RNG105/caprin1, an RNA granule protein for dendritic mRNA localization, is essential for long-term memory formation
- National Institute for Basic Biology, Japan
- SOKENDAI, Japan
- Okazaki Institute for Integrative Bioscience, Japan
- Tokyo University of Science, Japan
- Tokyo University of Pharmacy and Life Sciences, Japan
- Niigata University, Japan
- Kobe City College of Nursing, Japan
- Brain Science Institute, Japan
Figures
Figure 1 with 1 supplement
Generation of RNG105 conditional deletion mice.
(A) Gene structure of the Rng105 genome, targeting vector, floxed Rng105, and after excision of the floxed sequence. Arrowheads indicate PCR primers for genotyping. (B) PCR analysis of the indicated …
Figure 1—figure supplement 1
Reduced expression of RNG105 without the expression of truncated forms in the cerebrum of Camk2a-Cre;Rng105f/f mice.
(A) Reduction of Rng105 transcripts in Camk2a-Cre;Rng105f/f mice compared to Rng105f/f mice. RNA-seq read coverage across the Rng105 genome for triplicate RNA samples from the hippocampus of Rng105f/…
Figure 2
RNG105-deficient mice have small dendritic spines.
(A) Density of CA1 hippocampal neurons measured by nuclear staining in the SP. n = 11. Student's t-test, p=0.674. (B) CA1 pyramidal neurons imaged with Thy1-GFP. Scale bar, 50 µm. (C and D) Length …
Figure 3
RNG105 deficiency impairs structural plasticity of dendritic spines.
(A) Fluorescence images of cultured hippocampal neurons from Rng105f/f mice transfected with mCherry (left) or both mCherry and CMV-Cre (right), and immunostained for RNG105. Arrows indicate …
Figure 4
RNG105 deficiency reduces fEPSP amplitude in hippocampal CA1.
(A) Representative fEPSP traces evoked by different stimulation intensities. (B) Input-output (I/O) relationship between CA3 stimulation magnitude and CA1 fEPSP responses. n = 10 (Rng105f/f) and 13 …
Figure 5 with 1 supplement
Impaired habituation to a novel place and normal motor skill learning in Camk2a-Cre;Rng105f/f mice.
(A) Open field test. The number of line crossings during a 5 min trial on 3 consecutive days. n = 14 (Rng105f/f), 9 (Camk2a-Cre;Rng105f/f) and 10 (Camk2a-Cre;Rng105f/+). ****p<0.001 (genotype …
Figure 5—figure supplement 1
Normal anxiety-like behavior and reduced novel object preference in Camk2a-Cre;Rng105f/f mice.
(A) Light/dark transition test. Left, distance traveled in the light (L) and dark (D) chambers; middle, time spent in the light chamber; right, the number of transition during a 5 min trial. n = 13 …
Figure 6 with 1 supplement
RNG105 conditional deletion mice display impaired long-term memory.
(A−D) Morris water maze. Escape latency to the platform in visible (A) and hidden (B) platform tasks. Density plot of swim paths averaged from all mice (C) and the time spent in each quadrant (D) …
Figure 6—figure supplement 1
Camk2a-Cre;Rng105f/f mice exhibit less freezing behavior in a contextual fear conditioning test.
Freezing time in a test chamber, in which the mouse had received foot shock 5 days before. n = 8 (Rng105f/f) and 4 (Camk2a-Cre;Rng105f/f). ****p<0.001 (genotype effect) using two-way repeated …
Figure 7
Identification of somatically and dendritically enriched mRNAs (S- and D-mRNAs) in the hippocampal CA1.
(A) Schematic diagram of a mouse hippocampal slice. (B) A hippocampal slice from an adult mouse (P12 weeks) before (left) and after (right) isolation of CA1 SR. Nuclei are stained with Yo-Pro1 …
Figure 8
Reduced dendritic localization of D-mRNAs in RNG105-deficient mice.
(A) MA plots of mRNAs in the hippocampal CA1 of Rng105f/f mice (left) and Camk2a-Cre;Rng105f/f mice (right). (B) Heat maps showing DAI of each mRNA in Rng105f/f mice (left lane), in Camk2a-Cre;Rng105…
Figure 9
Dendritic accumulation of mRNAs classified in ‘regulation of Arf protein signal transduction’ and ‘structural constituent of ribosome’ is reduced by RNG105 deficiency.
(A and B) MA plots of mRNAs classified in ‘regulation of Arf protein signal transduction’ (A) and ‘structural constituent of ribosome’ (B) in Rng105f/f mice (left) and Camk2a-Cre;Rng105f/f mice …
Figure 10 with 1 supplement
RNG105 deficiency impairs AMPAR scaling in response to activity deprivation.
(A and B) Immunostaining for GluR1 (A) and GluR2 (B) in cultured neurons (9 DIV) from the cerebral cortex of E17.5 Rng105+/+ and Rng105−/− littermates. The neurons were cultured with (+) or without …
Figure 10—figure supplement 1
RNG105 deficiency impairs AMPAR scaling in response to activity deprivation (biotinylation assay).
(A) Total cell lysates of surface-biotinylated primary cultured neurons (9 DIV) from E17.5 wild-type mouse cerebral cortex (total), and avidin agarose beads-bound fractions of the lysates (surface), …
Videos
Video 1
RNG105 conditional deletion mice exhibit seizures.
This movie shows a seizure in a Camk2a-Cre;Rng105f/f mouse just after the contextual fear conditioning test (passive avoidance).
Additional files
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Supplementary file 1
Differential mRNA enrichment analysis between SP and SR of hippocampus − All mRNAs (36,701)
- https://doi.org/10.7554/eLife.29677.018
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Supplementary file 2
(A) mRNAs eliminated from the candidates for dendritic mRNAs. (B) mRNAs eliminated from the candidates for somatic mRNAs
- https://doi.org/10.7554/eLife.29677.019
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Supplementary file 3
(A) Dendritically enriched mRNAs (1,122). (B) Somatically enriched mRNAs (2,106). (C) mRNAs not significantly enriched in dendrites nor soma (2,814)
- https://doi.org/10.7554/eLife.29677.020
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Supplementary file 4
(A) Dendritic mRNAs classified in ‘regulation of Arf protein signal transduction’. (B) Dendritic mRNAs classified in ‘structural constituent of ribosome’. (C) Dendritic mRNAs classified in ‘translation’. (D) Dendritic mRNAs classified in ‘Fc gamma R-mediated phagocytosis’. (E) Dendritic mRNAs classified in ‘pleckstrin homology’. (F) Dendritic mRNAs classified in ‘chemotaxis’. (G) Dendritic mRNAs classified in ‘GTPase regulator activity’. (H) Dendritic mRNAs classified in ‘leukocyte activation’. (I) Dendritic mRNAs classified in ‘SH3’
- https://doi.org/10.7554/eLife.29677.021
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Supplementary file 5
(A) mRNAs for AMPA receptor subunits and dendritic mRNAs for AMPA receptor regulators. (B) SR-enriched mRNAs involved in membrane potential regulation
- https://doi.org/10.7554/eLife.29677.022
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Supplementary file 6
mRNAs reduced in Camk2a-Cre;Rng105f/f soma
- https://doi.org/10.7554/eLife.29677.023
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Supplementary file 7
Statistical reporting table
- https://doi.org/10.7554/eLife.29677.024
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Transparent reporting form
- https://doi.org/10.7554/eLife.29677.025
