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Results: 5

1.
Figure 3

Figure 3. Long-term depression but not long-term potentiation is independent of protein synthesis in Cyfip1 heterozygotes.. From: Haploinsufficiency of Cyfip1 Produces Fragile X-Like Phenotypes in Mice.

(A, B) LTD induced by PP-LFS in wild type (A) or Cyfip1 heterozygous (B) mice, in the absence (o) or presence (•) of the protein synthesis inhibitor cycloheximide (60 µM). The effect of cycloheximide is significantly different across genotypes. (C, D) LTP induced by HFS in wildtype (C) or Cyfip1 heterozygous (D) mice, in the absence (o) or presence (•) of cycloheximide. In all panels, the large arrow indicates onset of stimulation. (E, F) Rapamycin blocks LTP induced by HFS in wildtype (E) and Cyfip1 heterozygous (F) mice, as shown by incubating slices in the absence (o) or presence (•) of the mTOR inhibitor rapamycin (20 nM). Onset of stimulation is indicated by an arrow.

Ozlem Bozdagi, et al. PLoS One. 2012;7(8):e42422.
2.
Figure 2

Figure 2. Basal synaptic properties and long-term potentiation are normal but long-term depression is enhanced in Cyfip1 heterozygotes.. From: Haploinsufficiency of Cyfip1 Produces Fragile X-Like Phenotypes in Mice.

(A) Hippocampal slices from 4–6 weeks old wildtype (WT) or Cyfip1 heterozygous (Het) mice were analyzed for baseline synaptic properties, determined by input/output function, representing the relationship between stimulus intensity and the size of the field EPSP slope. (B) Paired-pulse facilitation in the Schaffer collateral-commissural pathway is not different between genotypes over the test interpulse interval of 50 ms. (C) HFS-induced LTP was not significantly different between wildtype (WT) or Cyfip1 heterozygous (Het) mice. (D) PP-LFS-induced LTD in Cyfip1 heterozygous mice was significantly increased. Inset: Representative EPSP traces recorded before stimulation (arrow) or 60 min after stimulation in wildtype and heterozygous animals (scale: 10 ms and 0.5 mV).

Ozlem Bozdagi, et al. PLoS One. 2012;7(8):e42422.
3.
Figure 5

Figure 5. Normal learning and memory in Morris Water Maze and in fear conditioning but enhanced extinction of inhibitory avoidance in Cyfip1 heterozygous mice.. From: Haploinsufficiency of Cyfip1 Produces Fragile X-Like Phenotypes in Mice.

(A) Mice were tested using the Morris Water Maze. Time (s) to travel to the target platform was not significantly different between genotypes. (B) Mice were tested for fear conditioning, with mice receiving shocks at 120 and 180 seconds during training. Testing was performed 24 hours later, in the same test chamber, without footshock. (C) Inhibitory avoidance was measured by latency to enter the dark side of the box associated with prior shock. Extinction of inhibitory avoidance is enhanced in the heterozygotes. The lower panel shows the experimental design. WT, wildtype mice; Het, heterozygous mice; acq, acquisition; ext, extinction; IA, inhibitory avoidance. *, P = 0.027.

Ozlem Bozdagi, et al. PLoS One. 2012;7(8):e42422.
4.
Figure 4

Figure 4. DHPG-induced long-term depression is not dependent on protein synthesis or mammalian Target of Rapamycin in Cyfip1 heterozygotes.. From: Haploinsufficiency of Cyfip1 Produces Fragile X-Like Phenotypes in Mice.

(A) LTD was induced by DHPG (50 µM for 5 minutes, indicated by the short horizontal bar) in hippocampal slices from wildtype (Wt) and Cyfip1 heterozygous (Het) mice. LTD is significantly enhanced in the heterozygotes as compared to wildtype. Inset: Representative EPSP traces recorded before (arrow) or 40 min after DHPG in wildtype and heterozygous animals (scale: 10 ms and 0.5 mV). (B, C) LTD was induced with DHPG in wildtype (B) or Cyfip1 heterozygous (C) mice, in the absence (o) or presence (•) of cycloheximide (Cyclohex, 60 µM, indicated by the long horizontal bar). Cycloheximide significantly inhibited LTD in slices from wildtype but not heterozygous animals. (D, E) LTD was induced by DHPG (50 µM, indicated by the short horizontal bar) in hippocampal slices from wildtype (D) or Cyfip1 heterozygous (E) mice, in the absence (o) or presence (•) of rapamycin (20 nM, indicated by the long horizontal bar). (F) LTD was induced by DHPG (50 µM, indicated by the short horizontal bar) in hippocampal slices from wildtype (o) or Cyfip1 heterozygous (•) mice, the latter in the absence (•) or presence (▪) of both MPEP (10 µM) and LY367385 (indicated by the long horizontal bar). Application of the mGluR1 and mGluR5 antagonists decreased the magnitude of DHPG-induced-LTD in heterozygotes.

Ozlem Bozdagi, et al. PLoS One. 2012;7(8):e42422.
5.
Figure 1

Figure 1. Generation and characterization of a mouse with disruption of the Cyfip1 gene.. From: Haploinsufficiency of Cyfip1 Produces Fragile X-Like Phenotypes in Mice.

(A) The genomic structure of Cyfip1 is shown to scale with larger horizontal boxes representing exons, and the first (ATG) and last (Stop) coding exons indicated. The diagram shows the site of the gene-trap insertion (identified as an LTR-flanked Trapping casette) in intron 1 (5′ to the first coding exon), in order to generate mice with a disruption of the Cyfip1 gene. (B) Synaptoneurosome preparations from wildtype (Wt) and Cyfip1 heterozygous (Het) mice were subjected to quantitative immunoblotting with an antibody to Cyfip1, with actin as a reference protein. The migration of molecular weight markers is shown on the left (in kDa). (C) Brain mRNA from wildtype (black bars) and Cyfip1 heterozygous (white bars) mice were subjected to qPCR for the indicated genes. (D) Quantification of Cyfip1 and Fmrp by immunoblotting of extracts from wildtype (black bars) and heterozygous (white bars) mice. *, P<0.05; **, P = 0.004.

Ozlem Bozdagi, et al. PLoS One. 2012;7(8):e42422.

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