Journal: Journal for Immunotherapy of Cancer

Article Title: Pharmacologic targeting of the dopamine D2 receptor impacts the efficacy of immune checkpoint blockade in melanoma

doi: 10.1136/jitc-2025-014080

Figure Lengend Snippet: Graphical abstract. Figure created in BioRender. BRC, bromocriptine; D2R, D2 receptor; HAL, haloperidol; ICI, immune checkpoint inhibitor; IL, interleukin; MCP, metoclopramide; MHC, major histocompatibility complex; PRL, prolactin.

Article Snippet: Samples were loaded in 10% precast polyacrylamide gels (Bio-Rad, cat. #4561036), run at 100 V in Tris/Glycine/sodium dodecyl sulfate (SDS) buffer (Bio-Rad 1610732), transferred to polyvinylidene fluoride (PVDF) membrane (Bio-Rad, cat. #1704272), blocked for 1 hour with 5% dry milk in tris buffered saline-Tween (TBS-T), and probed for PRL (R&D, cat. #AF1445), D2R (Proteintech, cat. #55084-1-AP), and β-Actin-HRP (BioLegend, cat. #664803) in blocking buffer.

Techniques: Immunopeptidomics

Journal: Journal for Immunotherapy of Cancer

Article Title: Pharmacologic targeting of the dopamine D2 receptor impacts the efficacy of immune checkpoint blockade in melanoma

doi: 10.1136/jitc-2025-014080

Figure Lengend Snippet: Tumor immune microenvironment in prolactin-locus ICI non-responder and responder models. ( A ) Experimental scheme. B16F0-bearing (CC51xB6)F1 and B6 mice received 100 µg αCTLA-4 and 200 µg αPD-1 on days 3, 6, and 10 after tumor inoculation. On day 13 after tumor inoculation, tumors were enriched for CD45+ TILs then processed for scRNA-seq. ( B ) UMAP and ( C ) composition analysis showing the breakdown of B6U, B6T, CC51U, CC51T annotated cell types overlaying the UMAP ( B ) and proportions of each cell type cluster. For each experimental condition, cell proportions (Y axis) were calculated by dividing the number of cells of a given type by the total number of cells. B6U, B6 untreated; B6T, B6 ICI-treated; CC51U, (CC51xB6)F1 untreated; CC51T, (CC51xB6)F1 ICI-treated. Cell types annotated manually using canonical markers. ( D ) Expression levels of M1 and M2 markers in the MΦ cluster. ( E ) Volcano plot highlighting DEGs in the CD20− CD8+ T cluster between B6T and CC51T. Significant DEG (padj≤0.05, |log2FC | ≥ 1) were determined using FindMarkers and are colored (red=upregulated in B6T; purple=upregulated in CC51T). ( F ) Expression levels of Prlr, Drd2, Htr3a and Htr4 in all clusters. αCTLA-4, anti-cytotoxic T-lymphocyte-associated protein 4; αPD-1, anti-programmed cell death protein-1; B, B cells; B16, B16 F0 cells; CD4+ T, CD4+ T cells; CD20- CD8+ T, CD8+ T cells; CD20+ CD8+ T, CD20+ CD8 T cells; cDC1, conventional dendritic cells, type I; DEGs, differentially expressed genes; Drd2, dopamine D2 receptor; Htr3a, serotonin 5HT3 receptor; Htr4, 5HT4 receptors; ICI, immune checkpoint inhibitor; MΦ, macrophages; MDSC, myeloid-derived suppressor cells; Mono, monocytes; NK T, natural killer T cells; Prlr, prolactin receptor; scRNA-seq, single-cell RNA sequencing; TILs, tumor-infiltrating leukocytes.

Article Snippet: Samples were loaded in 10% precast polyacrylamide gels (Bio-Rad, cat. #4561036), run at 100 V in Tris/Glycine/sodium dodecyl sulfate (SDS) buffer (Bio-Rad 1610732), transferred to polyvinylidene fluoride (PVDF) membrane (Bio-Rad, cat. #1704272), blocked for 1 hour with 5% dry milk in tris buffered saline-Tween (TBS-T), and probed for PRL (R&D, cat. #AF1445), D2R (Proteintech, cat. #55084-1-AP), and β-Actin-HRP (BioLegend, cat. #664803) in blocking buffer.

Techniques: Expressing, Derivative Assay, Single Cell, RNA Sequencing

Journal: Journal of insect physiology

Article Title: Neurochemical regulation of Aedes aegypti salivary gland function.

doi: 10.1016/j.jinsphys.2021.104193

Figure Lengend Snippet: Fig. 5. Immunolocalization of DA receptor in the isolated A. aegypti salivary gland with rat D1- receptor polyclonal antibody. (A) overview of sali vary glands at low magnification with the labels for salivary duct (SD), lateral lobes (LL), proximal lateral lobes (PLL), distal lateral lobes (DLL), median lobe (ML). Square insert in panel A is a representative image (n = 5) of gland treated with secondary anti body only (negative control). Hashtags correspond to the location of the magnified image for the median lobe (B), dorsal lateral lobe (C), proximal lateral lobe (D). Positive staining (green) of the salivary gland with the human D1 antibody was observed in different patterns for all lobes. DAPI counterstaining (blue) was used to localize nuclei. Scale bars are specific for each panel and images are representative of 5 individual replicates. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: After washing, the glands were incubated with rat anti-D1 or anti-D2 dopamine receptor polyclonal antibody at a dilution factor of 1:50 (Alamone Labs, Jerusalem, Israel) for 24 h at 4◦.

Techniques: Isolation, Negative Control, Staining

Journal: Journal of insect physiology

Article Title: Neurochemical regulation of Aedes aegypti salivary gland function.

doi: 10.1016/j.jinsphys.2021.104193

Figure Lengend Snippet: Fig. 6. Immunolocalization of DA receptor in the isolated A. aegypti salivary gland with rat D2- receptor polyclonal antibody. (A) overview of sali vary glands at low magnification with the labels for salivary duct (SD), lateral lobes (LL), proximal lateral lobes (PLL), distal lateral lobes (DLL), median lobe (ML). Hashtags correspond to the location of the magnified image for the median lobe (B), dorsal lateral lobe (C), proximal lateral lobe (D). Positive staining (green) of the salivary gland with the human D2 antibody was observed in different patterns for all lobes. DAPI counterstaining (blue) was used to localize nuclei. Scale bars are specific for each panel and images are representative of 5 individual repli cates. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: After washing, the glands were incubated with rat anti-D1 or anti-D2 dopamine receptor polyclonal antibody at a dilution factor of 1:50 (Alamone Labs, Jerusalem, Israel) for 24 h at 4◦.

Techniques: Isolation, Staining

Journal: Neoplasia (New York, N.Y.)

Article Title: Role of Dopamine Receptors in the Anticancer Activity of ONC201.

doi: 10.1016/j.neo.2017.10.002

Figure Lengend Snippet: Figure 1. D2-like receptors are expressed and functional in colorectal cancer cells. (A) Assessment of DRD2 mRNA expression in different cancer tissue samples using cBioportal. (B) Viability assessment after 72 hours of treatment with dopamine or sumanirole, as indicated in legend. Western blot analyses of PKA substrate phosphorylation in (C) HCT116 cells treated with 500 μM phosphodiesterase inhibitor IBMX and 10 μM adenylyl cyclase activator forskolin (F), or treated with 10 μM ONC201, in the presence or absence of 1 μM protein kinase A inhibitor peptide 6-22 (PKI); (D) Hep3B cells treated with different doses of ONC201 for 24 hours; (E) HCT116, RKO, and ONC201-resistant RKO cells (RKOr1) treated with 10 μM (HCT116) or 5 μM (RKOp and RKOr1) ONC201 for indicated times.

Article Snippet: Five different lentiviral plasmid shRNA constructs for DRD2 (Origene) were amplified, and DNA was isolated using plasmid mini kit (Qiagen) according to manufacturer's instructions.

Techniques: Functional Assay, Expressing, Western Blot, Phospho-proteomics

Journal: Neoplasia (New York, N.Y.)

Article Title: Role of Dopamine Receptors in the Anticancer Activity of ONC201.

doi: 10.1016/j.neo.2017.10.002

Figure Lengend Snippet: Figure 3. Transient knockdown of DRD2 in colorectal cancer cells can activate the ISR and reduce cell number. (A) Viable cell count, (B) Western blot analyses of PARP cleavage and CHOP expression, (C) Western blot analyses of ATF4 and CHOP protein expression in cells after DRD2 siRNA-mediated knockdown for 24 hours and subsequent treatment with 10 μM ONC201 for indicated times. *P b .05 versus vehicle-treated scramble siRNA- transfected; P b .05 versus vehicle-treated DRD2 siRNA-transfected. (D) qRT-PCR analyses of DRD1 and DRD2 mRNA expression were performed to verify knockdown and monitor for potential compensatory overexpression of DRD1 receptor. Data are means ± SE from three biological replicates. P b .05 versus viability of control shRNA cells similarly treated. (E) Western blot analyses for PKA substrate phosphorylation in stably transfected control and DRD2 shRNA cells treated with 10 μM ONC201 for 24 hours. (F) Cell proliferation rate assessment of control and DRD2 shRNA-transfected cells was performed by enumerating cell number after indicated times of cell culture. Data are means ± SE from three biological replicates. P b .05 versus viability of control shRNA cells similarly treated.

Article Snippet: Five different lentiviral plasmid shRNA constructs for DRD2 (Origene) were amplified, and DNA was isolated using plasmid mini kit (Qiagen) according to manufacturer's instructions.

Techniques: Knockdown, Cell Counting, Western Blot, Expressing, Transfection, Quantitative RT-PCR, Over Expression, Control, shRNA, Phospho-proteomics, Stable Transfection, Cell Culture

Journal: Neoplasia (New York, N.Y.)

Article Title: Role of Dopamine Receptors in the Anticancer Activity of ONC201.

doi: 10.1016/j.neo.2017.10.002

Figure Lengend Snippet: Figure 4. The anticancer effects of ONC201 are distinct from other selective DRD2 antagonists. (A) CTG assay images, (B) apoptosis measured by sub-G1 analyses, and Western blot analyses of (C) PARP cleavage and (D) ATF4 protein expression in different cancer cell types treated with different selective DRD2 antagonists (O: 10 μM ONC201 for HCT116, SKNSH, and U251, and 5 μM ONC201 for RKO; L: 25 μM L741626; R: 10 μM). (E) Nontumorigenic CCD-CON and IMR90 cells and HCT116 and SKNSH cancer cells treated with indicated doses of L-741,626 for 72 hours. *P b .05 versus vehicle treated; P b .05 versus L-741,626 treated.

Article Snippet: Five different lentiviral plasmid shRNA constructs for DRD2 (Origene) were amplified, and DNA was isolated using plasmid mini kit (Qiagen) according to manufacturer's instructions.

Techniques: CTG Assay, Western Blot, Expressing

Journal: bioRxiv

Article Title: Modulation of huntingtin S421 phosphorylation in a Huntington’s disease mouse model and its detection in nonhuman primate cerebrospinal fluid

doi: 10.1101/2025.07.18.665500

Figure Lengend Snippet: A. Specificity assessment of Ab2174 for pS421 HTT by WB and the SMC assay in purified, recombinant FLQ23 HTT proteins with/without S421 A or D mutations . i) WB probed with Ab2174 polyclonal antibody or MAB2166 antibody reveals pS421-HTT and total HTT levels, respectively. ii) SMC analysis of pS421-HTT levels (MW1-Ab2174) or total HTT levels (MW1-2B7) performed on serial dilutions of recombinant proteins in i). Representative experiment of n=3 is shown. B. Specificity assessment of Ab2174 for pS421 HTT by WB and SMC assay in HEK293T cells overexpressing FL HTT with/without S421 A or D mutations. i) WB probed with Ab2174 polyclonal antibody or MAB2166 antibody to demonstrate the levels of pS421-HTT and total HTT, respectively. Anti-GAPDH was used as loading control. ii) Normalized pS421 signal (MW1-Ab2174 SMC assay) on total HTT levels (MW1-2B7 SMC assay) performed on samples in i). Means and standard deviations from n=3 biological replica. C. Pharmacological modulation of pS421-HTT levels in HEK293T cells transfected with N571 Q55 HTT protein and FLAG-tagged dopamine D2 receptor, treated with D2R agonist (Bromocriptine) or antagonists (Haloperidol, Raclopride). i) WB probed with MAB2166 antibody, Ab2174 polyclonal antibody or FLAG antibody demonstrating total HTT levels, pS421-HTT and the D2 receptor, respectively. Anti-GAPDH used as loading control. ii) Densitometric analysis of WB in i) and normalized pS421 signal (MW1-Ab2174 SMC assay) on total HTT levels (MW1-2B7 SMC assay) performed on the same samples. Means and standard deviations from n=3 biological replica. Statistical analysis by one-way analysis of variance (*P<0.05; **P < 0.01; ***P < 0.005).

Article Snippet: The pCMV6 expression plasmids containing the cDNA coding for C-term Myc-FLAG-tagged dopamine D2 receptor (D2R; catalog #RC213065) and Protein kinase A catalytic subunit (PKA C α ) (PRKACA; catalog #RC210332) were obtained from ORIGENE (Rockville, Maryland, United States).

Techniques: Purification, Recombinant, Control, Transfection

Journal: Frontiers in Cellular Neuroscience

Article Title: Inhibitory Effects of Trifluoperazine on Peripheral Proinflammatory Cytokine Expression and Hypothalamic Microglia Activation in Obese Mice Induced by Chronic Feeding With High-Fat-Diet

doi: 10.3389/fncel.2021.752771

Figure Lengend Snippet: List of reagents (antibodies and assay kits) in the study.

Article Snippet: Polyclonal rabbit anti-D2R , Biorbyt, Cat# orb10515, RRID:AB_10747533.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Adenylyl cyclases combinatorially integrate opposing dopamine receptor signals

doi: 10.64898/2026.07.10.737756

Figure Lengend Snippet: (A, B). Single-cell dose-response curves for (A) Gαs-coupled DRD1 and (B) Gαi-coupled DRD2 in CHO-K1::cAMPinG1 cells transiently expressing HALO-tagged receptors. DRD2 was stimulated in the presence of 10 µM forskolin to elevate the baseline cAMP level. Cells are binned and colored by their measured surface receptor abundance (arbitrary flow-cytometry units; see color bars). Solid lines represent joint fits of the operational model (R 2 = 0.98 for DRD1; R 2 = 0.97 for DRD2) with binding affinity (Kd) locked to established IUPHAR values (fit values and 95% CIs are provided in ). Data represent per-bin medians, with error bars indicating ±1 SEM across independent replicate dates (n = 4 for DRD1, n = 5 for DRD2). The remaining subtypes (DRD3, DRD4, DRD5) are characterized in Fig. S3. (C, D) DRD1 dopamine dose-response under siRNA knockdown of Gαs at (C) 1 nM and (D) 33 nM siGNAS (ΔGs), overlaid on the wild-type Gαs response, binned by surface DRD1 abundance (colors as in A). Solid lines are the shared-parameter operational-model fit in which only the available transducer pool [Gs] varies per condition ; Gαs depletion right-shifts the curves along the dose axis without lowering the maximal response. (E). Schematic of the modified operational model. Ligand binding ([L]) yields a fractional receptor occupancy (ρ). This fraction is multiplied by the measured surface receptor abundance ([R]) to yield a composite signaling drive (D = [R] · [G] · ρ). The drive is processed through a saturable Hill-type transducer function (S) to compute the final integrated cAMP output (y). Full mathematical derivation is provided in the Supplementary Information. (F). Master-curve collapse across receptor abundances. Plotting the normalized transducer output (S) against the composite signaling drive (D) collapses the variable dose-response families onto a single, invariant master curve for each receptor subtype. This confirms that cellular cAMP responses are governed by the combined drive quantity rather than receptor density or ligand concentration independently.

Article Snippet: For experiments involving Gαi-coupled receptors (DRD2, DRD3, DRD4), adenylyl cyclase was activated with 10 μM forskolin (MedChemExpress, HY-15371) 5 minutes post-dopamine treatment.

Techniques: Single Cell, Expressing, Flow Cytometry, Binding Assay, Knockdown, Modification, Ligand Binding Assay, Concentration Assay

Journal: bioRxiv

Article Title: Adenylyl cyclases combinatorially integrate opposing dopamine receptor signals

doi: 10.64898/2026.07.10.737756

Figure Lengend Snippet: (A–C) Single-cell dose-response curves for (A) Gαo-coupled DRD3, (B) Gαi-coupled DRD4, and (C) Gαs-coupled DRD5 in CHO-K1::cAMPinG1 cells transiently expressing HALO-tagged receptors. The Gαi/o-coupled subtypes (DRD3, DRD4) were stimulated in the presence of 10 µM forskolin to elevate the baseline cAMP level; the Gαs-coupled DRD5 was measured from the unstimulated baseline. Cells are binned and colored by their measured surface receptor abundance (arbitrary flow-cytometry units; see color bar). Solid lines represent fits of the operational model with binding affinity (Kd) locked to established IUPHAR values (fit values and 95% CIs are provided in ). Data represent per-bin medians, with error bars indicating ±1 SEM across independent replicate dates (n = 2 for DRD3, n = 3 for DRD4, n = 4 for DRD5). (A) DRD3 produces no dopamine-evoked cAMP response, because its primary transducer Gαo is not endogenously expressed in CHO-K1 ; it therefore serves as a silent control. (B) DRD4 shows shallow, expression-dependent suppression of the forskolin-elevated baseline (R 2 = 0.91), consistent with DRD4’s canonical preference for Gαo over Gαi. (C) DRD5 shows expression-dependent cAMP stimulation (R 2 = 0.97). These subtypes are reported for completeness of the receptor panel and are not used in any quantitative downstream analysis; the identifiability of their fits is examined in .

Article Snippet: For experiments involving Gαi-coupled receptors (DRD2, DRD3, DRD4), adenylyl cyclase was activated with 10 μM forskolin (MedChemExpress, HY-15371) 5 minutes post-dopamine treatment.

Techniques: Single Cell, Expressing, Flow Cytometry, Binding Assay, Control

Journal: bioRxiv

Article Title: Adenylyl cyclases combinatorially integrate opposing dopamine receptor signals

doi: 10.64898/2026.07.10.737756

Figure Lengend Snippet: (A) Per-receptor scatter of dopamine binding measurements before and after the stringent curation that produced . Each colored point (orange = Gαs-coupled, blue = Gαi-coupled) is one of the 24 measurements retained in the main figure; each light-grey point is a measurement excluded by one or more curation criteria (see Methods and panel B of this figure). All rows that map to one of the five dopamine receptors are included, with descriptions like “D2L”, “D2A”, and “D2long” mapped to DRD2 (long isoform) by an extended description-matching rule applied in this SI figure but not in the main-text curation, in order to expose the full per-receptor scatter that the database contains. n (kept/total): DRD1 = 6/39, DRD5 = 1/2, DRD2 = 5/116, DRD3 = 10/50, DRD4 = 2/21. Solid colored bars, stringent median (matches ); dashed black ticks, unfiltered median across all rows shown. The two medians lie within 0.1 log unit (≈25%) of each other for every receptor that has more than one entry, indicating that the stringent filter narrows the database but does not systematically bias the central tendency. (B) Counts of GtoPdb rows excluded by each curation criterion. Each row is assigned to the first applicable category in the order: functional assay (Assay Type = “F” in GtoPdb; signaling readouts such as cAMP, β-arrestin recruitment, or [ S]GTPγS binding rather than direct ligand binding), species not specified (assay description does not explicitly state the species), non-human species (description names rat, mouse, bovine, dog, or porcine receptor), other / multiple (combinations of the above or unparseable values, value reported as a range, or “unknown origin” phrase in description), unrecognized assay type (Assay Type column does not contain the standard B/F values).

Article Snippet: For experiments involving Gαi-coupled receptors (DRD2, DRD3, DRD4), adenylyl cyclase was activated with 10 μM forskolin (MedChemExpress, HY-15371) 5 minutes post-dopamine treatment.

Techniques: Binding Assay, Produced, Functional Assay, Ligand Binding Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: Nucleus accumbens shell modulates seizure propagation in a mouse temporal lobe epilepsy model

doi: 10.3389/fcell.2022.1031872

Figure Lengend Snippet: Increased excitability of D1R- and D2R-MSNs in the NAc shell after TLE seizures. (A) Schematic diagram of virus microinjection to enable expression of eGFP reporter on D1R-MSNs or D2R-MSNs in the NAc shell and KA microinjection into BLA to induce TLE seizures of C57BL/6 mouse. (B) Schematic diagram of TLE seizure induction followed by in intro electrophysiological recording in the NAc shell. (C) Representative current-clamp recordings. (D) Summary input–output curves of D1R-MSN of the mice treated KA ( n = 15 cells) show increased AP generations in response to current injections, comparing with the control mice treated with ACSF ( n = 18 cells). *** p < 0.001, F = 70.049, df = 1, two-way ANOVA with Tukey post-hoc test. (E–I) Summary data of resting membrane potential (RMP), AP threshold, rheobase current, and membrane input resistance (R in ) of D1R-MSN of the mice treated with ACSF ( n = 18 cells) or KA ( n = 15 cells). ** p < 0.01, *** p < 0.001, Student’s unpaired t-test. (J) Representative current-clamp recordings and (K) summary input–output curves of D2R-MSN of the mice treated KA ( n = 13 cells) show increased AP generations in response to current injections, compared with the control mice treated with ACSF ( n = 15 cells). *** p < 0.001, F = 20.136, df = 1, two-way ANOVA with Tukey post-hoc test. (L–O) Summary data of resting membrane potential (RMP), AP threshold, rheobase current, and membrane input resistance (R in ) of D2R-MSN of the mice treated with ACSF ( n = 15 cells) or KA ( n = 13 cells). ** p < 0.01, *** p < 0.001, student’s unpaired t-test.

Article Snippet: In the experiments investigating functional interaction between D1R-MSN and D2R-MSN, 15 μM SKF 38393 hydrochloride (MedChemExpress #HY-12520 A) was used to activate D1R, 15 μM quinpirole hydrochloride (sigma # Q102) was used to activate D2R, and 100 μM CGP35348 (MedChemExpress #HY-103530) was used to block GABA B receptors.

Techniques: Virus, Microinjection, Expressing, Control, Membrane

Journal: Frontiers in Cell and Developmental Biology

Article Title: Nucleus accumbens shell modulates seizure propagation in a mouse temporal lobe epilepsy model

doi: 10.3389/fcell.2022.1031872

Figure Lengend Snippet: Increased synaptic transmission of D1R-MSN, but not D2R-MSN, in the NAc shell after TLE seizures. (A) Representative sEPSC of D1R-MSN of the mice treated with ACSF or KA. (B) Cumulative probability of the amplitudes and the averaged amplitude of sEPSCs of D1R-MSN of the mice treated with ACSF ( n = 9 cells) or KA ( n = 14 cells). * p < 0.05, Student’s unpaired t-test (the same test was used below in this figure). (C) Cumulative probability of the inter-event intervals of sEPSCs and the averaged frequency of sEPSCs of D1R-MSN of the mice treated with ACSF ( n = 14 cells) or KA ( n = 9 cells). *** p < 0.001. (D) Representative recordings of sIPSC of D1R-MSN of the mice treated with ACSF or KA. (E) Cumulative probability of the amplitudes and the averaged amplitude of sIPSCs of D1R-MSN of the mice treated with ACSF ( n = 9 cells) or KA ( n = 8 cells). (F) Cumulative probability of the inter-event intervals of sIPSCs and the averaged frequency of sIPSCs of D1R-MSN of the mice treated with ACSF ( n = 9 cells) or KA ( n = 8 cells). (G) Representative recordings of sEPSC of D2R-MSN of the mice treated with ACSF or KA. (H) Cumulative probability of the amplitudes and the averaged amplitude of sEPSCs of D2R-MSN of the mice treated with ACSF ( n = 10 cells) or KA ( n = 8 cells). (I) Cumulative probability of the inter-event intervals of sEPSCs and the averaged frequency of sEPSCs of D2R-MSN of the mice treated with ACSF ( n = 10 cells) or KA ( n = 8 cells). (J) Representative recordings of sIPSC of D2R-MSN of the mice treated with ACSF or KA. (K) Cumulative probability of the amplitudes and the averaged amplitude of sIPSCs of D2R-MSN of the mice treated with ACSF ( n = 10 cells) or KA ( n = 10 cells). (L) Cumulative probability of the inter-event intervals of sIPSCs and the averaged frequency of sIPSCs of D2R-MSN of the mice treated with ACSF ( n = 10 cells) or KA ( n = 10 cells).

Article Snippet: In the experiments investigating functional interaction between D1R-MSN and D2R-MSN, 15 μM SKF 38393 hydrochloride (MedChemExpress #HY-12520 A) was used to activate D1R, 15 μM quinpirole hydrochloride (sigma # Q102) was used to activate D2R, and 100 μM CGP35348 (MedChemExpress #HY-103530) was used to block GABA B receptors.

Techniques: Transmission Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: Nucleus accumbens shell modulates seizure propagation in a mouse temporal lobe epilepsy model

doi: 10.3389/fcell.2022.1031872

Figure Lengend Snippet: Chemogenetic inhibition of D1R-MSN or D2R-MSN in the NAc shell alleviates TLE seizures. (A) Representative recording shows spontaneous AP firings of D1R-MSN and (B) D2R-MSN transfected with DREADD receptor hM4D (Gi) are suppressed by CNO (5 μM) perfusion. Membrane potential was held under current-clamp at around −50 mV. (C) Representative confocal images show the expression of mCherry and eGFP on D1R-MSN transfected with AAV2/9-DIO-hM4D(Gi)-mCherry and AAV2/9-D1-Cre-eGFP viruses. (D) Representative confocal images show the expression of mCherry and eGFP on D2R-MSN transfected with AAV2/9-DIO-hM4D(Gi)-mCherry and AAV2/9-D2-Cre-eGFP viruses. (E) Schematic diagram of experimental design to examine the effects of chemogenetic inhibition of D1R-MSN or D2R-MSN in the NAc shell on TLE seizures induced by intra-basolateral amygdala (intra-amygdala) microinjection of KA. The mice are allocated into four groups: KA group n = 13 mice; CNO + KA group n = 14 mice; CNO + D1-Gi + KA group n = 10 mice; CNO + D2-Gi + KA group n = 15 mice. (F) Racine’s scaling for maximal seizure stage every 10 min during the whole course of 120 min after KA microinjection. * p < 0.05, *** p < 0.001, F = 18.46, df = 3, two-way ANOVA with repeated-measures. (G) Onset time of the first seizure after KA microinjection. One-way ANOVA. (H) Latency to secondary GSs after KA microinjection. ** p < 0.01, one-way ANOVA. (I) Total number of secondary GSs after KA microinjection. *** p < 0.001, one-way ANOVA. (J) Maximal seizure stage in the whole course of 120 min after KA microinjection. ** p < 0.01, Kruskal–Wallis H test with Bonferroni correction (K) Mortality rate after KA microinjection. p > 0.05, Kruskal–Wallis H test. (L) Representative recording and statistical data (M) show AP firing in response to a step current before or after perfusion of 15 μM D1R agonist SKF 38393. n = 21 cells, * p < 0.05, Student’s unpaired t-test. (N) Representative recording and statistical data (O) show AP firing in response to a step current before or after perfusion of 15 μM D2R agonist quinpirole. n = 18 cells, * p < 0.05, Student’s unpaired t-test. (P) Representative recording and statistical data (Q) show AP firing in response to a step current before or after perfusion of 15 μM D1R agonist SKF 38393 in the presence of GABA A/B blockers (10 μM SR95531/100 μM CGP35348). n = 15 cells, p > 0.05, Student’s unpaired t-test. (R) Representative recording and statistical data (S) show AP firing in response to a step current before or after perfusion of 15 μM D2R agonist quinpirole in the presence of GABA A/B blockers. n = 20 cells, p > 0.05, Student’s unpaired t-test. In the figures G–K, the statistical differences are compared between the KA group and the CNO + KA group, and between the CNO + KA group and the CNO + D1R-Gi + KA group or the CNO + D2R-Gi + KA group. n.s means not statistically significant.

Article Snippet: In the experiments investigating functional interaction between D1R-MSN and D2R-MSN, 15 μM SKF 38393 hydrochloride (MedChemExpress #HY-12520 A) was used to activate D1R, 15 μM quinpirole hydrochloride (sigma # Q102) was used to activate D2R, and 100 μM CGP35348 (MedChemExpress #HY-103530) was used to block GABA B receptors.

Techniques: Inhibition, Transfection, Membrane, Expressing, Microinjection