성상교세포

Nature 616권, 764~773페이지(2023)이 기사 인용

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성상세포와 뉴런은 뇌에서 광범위하게 상호작용합니다. 성상교세포와 뉴런 프로테옴을 식별하는 것은 생리학과 질병에 대한 각각의 기여를 결정하는 단백질 네트워크를 밝히는 데 필수적입니다. 여기에서 우리는 생체 내에서 선조체 성상교세포와 뉴런의 프로테옴을 연구하기 위해 세포 특이적 및 하위 구획 특이적 근접 의존성 biotinylation1을 사용했습니다. 우리는 성상교세포와 뉴런의 세포질과 원형질막 구획을 평가하여 이들 세포가 신호 전달 기계의 단백질 수준에서 어떻게 다른지 발견했습니다. 우리는 또한 성상교세포의 하위 프로테옴과 필수 성상교세포 신호 전달 및 항상성 기능의 분자적 기초를 밝히기 위해 말단 발과 미세 돌기를 포함한 성상교세포의 세포내 구획을 평가했습니다. 특히, 강박 장애(OCD) 및 반복적 행동2,3,4,5,6,7,8과 관련된 SAPAP3(Dlgap3에 의해 인코딩됨)은 선조체 성상교세포에서 높은 수준으로 검출되었으며 특정 성상교세포 내에 풍부했습니다. 액틴 세포골격 조직을 규제하는 하위 구획. 또한, SAPAP3가 결여된 OCD4의 마우스 모델에서 행동 분석 및 분자 평가와 결합된 유전자 구조 실험은 반복적이고 불안과 관련된 OCD 유사 표현형에 대한 성상 세포 및 신경 세포 SAPAP3의 뚜렷한 기여를 보여주었습니다. 우리의 데이터는 성상교세포와 뉴런이 단백질 수준과 주요 신호 전달 경로에서 어떻게 다른지 정의합니다. 더욱이, 그들은 성상교세포 하위단백질체가 어떻게 생리학적 하위구획 사이에서 달라지는지, 그리고 성상교세포와 신경 SAPAP3 메커니즘이 생쥐의 OCD 표현형에 어떻게 기여하는지를 밝혔습니다. 우리의 데이터는 성상교세포와 뉴런 모두를 표적으로 하는 치료 전략이 OCD 및 잠재적으로 다른 뇌 장애를 탐색하는 데 유용할 수 있음을 나타냅니다.

성상교세포는 중추신경계에서 지배적인 유형의 신경교이며 뉴런과 함께 진화해 왔습니다. 성상교세포는 뇌10의 중요한 구성 요소이며 뉴런과 마찬가지로 뇌 영역마다 다른 형태와 특성을 나타냅니다. 성상교세포와 뉴런 모두 정신 장애를 포함한 뇌 질환15에 광범위하게 연루되어 있습니다. 그러나 공유되거나 분리된 성상세포 및 신경 분자 메커니즘과 생쥐의 정의된 정신 질환 또는 표현형과 관련된 뇌 영역 내에서의 각각의 기여에 대해서는 알려진 바가 거의 없습니다.

뉴런과 성상세포는 선조체 내를 포함하여 해부학적, 생리학적으로 상호 작용합니다. 생리학 및 질병 설정에서 대부분의 연구는 신경병리학적 방법, 생리학, 세포 마커 또는 RNA 발현 분석을 사용하여 성상교세포와 뉴런을 비교했습니다. RNA와 관련하여, RNA 발현 수준과 단백질 수준18 사이의 관계는 매우 중요하지만 매우 복잡합니다. 따라서 뉴런과 성상교세포에 대한 특정 단백질 기반 메커니즘을 식별하는 것이 중요합니다. 또한 성상교세포와 뉴런의 기본 생물학을 이해하려면 형태학적으로 손상되지 않은 세포 내에서 단백질의 정체성과 차이점을 포착하는 것이 필요합니다. 세포 해리 및 형광 활성화 세포 분류(FACS) 절차는 대부분의 성상교세포 및 신경 과정을 전단하고 특히 조직 스트레스20,21,22에 반응하는 성상교세포에 손상을 주어 프로테오믹스에 대한 이러한 방법의 사용을 방해합니다. 결과적으로 성상교세포와 뉴런의 프로테옴은 모든 종의 생리학 또는 정신 질환의 관련 표현형에 대한 기여도를 이해하기 위해 직접 측정, 비교 또는 활용되지 않았습니다.

선조체는 운동, 행동 및 다양한 신경정신과적 상태에 관여하는 피질하 핵 그룹인 기저핵의 가장 큰 핵입니다. 선조체에는 성상교세포와 뉴런 사이의 광범위한 접촉이 포함되어 있으며, 그 중 95%는 DARPP32 양성 중간 가시 뉴런(MSN)입니다. 성상교세포는 해리 후 복잡한 형태를 잃기 때문에(확장 데이터 그림 1a-d), 유전적으로 표적화된 비오틴 리가제를 사용하여 세포 유형별 프로테옴(성상교세포 및 뉴런)과 구획(세포질 및 원형질막(PM))의 구성을 특성화했습니다. (BioID2; 확장 데이터 그림 2a, b)는 아데노 관련 바이러스 (AAV, 확장 데이터 그림 2b, c)를 사용하여 선조체 내에서 생체 내 전달되었습니다. 이 방법은 세포 해리나 FACS를 사용하지 않습니다. BioID2는 비오틴이 있는 경우 유리 라이신 잔기의 단백질을 비오티닐화합니다1,26. HEK-293 세포에서 세포질 BioID2 및 PM 표적화 LCK-BioID2 구성체를 특성화한 후(확장 데이터 그림 3), 절단된 GFAP 프로모터(GfaABC1D) 또는 인간 SYN1 프로모터를 사용하여 BioID2 또는 LCK-BioID2를 성상교세포 또는 뉴런에 선택적으로 전달했습니다. 각각 선호하는 성상교세포(Astro) 또는 뉴런(Neuro) 향성을 갖는 AAV(그림 1a 및 확장 데이터 그림 2c-h). Astro BioID2, Astro LCK-BioID2 및 이들이 비오티닐화한 단백질은 끝 발 내부를 포함하여 S100β 양성 부시 성상교세포에서만 검출되었습니다(확장 데이터 그림 4a-d). 반대로, Neuro BioID2, Neuro LCK-BioID2 및 이들의 비오티닐화 단백질은 DARPP32 양성 신경 세포체 및 신경필 내에서 검출되었으며, 이는 각각 축삭 및 수지상 발현을 반영합니다(확장 데이터 그림 4e-h). Western blot 분석을 통해 비오틴화(확장 데이터 그림 2e-h 및 4, 각 경우 P < 0.01)가 확인되었으며, 이는 액체 크로마토그래피-탠덤 질량 분석법(LC-MS/MS)을 통해 단백질 식별이 가능해졌습니다.

1 and FDR < 0.05 versus GFP controls). c, UpSet plot of BioID2-identified proteins. d, LFQ comparison of proteins detected by cytosolic Astro BioID2 and Neuro BioID2. Top, proteins specific to Neuro BioID2 or Astro BioID2 when compared with each other. The four most abundant proteins are named. Bottom, comparison of proteins that were shared in both cytosolic Astro BioID2 and Neuro BioID2. The five highest enriched proteins (log2(FC) > 2) are indicated. The top three proteins that showed no enrichment in either cell are depicted in red. e, As in d but for PM Astro BioID2 and Neuro BioID2. f, Left, STRING analysis map of the top 100 proteins identified with Astro BioID2 and Astro LCK–BioID2. Node size represents the enrichment of each protein versus the GFP control. Edges represent putative interactions from STRING. Right, small clustergrams show categories for biological process. PPI, protein–protein interaction. g. As in f, but for Neuro BioID2 and Neuro LCK–BioID2. h, Expression levels (LFQ intensity) of Ca2+-dependent vesicle release proteins identified by each BioID2 construct. BPs, binding proteins. i, Expression levels of proteins related to lipid metabolism identified in each BioID2 construct./p> 1 and FDR < 0.05 versus GFP controls). The top two most abundant proteins for each subcompartment are named. d, Label-free based quantification comparison of proteins detected in the cytosolic Astro BioID2 and PM Astro LCK–BioID2. Top, specific LCK–BioID2 proteins compared to cytosol. The top four most abundant proteins for LCK–BioID2 are indicated. Bottom, volcano plot comparing proteins that were shared in both cytosolic BioID2 and LCK–BioID2. The five highest enriched proteins for LCK–BioID2 (log2(FC) > 2) are indicated. Magenta label shows protein that was validated by IHC in Extended Data Fig. 11. Red label shows that SAPAP3 is enriched in the astrocyte PM. e,f, As in d but for cytosolic Astro BioID2 and Astro EZR–BioID2 (e) and cytosolic Astro BioID2 and Astro AQP4–BioID2 (f). g, STRING analysis map of the top 50 (by LFQ abundance) biotinylated proteins identified in astrocyte fine processes with Astro EZR–BioID2. Node size represents the enrichment of each protein versus the GFP control. Edges represent putative interactions from the STRING database. h, As in g but for proteins identified in the astrocyte end foot with Astro AQP4–BioID2. i, Bars show the most significant Enrichr gene ontology (GO) term for the unique and enriched proteins found in each astrocyte subcompartment. Top, the GO term for biological process. Bottom, the GO term for molecular function./p>

1 and FDR < 0.05 versus GFP controls were considered putative hits and used for subsequent comparison between subcompartments and cell types. A comparison between subcompartments and cell types was also performed with limma utilizing the same thresholds (log2(FC) > 1 and FDR < 0.05). To account for variations in pull-down efficiency, all proteins and their LFQ values were normalized to pyruvate carboxylase (UniProt identifier Q05920). Downstream analysis was conducted only on proteins with non-zero LFQ values in three or more experimental replicates. Data analysis for whole bulk tissue analyses was carried out in an identical manner, except samples were normalized by median intensity./p> 5 in at least 4 samples per condition and log2(FC) > 1 or < −1 using the Bioconductor package limmaVoom (v.3.36) with the FDR threshold set at <0.05. Differentially expressed genes that were more than twofold higher in the immunoprecipitated samples than the input samples were designated as astrocyte-enriched or neuron-enriched differentially expressed genes. RNA-seq data have been deposited within the Gene Expression Omnibus repository (https://www.ncbi.nlm.nih.gov/geo) with the accession identifier GSE184773./p> 0) in our mouse RNA-seq studies./p>

1 versus GFP controls). The mean LFQ value and SEM are shown. b. Pie chart of PANTHER pathway analysis terms for "biological processes". Pie chart shows the number of proteins found for each term from the 332 Astro BioID2 proteins. c. As in b, but for the 434 Neuro BioID2 proteins. d. As in b, but for 310 Astro Lck-BioID2 proteins. e. As in b, but for the 1672 Neuro Lck-BioID2 proteins. f. Bar graph denotes the number of calcium dependent vesicle release protein isoforms detected in each BioID2 construct experiment. g. Bar graph denotes the number of lipid metabolism related proteins that were detected in each BioID2 construct experiment./p> 1 versus GFP controls). The mean LFQ value and SEM are shown. For blot source data, see Supplementary Fig. 1./p> 1 and FDR < 0.05 versus GFP controls) detected in the cytosolic Astro BioID2 and plasma membrane Astro Lck-BioID2 reveal plasma membrane enriched proteins. Top half of the volcano plot shows 238 unique Lck-BioID2 proteins when compared to cytosol. The top four most abundant proteins for Lck-BioID2 are shown. Lower half of volcano plot shows comparison of 144 proteins that were common in both cytosolic BioID2 and Lck-BioID2. The five highest enriched proteins for Lck-BioID2 (Log2FC > 2) are shown. Magenta label shows protein that was validated with IHC in panel d. Red label shows Dlgap3/SAPAP3 is enriched in the astrocyte plasma membrane. c. Heat map shows the rank-rank hypergeometric overlap (RRHO) of the RNA and protein rank for the 270 plasma membrane proteins. Each pixel represents the significance of overlap between the two datasets in –log10(P-value). Red pixels represent highly significant overlap. Color scale denotes the range of P-values at the negative log10 scale (bin size = 100). d. IHC analysis of Slc4a4 (Nbc1) protein in tdTomato and Lck-GFP labeled astrocytes shows co-localization within the astrocyte territory. Scale bar represents 20 μm. e. Co-localization analysis using Pearson's r co-efficient shows high co-localization between Lck-GFP and Slc4a4 (Nbc1). The mean and SEM are shown (n = 8 tdTomato+ cells from 4 mice; Two-tailed paired t-test). f. Scale-free STRING analysis protein-protein association map of the 270 unique and enriched biotinylated proteins identified in astrocyte plasma membrane with Astro Lck-BioID2 . Node size represents the enrichment of each protein vs the GFP control (log2(BioID2/GFP)). Edges represent putative interactions from the STRING database. Bar graphs show the functional enrichment analysis of the 270 proteins using "Biological process", "Cellular component", and "Molecular function" terms from Enrichr./p> 1 and FDR < 0.05 versus GFP controls) detected in the cytosolic Astro BioID2 and end foot Astro Aqp4-BioID2 reveal end foot enriched proteins. Top half of the volcano plot shows 577 unique Aqp4-BioID2 proteins when compared to cytosol. The top four most abundant proteins for Aqp4-BioID2 are shown. Lower half of volcano plot shows comparison of 228 proteins that were common in both cytosolic BioID2 and Aqp4-BioID2. The five highest enriched proteins for Aqp4-BioID2 (Log2FC > 2) are shown. Magenta label shows protein that was validated with immunohistochemistry in panel d. c. Heat map shows the rank-rank hypergeometric overlap (RRHO) of the RNA and protein rank for the 635 endfoot proteins. Each pixel represents the significance of overlap between the two datasets in –log10(P-value). Red pixels represent highly significant overlap. Color scale denotes the range of P-values at the negative log10 scale (bin size = 100). d. IHC analysis of PAICS protein in tdTomato and Lck-GFP labeled astrocytes shows co-localization within the astrocyte territory. Scale bar represents 20 μm. e. Co-localization analysis using Pearson's r co-efficient shows high co-localization between Aqp4-GFP and PAICS. The mean and SEM are shown (n = 8 tdTomato+ cells from 4 mice; Two-tailed paired t-test). f. Scale-free STRING analysis protein-protein association map of the top 100 unique and enriched biotinylated proteins identified in the astrocyte endfoot with Astro Aqp4-BioID2 . Node size represents the enrichment of each protein vs the GFP control (log2(BioID2/GFP)). Edges represent putative interactions from the STRING database. Bar graphs show the functional enrichment analysis of the 635 proteins using "Biological process", "Cellular component", and "Molecular function" terms from Enrichr. The image of the astrocyte subcompartments in panel a was created using BioRender (https://www.biorender.com/)./p> 1 and FDR < 0.05 versus GFP controls) detected in the cytosolic Astro BioID2 and fine process Astro Ezrin-BioID2 reveal fine process enriched proteins. Top half of the volcano plot shows 216 unique Ezrin-BioID2 proteins when compared to cytosol. The top four most abundant proteins for Ezrin-BioID2 are shown. Lower half of volcano plot shows comparison of 186 proteins that were common in both cytosolic BioID2 and Ezrin-BioID2. The five highest enriched proteins for Ezrin-BioID2 (Log2FC > 2) are shown. Magenta label shows protein that was validated with immunohistochemistry in panel d. Red label shows Dlgap3/SAPAP3 is enriched in the astrocyte fine processes. c. Heat map shows the rank-rank hypergeometric overlap (RRHO) of the RNA and protein rank for the 234 Ezr-BioID2 proteins. Each pixel represents the significance of overlap between the two datasets in –log10(P-value). Red pixels represent highly significant overlap. Color scale denotes the range of P-values at the negative log10 scale (bin size = 100). d. IHC analysis of Nebl protein in tdTomato and Ezr-GFP labeled astrocytes shows co-localization within the astrocyte territory. Scale bar represents 20 μm. e. Co-localization analysis using Pearson's r co-efficient shows high co-localization between Ezr-GFP and Nebl. The mean and SEM are shown (n = 8 tdTomato+ cells from 4 mice; Two-tailed paired t-test). f. Scale-free STRING analysis protein-protein association map of the 234 unique and enriched biotinylated proteins identified in astrocyte processes with Astro Ezr-BioID2 . Node size represents the enrichment of each protein vs the GFP control (log2(BioID2/GFP)). Edges represent putative interactions from the STRING database. Bar graphs show the functional enrichment analysis of the 234 proteins using "Biological process", "Cellular component", and "Molecular function" terms from Enrichr. The image of the astrocyte subcompartments in panel a was created using BioRender (https://www.biorender.com/)./p> 1 and FDR < 0.05 versus GFP controls) detected in the cytosolic Astro BioID2 and Astro Glt1-BioID2 reveal Glt1 enriched proteins. Top half of the volcano plot shows 527 unique Glt1-BioID2 proteins when compared to cytosol. The top four most abundant proteins for Glt1-BioID2 are shown. Lower half of volcano plot shows comparison of 230 proteins that were common in both cytosolic BioID2 and Glt1-BioID2. The five highest enriched proteins for Glt1-BioID2 are shown. Magenta label shows protein that was validated with immunohistochemistry. c. Heat map shows the rank-rank hypergeometric overlap (RRHO) of the RNA and protein rank for the 532 Glt1-BioID2 proteins. Each pixel represents the significance of overlap between the two datasets in –log10(P-value). Red pixels represent highly significant overlap. Color scale denotes the range of P-values at the negative log10 scale (bin size = 100). d. IHC analysis of Faim2 protein in tdTomato and Glt1-GFP labeled astrocytes shows co-localization within the astrocyte territory. Scale bar represents 20 μm. e. Co-localization analysis using Pearson's r co-efficient shows high co-localization between Glt1-GFP and Faim2. The mean and SEM are shown (n = 8 tdTomato+ cells from 4 mice; Two-tailed paired t-test). f. Scale-free STRING analysis protein-protein association map of the top 100 unique and enriched biotinylated proteins identified with Astro Glt1-BioID2 . Node size represents the enrichment of each protein vs the GFP control (log2(BioID2/GFP)). Edges represent putative interactions from the STRING database. Bar graphs show the functional enrichment analysis of all 532 proteins using "Biological process", "Cellular component", and "Molecular function" terms from Enrichr. The image of the astrocyte subcompartments in panel a was created using BioRender (https://www.biorender.com/)./p> 1 and FDR < 0.05 versus GFP controls) detected in the cytosolic Astro BioID2 and Astro Kir4.1-BioID2 reveal Kir4.1 enriched proteins. Top half of the volcano plot shows 390 unique Kir4.1-BioID2 proteins when compared to cytosol. The top four most abundant proteins for Kir4.1-BioID2 are shown. Lower half of volcano plot shows comparison of 275 proteins that were common in both cytosolic BioID2 and Kir4.1-BioID2. The five highest enriched proteins for Kir4.1-BioID2 are shown. Magenta label shows protein that was validated with immunohistochemistry. c. Heat map shows the rank-rank hypergeometric overlap (RRHO) of the RNA and protein rank for the 393 Kir4.1-BioID2 proteins. Each pixel represents the significance of overlap between the two datasets in –log10(P-value). Red pixels represent highly significant overlap. Color scale denotes the range of P-values at the negative log10 scale (Bin size = 100). d. IHC analysis of Hepacam protein in tdTomato and Kir4.1-GFP labeled astrocytes shows co-localization within the astrocyte territory. Scale bar represents 20 μm. e. Co-localization analysis using Pearson's r co-efficient shows high co-localization between Kir4.1-GFP and Hepacam. The mean and SEM are shown (n = 8 tdTomato+ cells from 4 mice; Two-tailed paired t-test). f. Scale-free STRING analysis protein-protein association map of the top 100 unique and enriched biotinylated proteins identified with Astro Kir4.1-BioID2 . Node size represents the enrichment of each protein vs the GFP control (log2(BioID2/GFP)). Edges represent putative interactions from the STRING database. Bar graphs show the functional enrichment analysis of all 393 proteins using "Biological process", "Cellular component", and "Molecular function" terms from Enrichr. The image of the astrocyte subcompartments in panel a was created using BioRender (https://www.biorender.com/)./p> 1 and FDR < 0.05 versus GFP controls) detected in the cytosolic Astro BioID2 and Astro Cx43-BioID2 reveal Cx43 enriched proteins. Top half of the volcano plot shows 179 unique Cx43-BioID2 proteins when compared to cytosol. The top four most abundant proteins for Cx43-BioID2 are shown. Lower half of volcano plot shows comparison of 116 proteins that were common in both cytosolic BioID2 and Cx43-BioID2. The five highest enriched proteins for Cx43-BioID2 are shown. Magenta label shows protein that was validated with immunohistochemistry. c. Heat map shows the rank-rank hypergeometric overlap (RRHO) of the RNA and protein rank for the 196 Cx43-BioID2 proteins. Each pixel represents the significance of overlap between the two datasets in –log10(P-value). Red pixels represent highly significant overlap. Color scale denotes the range of P-values at the negative log10 scale (bin size = 100). d. IHC analysis of Arpc1a protein in tdTomato and Cx43-GFP labeled astrocytes shows co-localization within the astrocyte territory. Scale bar represents 20 μm. e. Co-localization analysis using Pearson's r co-efficient shows high co-localization between Cx43-GFP and Aprc1a. The mean and SEM are shown (n = 8 tdTomato+ cells from 4 mice; Two-tailed Wilcoxon matched-pairs signed rank test). f. Scale-free STRING analysis protein-protein association map of the 196 unique and enriched biotinylated proteins identified with Astro Cx43-BioID2 . Node size represents the enrichment of each protein vs the GFP control (log2(BioID2/GFP)). Edges represent putative interactions from the STRING database. Bar graphs show the functional enrichment analysis of all 196 proteins using "Biological process", "Cellular component", and "Molecular function" terms from Enrichr. The image of the astrocyte subcompartments in panel a was created using BioRender (https://www.biorender.com/)./p> 1 and FDR < 0.05 versus GFP controls) detected in the Astro BioID2-SAPAP3 and Neuro BioID2-SAPAP3 reveal unique astrocyte and neuron SAPAP3 interactors. Top half of the volcano plot shows 306 unique Neuro BioID2-SAPAP3 proteins and 49 unique Astro BioID2-SAPAP3 proteins when compared to each other. The top four most abundant proteins for each cell type are shown. Lower half of volcano plot shows comparison of 228 proteins that were common in both Astro BioID2-SAPAP3 and Neuro BioID2-SAPAP3. The five highest enriched proteins (Log2FC > 2) for neurons are shown. Proteins that did not pass the enrichment threshold for either cell type are represented in the gray box. d. Schematic shows astrocyte specific HA tagged SAPAP3, GFP fused Ezrin, and GFP fused Glt-1 used in AAV constructs to assess interactions via co-immunoprecipitation. 16 week old wild type mice were injected in the striatum with one of the following combinations: HA-SAPAP3 + Ezr-GFP, HA-SAPAP3 + Glt-1-GFP, HA-SAPAP3 only, Ezr-GFP only, or Glt1-GFP only. Western blot shows the immunoprecipitation of either HA or GFP after protein complex isolation. The band 110 kD represents the HA-SAPAP3 band, while the 90 kD bands represent Ezrin-GFP (93 kD) or Glt1-GFP (92 kD). n = 4 mice per combination, 3 technical replicates. e. Representative images of immunostained mouse striatum injected with astrocyte-specific GFP-SAPAP3 (Astro SAPAP3). Left panel shows the immunostaining pattern with S100β as an astrocyte cell marker and right panel shows the immunostaining pattern with DARPP32 as a neuron cell marker. f. Bar graphs depicting the percent of S100β positive or NeuN positive cells with HA expression in a 20x magnification field of view. Teal portion of the bar graphs show the percent co-localization. Bottom descriptive statistics represent percent of HA+ cells that were not S100β positive as the mean (SD) [SEM] (n = 8 fields of view at 20x magnification from 4 mice) c. Representative images of immunostained mouse striatum injected with astrocyte-specific GFP-SAPAP3 (Astro SAPAP3). Left panel shows the immunostaining pattern with S100β as an astrocyte cell marker and right panel shows the immunostaining pattern with DARPP32 as a neuron cell marker. g. Representative images of immunostained mouse striatum injected with neuron-specific GFP-SAPAP3 (Neuro SAPAP3). Left panel shows the immunostaining pattern with DARPP32 as a neuron cell marker and right panel shows the immunostaining pattern with S100β as an astrocyte cell marker. h. Bar graphs depicting the percent of S100β positive or NeuN positive cells with HA expression in a 20x magnification field of view. Purple portion of the bar graphs show the percent co-localization. Bottom descriptive statistics represent percent of HA+ cells that were not DARPP32 positive as the mean (SD) [SEM] (n = 8 fields of view at 20x magnification from 4 mice). The image of the DNA constructs in panels a and d was created using BioRender (https://www.biorender.com/)./p> 1 and FDR < 0.05) while blue circles show downregulated proteins (Log2FC < 1, FDR < 0.05) in the SAPAP3 KO striatum when compared to wild-type. The top 5 most down regulated and up regulated proteins are shown. Green label shows protein that also appeared in the neuron SAPAP3 interactome. b. List of the 66 proteins that were differentially expressed in the striatum of SAPAP3 KO mice when compared to wild type controls. Heat map and color scale (i) shows the Log2fold change of the 66 proteins versus wild type control. Heat map and color scale (ii) shows the mRNA abundance (FPKM) of the 66 proteins in our neuron or astrocyte specific mouse RNA-seq datasets. Heat map and color scale (iii) shows the Log2fold change at the mRNA level of the 66 proteins in human caudate of OCD subjects compared to controls. Arrowheads show genes that had conserved changes in the mouse SAPAP3 KO model and human OCD. Teal asterisks denote whether the protein was found in the astrocyte specific proteomics datasets, while the purple asterisks denote whether the protein was found in the neuron specific datasets. c. List of proteins shows the 30 significantly (FDR < 0.05) changed genes in human OCD caudate versus control. Heat map depicts the genes’ respective mRNA abundances (FPKM) in our neuron or astrocyte specific mouse RNA-seq datasets. Teal asterisks denote whether the protein was also found in the astrocyte specific proteomics datasets while the purple asterisks denote whether the protein was found in the neuron specific datasets. The gene, C15orf39, was not found in our mouse datasets. d. List of 61 genes associated with OCD and Tourette's syndrome. Heat map depicts the genes’ respective mRNA abundances (FPKM) in our neuron or astrocyte specific mouse RNA-seq datasets. Teal asterisks denote whether the protein was also found in the astrocyte specific proteomics datasets while the purple asterisks denote whether the protein was found in the neuron specific datasets. Orange asterisks denote whether the protein was an astrocytic SAPAP3 interactor, while green asterisks denote whether the protein was a neuronal SAPAP3 interactor./p>

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