668-supp1-v2.docx
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Transparent reporting form
- https://cdn.elifesciences.org/articles/63668/elife-63668-transrepform-v2.docx
