Journal: CNS Neuroscience & Therapeutics

Article Title: Cortical dopamine D5 receptors regulate neuronal circuit oscillatory activity and memory in rats

doi: 10.1111/cns.14210

Figure Lengend Snippet: Knockdown of the D5R in PFC increases GSK‐3β activity. (A) The experimental timeline is shown. (B) Electrode placements in the PFC, OFC, thalamus, and HIP. (C) Representative images and quantification of fluorescence showing reduced PFC expression of the D5R (top panels) and GSK‐3β phosphorylation at Ser9 (bottom panels) following D5R‐shRNA induced knockdown. N = 7 rats/group (D) Images showing drd5 gene expression in control (left panels) animals or following drd5 mRNA knockdown (right panels). (E) Quantification of the number of drd5 mRNA expressing cells. N = 3 rats/group, 2 slices/rat. Quantified data are expressed as percent control. ** p < 0.01, *** p < 0.001 Student's t test.

Article Snippet: Subsequently, adjacent slices were incubated in primary antibodies rabbit anti‐pGSK‐3β (Ser9) (catalogue #ab9107166, 1:200, Abcam) or rabbit anti‐D5R (catalogue #ADR‐005, 1:200, Alomone Labs) for 60 h at 4°C.

Techniques: Knockdown, Activity Assay, Fluorescence, Expressing, shRNA, Control

Journal: CNS Neuroscience & Therapeutics

Article Title: Cortical dopamine D5 receptors regulate neuronal circuit oscillatory activity and memory in rats

doi: 10.1111/cns.14210

Figure Lengend Snippet: Effect of PFC D5R knockdown on oscillatory power activity in rats. Effect of PFC D5R knockdown on oscillatory power activity in rats. (A–D) Power spectra (left and center panels) and quantification of power (right panels) are shown. PFC D5R knockdown had the following regional effects on spectral power, (A) increased theta power in PFC, (B) increased theta power in OFC, and (C) increased theta power in HIP. (D) No effects of PFC D5R knockdown were evident in spectral power in the thalamus. Power curves are presented as normalized data with jackknife estimates of SEM shown as shaded areas. Quantified data are expressed as percent control. N = 7–8 rats/group, 1–2 electrodes/region/rat. * p < 0.05 Student's t test.

Article Snippet: Subsequently, adjacent slices were incubated in primary antibodies rabbit anti‐pGSK‐3β (Ser9) (catalogue #ab9107166, 1:200, Abcam) or rabbit anti‐D5R (catalogue #ADR‐005, 1:200, Alomone Labs) for 60 h at 4°C.

Techniques: Knockdown, Activity Assay, Control

Journal: CNS Neuroscience & Therapeutics

Article Title: Cortical dopamine D5 receptors regulate neuronal circuit oscillatory activity and memory in rats

doi: 10.1111/cns.14210

Figure Lengend Snippet: Effect of PFC D5R knockdown on oscillatory coherence in rats. (A–F) Coherence spectra (left panels) and quantification (right panels) are shown. (A) PFC D5R knockdown increased PFC‐OFC theta coherence, and (B) reduced PFC–thalamus high gamma coherence. (C–F) No significant effects on coherence were observed between PFC–HIP, OFC‐HIP, OFC‐thalamus, or thalamus‐HIP. Coherence curves are presented with jackknife estimates of SEM shown as shaded areas. Quantified data are expressed as percent control. N = 7–8 rats/group, 1–2 electrodes/region/rat. * p < 0.05, ** p < 0.01 Student's t test.

Article Snippet: Subsequently, adjacent slices were incubated in primary antibodies rabbit anti‐pGSK‐3β (Ser9) (catalogue #ab9107166, 1:200, Abcam) or rabbit anti‐D5R (catalogue #ADR‐005, 1:200, Alomone Labs) for 60 h at 4°C.

Techniques: Knockdown, Control

Journal: CNS Neuroscience & Therapeutics

Article Title: Cortical dopamine D5 receptors regulate neuronal circuit oscillatory activity and memory in rats

doi: 10.1111/cns.14210

Figure Lengend Snippet: D5R knockdown in PFC induces deficits in learning and memory. (A) D5R knockdown in PFC induced deficits in object recognition memory in the NOR test. (B) Impaired spatial memory in the OL test was also evident. (C) D5R knockdown also induced impairment in associative recognition memory when tested in the OiP. (D) In the Y‐maze, D5R knockdown had no effects on short‐term recognition memory (left panel) but resulted in impairment on long‐term memory (right panel). N = 7–8 rats/group, * p < 0.05, ** p < 0.01, *** p < 0.001 Student's t test.

Article Snippet: Subsequently, adjacent slices were incubated in primary antibodies rabbit anti‐pGSK‐3β (Ser9) (catalogue #ab9107166, 1:200, Abcam) or rabbit anti‐D5R (catalogue #ADR‐005, 1:200, Alomone Labs) for 60 h at 4°C.

Techniques: Knockdown

Journal: CNS Neuroscience & Therapeutics

Article Title: Cortical dopamine D5 receptors regulate neuronal circuit oscillatory activity and memory in rats

doi: 10.1111/cns.14210

Figure Lengend Snippet: Knockdown of the D5R in PFC increases GSK‐3β activity. (A) The experimental timeline is shown. (B) Electrode placements in the PFC, OFC, thalamus, and HIP. (C) Representative images and quantification of fluorescence showing reduced PFC expression of the D5R (top panels) and GSK‐3β phosphorylation at Ser9 (bottom panels) following D5R‐shRNA induced knockdown. N = 7 rats/group (D) Images showing drd5 gene expression in control (left panels) animals or following drd5 mRNA knockdown (right panels). (E) Quantification of the number of drd5 mRNA expressing cells. N = 3 rats/group, 2 slices/rat. Quantified data are expressed as percent control. ** p < 0.01, *** p < 0.001 Student's t test.

Article Snippet: We hybridized the sections with the D5R mRNA probe (Cat #589931, Advanced Cell Diagnostics) for 2 h at 40°C then kept overnight in 5× SSC at room temperature.

Techniques: Activity Assay, Fluorescence, Expressing, shRNA

Journal: CNS Neuroscience & Therapeutics

Article Title: Cortical dopamine D5 receptors regulate neuronal circuit oscillatory activity and memory in rats

doi: 10.1111/cns.14210

Figure Lengend Snippet: Effect of PFC D5R knockdown on oscillatory power activity in rats. Effect of PFC D5R knockdown on oscillatory power activity in rats. (A–D) Power spectra (left and center panels) and quantification of power (right panels) are shown. PFC D5R knockdown had the following regional effects on spectral power, (A) increased theta power in PFC, (B) increased theta power in OFC, and (C) increased theta power in HIP. (D) No effects of PFC D5R knockdown were evident in spectral power in the thalamus. Power curves are presented as normalized data with jackknife estimates of SEM shown as shaded areas. Quantified data are expressed as percent control. N = 7–8 rats/group, 1–2 electrodes/region/rat. * p < 0.05 Student's t test.

Article Snippet: We hybridized the sections with the D5R mRNA probe (Cat #589931, Advanced Cell Diagnostics) for 2 h at 40°C then kept overnight in 5× SSC at room temperature.

Techniques: Activity Assay

Journal: CNS Neuroscience & Therapeutics

Article Title: Cortical dopamine D5 receptors regulate neuronal circuit oscillatory activity and memory in rats

doi: 10.1111/cns.14210

Figure Lengend Snippet: Effect of PFC D5R knockdown on oscillatory coherence in rats. (A–F) Coherence spectra (left panels) and quantification (right panels) are shown. (A) PFC D5R knockdown increased PFC‐OFC theta coherence, and (B) reduced PFC–thalamus high gamma coherence. (C–F) No significant effects on coherence were observed between PFC–HIP, OFC‐HIP, OFC‐thalamus, or thalamus‐HIP. Coherence curves are presented with jackknife estimates of SEM shown as shaded areas. Quantified data are expressed as percent control. N = 7–8 rats/group, 1–2 electrodes/region/rat. * p < 0.05, ** p < 0.01 Student's t test.

Article Snippet: We hybridized the sections with the D5R mRNA probe (Cat #589931, Advanced Cell Diagnostics) for 2 h at 40°C then kept overnight in 5× SSC at room temperature.

Techniques:

Journal: CNS Neuroscience & Therapeutics

Article Title: Cortical dopamine D5 receptors regulate neuronal circuit oscillatory activity and memory in rats

doi: 10.1111/cns.14210

Figure Lengend Snippet: D5R knockdown in PFC induces deficits in learning and memory. (A) D5R knockdown in PFC induced deficits in object recognition memory in the NOR test. (B) Impaired spatial memory in the OL test was also evident. (C) D5R knockdown also induced impairment in associative recognition memory when tested in the OiP. (D) In the Y‐maze, D5R knockdown had no effects on short‐term recognition memory (left panel) but resulted in impairment on long‐term memory (right panel). N = 7–8 rats/group, * p < 0.05, ** p < 0.01, *** p < 0.001 Student's t test.

Article Snippet: We hybridized the sections with the D5R mRNA probe (Cat #589931, Advanced Cell Diagnostics) for 2 h at 40°C then kept overnight in 5× SSC at room temperature.

Techniques:

Journal: Frontiers in Neuroanatomy

Article Title: D1- and D2-type dopamine receptors are immunolocalized in pial and layer I astrocytes in the rat cerebral cortex

doi: 10.3389/fnana.2023.1111008

Figure Lengend Snippet: Information on primary antibodies.

Article Snippet: Dopamine receptor D5 (D5R) , C-terminal domain (aa 455–472) of rat D5R , Santa Cruz biotechnology; goat polyclonal; Cat# (R-18) sc-1441, RRID:AB_673640 , 1:300.

Techniques: Purification

Journal: Frontiers in Neuroanatomy

Article Title: D1- and D2-type dopamine receptors are immunolocalized in pial and layer I astrocytes in the rat cerebral cortex

doi: 10.3389/fnana.2023.1111008

Figure Lengend Snippet: Distribution of each dopamine receptor in the motor cortex and caudate putamen. (A) Representative immunohistochemistry of the frontal cortex for D1R. High-magnification images of rectangles indicated with E and I are shown in (E,I) . Moderate to strong D1R-immunoreactivities were observed in the pial surface of the cerebral cortex, layer (VI), and caudate putamen. (B) Representative immunohistochemistry of the frontal cortex for D2R. High magnification images of rectangles indicated with F and J are shown in (F) and (J) . (C) Representative immunohistochemistry of the frontal cortex for D4R. High magnification images of rectangles indicated with G and K are shown in (G) and (K) . (D) Representative immunohistochemistry of the frontal cortex for D5R. High magnification images of rectangles indicated with H and L are shown in (H) and (L) . (E–H) Immunohistochemical images of the cortical surface for D1R (E) , D2R (F) , D4R (G) , and D5R (H) . (I) Immunohistochemical images of the cortical layer V for D1R. Neuronal somata show weak immunopositive signals for D1R (arrowheads). (J) Immunohistochemical images of the cortical layer V for D2R. Neuronal somata show weak immunoreactivity for D2R (arrowheads). (K,L) Immunohistochemical images of the cortical layer V for D4R or D5R. Many neuronal somata (arrowheads) and apical dendrites (arrows) show immunoreactivity for D4R or D5R. Scale bars: 500 μm (A–D) ; 125 μm (E–H) ; 50 μm (I–L) .

Article Snippet: Dopamine receptor D5 (D5R) , C-terminal domain (aa 455–472) of rat D5R , Santa Cruz biotechnology; goat polyclonal; Cat# (R-18) sc-1441, RRID:AB_673640 , 1:300.

Techniques: Immunohistochemistry, Immunohistochemical staining

Journal: Frontiers in Neuroanatomy

Article Title: D1- and D2-type dopamine receptors are immunolocalized in pial and layer I astrocytes in the rat cerebral cortex

doi: 10.3389/fnana.2023.1111008

Figure Lengend Snippet: D5R-immunoreactivity in astrocytes. (A,B) Moderate D5R-immunoreactivities of the cortical surface of the prelimbic (A) and somatosensory (B) areas. (C,D) Confocal imaging of double immunostaining for D5R (magenta) and GS (green) indicated D5R-immunoreactivities biased toward the parenchymal side within pial astrocytes (arrowheads) in the prelimbic area. Processes of pial astrocytes (arrows) had only a few D5R-immunoreactivities. (E,F) D5R-immunoreactivities in the parenchymal side within pial astrocytes (arrowheads) in the somatosensory area. Processes of pial astrocytes (arrows) had only a few D5R-immunoreactivities. (G,H) In layer I of the primary motor area, astrocytes showed immunoreactivities for D5R (arrowheads). Strong and granular immunoreactivities for D5R were observed (magenta), which were in close vicinity of GS-positive structure (green), but not overlapped. (I,J) Confocal imaging of double immunostaining for D5R (magenta) and GS (green) indicated D5R-immunoreactivities in layer I astrocytes (arrows) in the primary motor area. (K,L) In layer I of the primary motor area, GS-positive astrocytes showed weak immunoreactivities for D5R (arrows). (M,N) In layer V of the primary motor area, GS-positive protoplasmic astrocytes (arrows) showed weak D5R-immunoreactivities. (O,P) In layer V of the primary motor area, strong D5R-immunoreactivities were observed in pyramidal cell-like large cells (asterisks) and their processes (arrows). Nuclei were stained with Hoechst33342. Scale bars: 20 μm (A,B) ; 10 μm (C–F,K–P) ; 5 μm (G–J) .

Article Snippet: Dopamine receptor D5 (D5R) , C-terminal domain (aa 455–472) of rat D5R , Santa Cruz biotechnology; goat polyclonal; Cat# (R-18) sc-1441, RRID:AB_673640 , 1:300.

Techniques: Imaging, Double Immunostaining, Staining

Journal: Frontiers in Neuroanatomy

Article Title: D1- and D2-type dopamine receptors are immunolocalized in pial and layer I astrocytes in the rat cerebral cortex

doi: 10.3389/fnana.2023.1111008

Figure Lengend Snippet: Information on primary antibodies.

Article Snippet: The anti-D5R antibody is a goat polyclonal antibody against a C-terminal domain of the rat D5R, comprising amino acids 455–472 (Ricci et al., ) (1:300, Cat# (R-18) sc-1441, RRID:AB_673640 , Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Techniques: Purification

Journal: Frontiers in Neuroanatomy

Article Title: D1- and D2-type dopamine receptors are immunolocalized in pial and layer I astrocytes in the rat cerebral cortex

doi: 10.3389/fnana.2023.1111008

Figure Lengend Snippet: Distribution of each dopamine receptor in the motor cortex and caudate putamen. (A) Representative immunohistochemistry of the frontal cortex for D1R. High-magnification images of rectangles indicated with E and I are shown in (E,I) . Moderate to strong D1R-immunoreactivities were observed in the pial surface of the cerebral cortex, layer (VI), and caudate putamen. (B) Representative immunohistochemistry of the frontal cortex for D2R. High magnification images of rectangles indicated with F and J are shown in (F) and (J) . (C) Representative immunohistochemistry of the frontal cortex for D4R. High magnification images of rectangles indicated with G and K are shown in (G) and (K) . (D) Representative immunohistochemistry of the frontal cortex for D5R. High magnification images of rectangles indicated with H and L are shown in (H) and (L) . (E–H) Immunohistochemical images of the cortical surface for D1R (E) , D2R (F) , D4R (G) , and D5R (H) . (I) Immunohistochemical images of the cortical layer V for D1R. Neuronal somata show weak immunopositive signals for D1R (arrowheads). (J) Immunohistochemical images of the cortical layer V for D2R. Neuronal somata show weak immunoreactivity for D2R (arrowheads). (K,L) Immunohistochemical images of the cortical layer V for D4R or D5R. Many neuronal somata (arrowheads) and apical dendrites (arrows) show immunoreactivity for D4R or D5R. Scale bars: 500 μm (A–D) ; 125 μm (E–H) ; 50 μm (I–L) .

Article Snippet: The anti-D5R antibody is a goat polyclonal antibody against a C-terminal domain of the rat D5R, comprising amino acids 455–472 (Ricci et al., ) (1:300, Cat# (R-18) sc-1441, RRID:AB_673640 , Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Techniques: Immunohistochemistry, Immunohistochemical staining

Journal: Frontiers in Neuroanatomy

Article Title: D1- and D2-type dopamine receptors are immunolocalized in pial and layer I astrocytes in the rat cerebral cortex

doi: 10.3389/fnana.2023.1111008

Figure Lengend Snippet: D5R-immunoreactivity in astrocytes. (A,B) Moderate D5R-immunoreactivities of the cortical surface of the prelimbic (A) and somatosensory (B) areas. (C,D) Confocal imaging of double immunostaining for D5R (magenta) and GS (green) indicated D5R-immunoreactivities biased toward the parenchymal side within pial astrocytes (arrowheads) in the prelimbic area. Processes of pial astrocytes (arrows) had only a few D5R-immunoreactivities. (E,F) D5R-immunoreactivities in the parenchymal side within pial astrocytes (arrowheads) in the somatosensory area. Processes of pial astrocytes (arrows) had only a few D5R-immunoreactivities. (G,H) In layer I of the primary motor area, astrocytes showed immunoreactivities for D5R (arrowheads). Strong and granular immunoreactivities for D5R were observed (magenta), which were in close vicinity of GS-positive structure (green), but not overlapped. (I,J) Confocal imaging of double immunostaining for D5R (magenta) and GS (green) indicated D5R-immunoreactivities in layer I astrocytes (arrows) in the primary motor area. (K,L) In layer I of the primary motor area, GS-positive astrocytes showed weak immunoreactivities for D5R (arrows). (M,N) In layer V of the primary motor area, GS-positive protoplasmic astrocytes (arrows) showed weak D5R-immunoreactivities. (O,P) In layer V of the primary motor area, strong D5R-immunoreactivities were observed in pyramidal cell-like large cells (asterisks) and their processes (arrows). Nuclei were stained with Hoechst33342. Scale bars: 20 μm (A,B) ; 10 μm (C–F,K–P) ; 5 μm (G–J) .

Article Snippet: The anti-D5R antibody is a goat polyclonal antibody against a C-terminal domain of the rat D5R, comprising amino acids 455–472 (Ricci et al., ) (1:300, Cat# (R-18) sc-1441, RRID:AB_673640 , Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Techniques: Imaging, Double Immunostaining, Staining

Journal: Biomedicines

Article Title: Intrarenal Dopaminergic System Is Dysregulated in SS- Resp18 mutant Rats

doi: 10.3390/biomedicines11010111

Figure Lengend Snippet: SS- Resp18 mutant rats have decreased renal D1-like receptor protein expression: SS and SS- Resp18 mutant rats were maintained on a high-salt diet for six weeks, and then the rat kidneys were harvested. Kidney protein lysates were immunoblotted for ( A ) D1R and D5R protein in SS and SS- Resp18 mutant rats, and ( B , C ) respective expressions were quantified by densitometry ( n = 6). Data are mean ± SEM. p = 0.011, p = 0.0003, vs. SS- Resp18 mutant rats, t -test.

Article Snippet: The following primary antibodies were used: D1R (EMD Millipore, Burlington, MA, USA, #MAB5290), D5R (EMD Millipore, #MAB5292), and GAPDH (Cell Signaling Technology, Danvers, MA, USA, #14C10).

Techniques: Mutagenesis, Expressing