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Rhes influences striatal cAMP/PKA-dependent signaling and synaptic plasticity in a gender-sensitive fashion.

Ghiglieri V, Napolitano F, Pelosi B, Schepisi C, Migliarini S, Di Maio A, Pendolino V, Mancini M, Sciamanna G, Vitucci D, Maddaloni G, Giampà C, Errico F, Nisticò R, Pasqualetti M, Picconi B, Usiello A - Sci Rep (2015)

Bottom Line: Corticostriatal LTP defects are exclusively found in A2AR/D2R-expressing MSNs of KO females, compared to KO males, an effect that is abolished by PKA inhibitors but not by the removal of circulating estrogens.This suggests that the synaptic alterations found in KO females could be triggered by an aberrant A2AR/cAMP/PKA activity, but not due to estrogen-mediated effect.Thus Rhes, a thyroid hormone-target gene, plays a relevant role in gender-specific synaptic and behavioral responses.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Philosophy, Human, Social, and Educational Sciences, University of Perugia, Perugia, Italy [2] Fondazione Santa Lucia IRCCS, Rome, Italy.

ABSTRACT
Mechanisms of gender-specific synaptic plasticity in the striatum, a brain region that controls motor, cognitive and psychiatric functions, remain unclear. Here we report that Rhes, a GTPase enriched in medium spiny neurons (MSNs) of striatum, alters the striatal cAMP/PKA signaling cascade in a gender-specific manner. While Rhes knockout (KO) male mice, compared to wild-type (WT) mice, had a significant basal increase of cAMP/PKA signaling pathway, the Rhes KO females exhibited a much stronger response of this pathway, selectively under the conditions of dopamine/adenosine-related drug challenge. Corticostriatal LTP defects are exclusively found in A2AR/D2R-expressing MSNs of KO females, compared to KO males, an effect that is abolished by PKA inhibitors but not by the removal of circulating estrogens. This suggests that the synaptic alterations found in KO females could be triggered by an aberrant A2AR/cAMP/PKA activity, but not due to estrogen-mediated effect. Consistent with increased cAMP signaling, D1R-mediated motor stimulation, haloperidol-induced catalepsy and caffeine-evoked hyper-activity are robustly enhanced in Rhes KO females compared to mutant males. Thus Rhes, a thyroid hormone-target gene, plays a relevant role in gender-specific synaptic and behavioral responses.

No MeSH data available.


Related in: MedlinePlus

Long-term potentiation in MSNs of Rhes mutant mice and morphological characterization.(A) Time-course plots of EPSP amplitude (left) and pairs of traces (right) demonstrate the presence of HFS-induced LTP in MSNs recorded from slices of male WT (n = 5) and KO (n = 7) mice (Student’s t-test, pre- vs. 30 min post-HFS, **p < 0.01). (B) Time course graphs of the EPSC amplitude (left) and pairs of traces (right) show the induction (n = 12) or the complete loss (n = 15) of LTP in MSNs from female KO mice compared with age-matched WT animals (n = 5) (Student’s t-test at 30 min post-HFS, WT vs. KO with LTP, p > 0.05, WT vs. KO with no LTP, ***p < 0.001). (C) In KO females application of a low-frequency stimulation (LFS) induced a decrease of EPSP amplitude in 5 of 9 MSNs recorded and a mild potentiation in the remaining 4 MSNs, whereas the same protocol induced no change of synaptic activity in all the MSNs (n = 7) recorded from WT females (Bonferroni post-hoc: WT vs. KO with LTP, *p < 0.05; **p < 0.01; ***p < 0.001; Student’s t-test at 30 min post-HFS: WT vs. KO with a depression of EPSP, ***p < 0.001). (D–F) Confocal laser scanning microscopy (CLSM) images of double-labeled immunofluorescence for biocytin and adenosine receptor (A2AR). Biocytin immunolabeling is visualized in red streptavidin-Cy3 immunofluorescence (D) and A2AR is visualized in green Cy2 immunofluorescence (E). The colocalization of A2AR and biocytin is showed in merged panel (F) as yellow fluorescence. (G) HFS protocol applied in the MSNs of ovariectomized (OVX) KO females induced LTP in 6 of 12 MSNs recorded (Student’s t-test at 30 min post-HFS: WT vs. KO OVX with no LTP, *p < 0.05). (H,I) Striatal ERα and ERβ, determined by Western blotting, in WT and KO male (n = 6 /genotype per ERα; n = 6 WT, 5 KO per EPβ) (H) and female (n = 5 WT, 6 KO per ERα; n = 6 /genotype per ERβ) (I) mice. The top panels show representative blots comparing the different genotypes. All data are expressed as mean ± SEM. Genotypes are as indicated.
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f4: Long-term potentiation in MSNs of Rhes mutant mice and morphological characterization.(A) Time-course plots of EPSP amplitude (left) and pairs of traces (right) demonstrate the presence of HFS-induced LTP in MSNs recorded from slices of male WT (n = 5) and KO (n = 7) mice (Student’s t-test, pre- vs. 30 min post-HFS, **p < 0.01). (B) Time course graphs of the EPSC amplitude (left) and pairs of traces (right) show the induction (n = 12) or the complete loss (n = 15) of LTP in MSNs from female KO mice compared with age-matched WT animals (n = 5) (Student’s t-test at 30 min post-HFS, WT vs. KO with LTP, p > 0.05, WT vs. KO with no LTP, ***p < 0.001). (C) In KO females application of a low-frequency stimulation (LFS) induced a decrease of EPSP amplitude in 5 of 9 MSNs recorded and a mild potentiation in the remaining 4 MSNs, whereas the same protocol induced no change of synaptic activity in all the MSNs (n = 7) recorded from WT females (Bonferroni post-hoc: WT vs. KO with LTP, *p < 0.05; **p < 0.01; ***p < 0.001; Student’s t-test at 30 min post-HFS: WT vs. KO with a depression of EPSP, ***p < 0.001). (D–F) Confocal laser scanning microscopy (CLSM) images of double-labeled immunofluorescence for biocytin and adenosine receptor (A2AR). Biocytin immunolabeling is visualized in red streptavidin-Cy3 immunofluorescence (D) and A2AR is visualized in green Cy2 immunofluorescence (E). The colocalization of A2AR and biocytin is showed in merged panel (F) as yellow fluorescence. (G) HFS protocol applied in the MSNs of ovariectomized (OVX) KO females induced LTP in 6 of 12 MSNs recorded (Student’s t-test at 30 min post-HFS: WT vs. KO OVX with no LTP, *p < 0.05). (H,I) Striatal ERα and ERβ, determined by Western blotting, in WT and KO male (n = 6 /genotype per ERα; n = 6 WT, 5 KO per EPβ) (H) and female (n = 5 WT, 6 KO per ERα; n = 6 /genotype per ERβ) (I) mice. The top panels show representative blots comparing the different genotypes. All data are expressed as mean ± SEM. Genotypes are as indicated.

Mentions: Then, we investigated the corticostriatal long-term potentiation (LTP) in male and female mutants. Delivery of HFS to corticostriatal fibers in Mg2+-free medium, a condition that favors NMDA receptor activation, was able to induce LTP with comparable amplitude and time course in WT and KO male mice (Fig. 4A; two-way ANOVA, genotype effect, F(1,187) = 0.42, p > 0.05). Conversely, LTP amplitude was significantly altered in a subpopulation of MSNs recorded from mutant females. Specifically, although HFS-induced LTP was similar to controls in 12 out of 27 neurons, a complete loss of potentiation was found in the remaining 15 MSNs recorded (Fig. 4B; two-way ANOVA, genotype effect, F(2,493) = 30.29, p < 0.001; time × genotype interaction, F(34,493) = 8.71, p < 0.001).


Rhes influences striatal cAMP/PKA-dependent signaling and synaptic plasticity in a gender-sensitive fashion.

Ghiglieri V, Napolitano F, Pelosi B, Schepisi C, Migliarini S, Di Maio A, Pendolino V, Mancini M, Sciamanna G, Vitucci D, Maddaloni G, Giampà C, Errico F, Nisticò R, Pasqualetti M, Picconi B, Usiello A - Sci Rep (2015)

Long-term potentiation in MSNs of Rhes mutant mice and morphological characterization.(A) Time-course plots of EPSP amplitude (left) and pairs of traces (right) demonstrate the presence of HFS-induced LTP in MSNs recorded from slices of male WT (n = 5) and KO (n = 7) mice (Student’s t-test, pre- vs. 30 min post-HFS, **p < 0.01). (B) Time course graphs of the EPSC amplitude (left) and pairs of traces (right) show the induction (n = 12) or the complete loss (n = 15) of LTP in MSNs from female KO mice compared with age-matched WT animals (n = 5) (Student’s t-test at 30 min post-HFS, WT vs. KO with LTP, p > 0.05, WT vs. KO with no LTP, ***p < 0.001). (C) In KO females application of a low-frequency stimulation (LFS) induced a decrease of EPSP amplitude in 5 of 9 MSNs recorded and a mild potentiation in the remaining 4 MSNs, whereas the same protocol induced no change of synaptic activity in all the MSNs (n = 7) recorded from WT females (Bonferroni post-hoc: WT vs. KO with LTP, *p < 0.05; **p < 0.01; ***p < 0.001; Student’s t-test at 30 min post-HFS: WT vs. KO with a depression of EPSP, ***p < 0.001). (D–F) Confocal laser scanning microscopy (CLSM) images of double-labeled immunofluorescence for biocytin and adenosine receptor (A2AR). Biocytin immunolabeling is visualized in red streptavidin-Cy3 immunofluorescence (D) and A2AR is visualized in green Cy2 immunofluorescence (E). The colocalization of A2AR and biocytin is showed in merged panel (F) as yellow fluorescence. (G) HFS protocol applied in the MSNs of ovariectomized (OVX) KO females induced LTP in 6 of 12 MSNs recorded (Student’s t-test at 30 min post-HFS: WT vs. KO OVX with no LTP, *p < 0.05). (H,I) Striatal ERα and ERβ, determined by Western blotting, in WT and KO male (n = 6 /genotype per ERα; n = 6 WT, 5 KO per EPβ) (H) and female (n = 5 WT, 6 KO per ERα; n = 6 /genotype per ERβ) (I) mice. The top panels show representative blots comparing the different genotypes. All data are expressed as mean ± SEM. Genotypes are as indicated.
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Related In: Results  -  Collection

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f4: Long-term potentiation in MSNs of Rhes mutant mice and morphological characterization.(A) Time-course plots of EPSP amplitude (left) and pairs of traces (right) demonstrate the presence of HFS-induced LTP in MSNs recorded from slices of male WT (n = 5) and KO (n = 7) mice (Student’s t-test, pre- vs. 30 min post-HFS, **p < 0.01). (B) Time course graphs of the EPSC amplitude (left) and pairs of traces (right) show the induction (n = 12) or the complete loss (n = 15) of LTP in MSNs from female KO mice compared with age-matched WT animals (n = 5) (Student’s t-test at 30 min post-HFS, WT vs. KO with LTP, p > 0.05, WT vs. KO with no LTP, ***p < 0.001). (C) In KO females application of a low-frequency stimulation (LFS) induced a decrease of EPSP amplitude in 5 of 9 MSNs recorded and a mild potentiation in the remaining 4 MSNs, whereas the same protocol induced no change of synaptic activity in all the MSNs (n = 7) recorded from WT females (Bonferroni post-hoc: WT vs. KO with LTP, *p < 0.05; **p < 0.01; ***p < 0.001; Student’s t-test at 30 min post-HFS: WT vs. KO with a depression of EPSP, ***p < 0.001). (D–F) Confocal laser scanning microscopy (CLSM) images of double-labeled immunofluorescence for biocytin and adenosine receptor (A2AR). Biocytin immunolabeling is visualized in red streptavidin-Cy3 immunofluorescence (D) and A2AR is visualized in green Cy2 immunofluorescence (E). The colocalization of A2AR and biocytin is showed in merged panel (F) as yellow fluorescence. (G) HFS protocol applied in the MSNs of ovariectomized (OVX) KO females induced LTP in 6 of 12 MSNs recorded (Student’s t-test at 30 min post-HFS: WT vs. KO OVX with no LTP, *p < 0.05). (H,I) Striatal ERα and ERβ, determined by Western blotting, in WT and KO male (n = 6 /genotype per ERα; n = 6 WT, 5 KO per EPβ) (H) and female (n = 5 WT, 6 KO per ERα; n = 6 /genotype per ERβ) (I) mice. The top panels show representative blots comparing the different genotypes. All data are expressed as mean ± SEM. Genotypes are as indicated.
Mentions: Then, we investigated the corticostriatal long-term potentiation (LTP) in male and female mutants. Delivery of HFS to corticostriatal fibers in Mg2+-free medium, a condition that favors NMDA receptor activation, was able to induce LTP with comparable amplitude and time course in WT and KO male mice (Fig. 4A; two-way ANOVA, genotype effect, F(1,187) = 0.42, p > 0.05). Conversely, LTP amplitude was significantly altered in a subpopulation of MSNs recorded from mutant females. Specifically, although HFS-induced LTP was similar to controls in 12 out of 27 neurons, a complete loss of potentiation was found in the remaining 15 MSNs recorded (Fig. 4B; two-way ANOVA, genotype effect, F(2,493) = 30.29, p < 0.001; time × genotype interaction, F(34,493) = 8.71, p < 0.001).

Bottom Line: Corticostriatal LTP defects are exclusively found in A2AR/D2R-expressing MSNs of KO females, compared to KO males, an effect that is abolished by PKA inhibitors but not by the removal of circulating estrogens.This suggests that the synaptic alterations found in KO females could be triggered by an aberrant A2AR/cAMP/PKA activity, but not due to estrogen-mediated effect.Thus Rhes, a thyroid hormone-target gene, plays a relevant role in gender-specific synaptic and behavioral responses.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Philosophy, Human, Social, and Educational Sciences, University of Perugia, Perugia, Italy [2] Fondazione Santa Lucia IRCCS, Rome, Italy.

ABSTRACT
Mechanisms of gender-specific synaptic plasticity in the striatum, a brain region that controls motor, cognitive and psychiatric functions, remain unclear. Here we report that Rhes, a GTPase enriched in medium spiny neurons (MSNs) of striatum, alters the striatal cAMP/PKA signaling cascade in a gender-specific manner. While Rhes knockout (KO) male mice, compared to wild-type (WT) mice, had a significant basal increase of cAMP/PKA signaling pathway, the Rhes KO females exhibited a much stronger response of this pathway, selectively under the conditions of dopamine/adenosine-related drug challenge. Corticostriatal LTP defects are exclusively found in A2AR/D2R-expressing MSNs of KO females, compared to KO males, an effect that is abolished by PKA inhibitors but not by the removal of circulating estrogens. This suggests that the synaptic alterations found in KO females could be triggered by an aberrant A2AR/cAMP/PKA activity, but not due to estrogen-mediated effect. Consistent with increased cAMP signaling, D1R-mediated motor stimulation, haloperidol-induced catalepsy and caffeine-evoked hyper-activity are robustly enhanced in Rhes KO females compared to mutant males. Thus Rhes, a thyroid hormone-target gene, plays a relevant role in gender-specific synaptic and behavioral responses.

No MeSH data available.


Related in: MedlinePlus