Defective synaptic transmission causes disease signs in a mouse model of juvenile neuronal ceroid lipofuscinosis
- Jena University Hospital, Germany
- University Hospital Würzburg, Germany
- University of Münster, Germany
- University Hospital Frankfurt, Germany
- Sanford Research, United States
Figures
Figure 1 with 1 supplement
Anxiety-related behavior in Cln3Δex1-6 mice.
(A) Escape time to cross the rim barrier of a petri dish was increased in Cln3Δex1-6 mice compared to wt littermates indicating reduced explorative behavior (wt: n = 15/24 [7 month/14 month]; Cln3Δex…
Figure 1—figure supplement 1
Open field (OF) and elevated plus maze testing at the age of 7 months.
(A) Center visits and total movement of Cln3Δex1-6 mice during free exploration in the OF maze were unchanged in comparison to wt littermates at the age of 7 months (n = 9 vs 8; OF visits and OF …
Figure 2 with 2 supplements
Defective synaptic transmission in the amygdala of 14 months old Cln3Δex1-6mice.
(A) Schematic illustration of the recording situation in the lateral amygdala (LA). Whole-cell patch recordings were performed from principal neurons (PN) in the LA (rec) by local stimulation of …
Figure 2—figure supplement 1
In-out curve showing reduced eIPSC in the amygdala of Cln3Δex1-6 mice at the age of 14 months.
The amplitude of eIPSCs in Cln3Δex1-6 mice is reduced at all stimulation intensities that were tested (wt: n = 25/7 (number of recordings/number of mice); Cln3Δex1-6: n = 28/7; Student`s t-test). …
Figure 2—figure supplement 2
In-out curve showing reduced eEPSC in the amygdala of 14-month-old Cln3Δex1-6 mice.
eEPSC amplitude is reduced in Cln3Δex1-6 mice at all stimulation intensities that were tested (23/6 vs. 24/7; Student`s t-test. Exact values of N, dispersion and precision measures are provided in Su…
Figure 3 with 1 supplement
Loss of GABAergic interneurons in the amygdala of Cln3Δex1-6 mice at the age of 14 months.
In coronar brain slices the area of the amygdala core regions was morphologically identified by their characteristic shape and distribution of neuronal nuclei (dashed lines). Interneuron subtypes …
Figure 3—figure supplement 1
Density of NeuN-positive cells is unchanged in the amygdala of 14-month-old Cln3Δex1-6 mice (n = 9 vs. 8).
Exact p values as well as exact dispersion and precision measures are provided in Supplementary file 2.
Figure 4 with 1 supplement
Learning deficits and disturbed hippocampal synaptic transmission in Cln3Δex1-6 mice at the age of 14 months.
(A) For induction of fear conditioned and contextual learning a conditioned acoustic stimulus (tone) was paired with an unconditioned stimulus (electric shock) on day 0 in a specific environment …
Figure 4—figure supplement 1
Reduced eEPSC amplitudes in the hippocampus of 14 months old Cln3Δex1-6 mice.
The amplitude of eEPSCs in Cln3Δex1-6 mice is reduced at stimulation intensities ≥ 75 pA but is unchanged at stimulation intensities in the range of minimal stimulation (wt: n = 12/7; Cln3Δex1-6: n =…
Figure 5 with 2 supplements
Loss of GABAergic interneurons in the hippocampus of Cln3Δex1-6 mice at the age of 14 months.
Quantitative stereological analysis of interneuron subclasses revealed a reduced number of parvalbumin, neuropeptide Y and GAD-positive interneurons, whereas other markers were unchanged. Note that …
Figure 5—figure supplement 1
Density of interneurons within the hippocampal subfields of Cln3Δex1-6 mice and wt littermates.
Analysis of interneuron density within hippocampal subfields revealed a reduction of GAD in the CA1 region of Cln3Δex1-6 mice (n = 17 vs. 14; Student`s t-test) and a reduction of parvalbumin (n = 17 …
Figure 5—figure supplement 2
Density of NeuN-positive cells is unchanged in the hippocampus of 14-month-old Cln3Δex1-6 mice.
NeuN-positive neurons were analyzed in the principal cell layers of the hippocampus. Stereological analysis revealed unchanged count of NeuN-positive cells in the dentate gyrus and in the CA1 and …
Figure 6 with 2 supplements
Progressive ataxia and motor impairment in Cln3Δex1-6 mice.
(A, B) Mice were trained to climb a vertically stretched cord of 1 m length to reach a platform (climbing test). Mice used their limbs in an alternating manner that allows fast movement and rapid …
Figure 6—figure supplement 1
Grip strength of Cln3Δex1-6 mice and wt littermates at different ages.
Grip strength of the forelimbs is not affected in Cln3Δex1-6 mice. Grip strength measures of wt and Cln3Δex1-6 mice were unchanged between genotypes at all time points (3 months: n = 11 vs. 12; 7 …
Figure 6—figure supplement 2
Gait analysis of Cln3Δex1-6 mice and wt littermates.
Gait analysis of Cln3Δex1-6 mice at the age of 7 and 14 months revealed no differences as compared to wt mice. Right and left stride length (rsl and lsl), distance between hind feet (dbh) and angle …
Figure 7
Pathological firing of cerebellar Purkinje cells (PC) of Cln3Δex1-6 mice at the age of 14 months.
(A) Schematic drawing of neurons and connections in the cerebellar cortex. Spontaneous and evoked action potentials of PC were recorded in loose patch configuration. Granule cells (GC) give rise to …
Figure 8
Reduction of GAD-positive interneurons in the cerebellar molecular layer of 14 months old Cln3Δex1-6 mice.
Quantitative stereological analysis of interneuron subclasses and NeuN-positive neurons in coronar cerebellar slices revealed a reduced number of GAD-positive interneurons within the molecular layer …
Figure 9 with 1 supplement
Spinal networks are unaffected in Cln3Δex1-6 mice.
(A) Example traces for H-Reflex recordings in mice; the first major deflection is the motor response (M-wave) upon anterograde sciatic nerve stimulation, the second is the H-Reflex after …
Figure 9—figure supplement 1
Number of spinal cord interneurons are unaffected.
Quantitative analysis of parvalbumin- and calbindin-positive interneurons showed no difference of cell counts in the ventral (VH) or dorsal horn (DH) of the spinal cord in of wt and Cln3Δex1-6 mice …
Videos
Video 1
Rope climbing performance of a wt littermate and a Cln3Δex1-6 mouse.
The wt mouse notices and grasps the vertically spanned rope quickly and climbs up the rope using all four limbs in an alternating manner. The Cln3Δex1-6 mouse notices and grasps the rope but is …
Tables
Key resources table
| Reagent type | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (Mus musculus) | Cln3Δex1-6 mice | Mitchison et al., 1999. Neurobiol Dis, 10.1006/nbdi.1999.0267 | MGI:1933976 | Initial breeding pairs from: David A. Pearce Sanford Children's Health Research Center, Sanford Research, 2301 E 60th Street North, Sioux Falls, SD 57104, USA |
| Antibody | Anti-Calbindin | Swant, Marly, Switzerland; CB38 | RRID: AB_10000340 | Dilution: 1:5000, overnight at 4°C |
| Antibody | Anti-Calretinin | Swant, Marly, Switzerland; 7697 | RRID: AB_2619710 | Dilution: 1:5000, overnight at 4°C |
| Antibody | Anti-GAD65/67 | Merck Millipore, Darmstadt Germany; AB1511 | RRID: AB_90715 | Dilution: 1:8000, overnight at 4°C |
| Antibody | Anti-Neuropeptid Y (NPY) | Thermo Fisher Scientific, Waltham, USA; PA5-19568 | RRID: AB_10987237 | Dilution: 1:1000, overnight at 4°C |
| Antibody | Anti-Parvalbumin | Swant, Marly, Switzerland; PV27 | RRID: AB_2631173 | Dilution: 1:5000, overnight at 4°C |
| Antibody | Anti-Somatostatin | Acris Antibodies, Herford, Germany; AP01098SU-N | RRID: AB_1623011 | Dilution: 1:50, overnight at 4°C |
Additional files
-
Supplementary file 1
Animal groups and counts.
List of all animal groups used in the study including animal number and the experiments in which they were included. Note that not all animals in the list were finally included in the analysis (e.g. mice were allocated to different tests, exclusion due to unstable recordings or insufficient quality criteria for neurophysiology etc.). The exact number of mice used in the respective experiment is given in the figure legends. Cln3Δex1-6 mice and age-matched littermates where allocated in a randomized way to the respective groups before starting the experiments by a blinded member of the laboratory.
- https://doi.org/10.7554/eLife.28685.022
-
Supplementary file 2
Statistic report table.
Table containing information regarding distribution, statistical analysis method, exact p values, n-values and number of biological replicates, mean, SD, SEM, median and confidence intervals, and Cohen’s d effect size for all experiments.
- https://doi.org/10.7554/eLife.28685.023
-
Transparent reporting form
- https://doi.org/10.7554/eLife.28685.024
