Journal: Reproductive biomedicine online

Article Title: Social psychogenic stress promotes the development of endometriosis in mouse.

doi: 10.1016/j.rbmo.2016.11.012

Figure Lengend Snippet: Figure 4 – Immmunoreactivity staining of different markers in ectopic lesions in different groups. (A) Representative immunostaining of ADRB2 in endometrium in CONTROL and SHAM mice and in ectopic endometrium in STRESSED and UNSTRESSED mice. ADRB2 and DRD2 immunoreactivity was both seen primarily in glandular epithelial cells and was localized in the cytoplasm. Scale bar = 125 μm. (B) Representative immunostaining of DRD2, VEGF, CD31, CD41, F4/80, PCNA and α-SMA in the ectopic lesions in UNSTRESSED and STRESSED groups. VEGF immunoreactivity was seen primarily in glandular epithelial cells and was localized in the cytoplasm. CD31 immunostainings were seen mostly in vascular endothelial cells. CD41 shows the extent of platelet aggregation and F4/80 immunoreactivity represents the extent of macrophage infiltration. PCNA immunoreactivity was seen both in glandular epithelial cells and stromal cells were localized in the cell nucleus, but the change of immunoreactivity in glandular epithelial cells was more obvious. α-SMA staining were seen mostly in the stromal component of the ectopic lesions. Scale bar = 125 μm. Mice in stressed (STRS) and unstressed (UNSTRS) groups had undergone endometriosis-inducing surgery, while mice the SHAM group underwent non-endometriosis-inducing surgery. Mice in unstressed (UNSTRS), SHAM and control (CTL) groups were not exposed to stress.

Article Snippet: For negative controls, the immunoglobulin G (IgG) from the rabbit serum (Sigma, Darmstadt, Germany) was used instead of primary antibodies against ADRB2, DRD2, VEGF, CD31, CD41, α-SMA and PCNA except for F4/80, for which the IgG was from the rat serum (Boster, Wuhan, China) was used, all used at the same concentration with its corresponding primary antibody.

Techniques: Staining, Immunostaining, Control

Journal: Brain, behavior, and immunity

Article Title: HIV-1 Tat protein enhances sensitization to methamphetamine by affecting dopaminergic function

doi: 10.1016/j.bbi.2017.05.004

Figure Lengend Snippet: Immunohistochemistry on paraffin embedded sections was utilized examine the protein distribution and levels of dopamine receptor D1 (A, B, C, D), dopamine receptor D2 (E, F, G, H), as well as of IBA-1 (I, J, K, L) in SAL TAT− (A, E, I), SAL TAT+ (B, F, J), METH TAT− (C, G, K), and METH TAT+ (D, H, L) mice. Representative positive cells in the 40× magnification images were labeled with a black arrow. (M) Normalized intensity density was calculated in ImageJ. Data are expressed as Mean ± SEM (n=5). * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Sections were blocked with 5g/l Casein (Sigma Aldrich) in PBS, containing 0.5g/l Thimerosal (Sigma Aldrich) and incubated with Iba-1 antibody (Wako Lab Chemicals, Richmond, VA), the anti-mouse DRD1 antibody (NLS43, Novus Biologicals, Littleton, CO), or anti mouse DRD2 (orb154598, Biorbyt, San Francisco, CA), each one diluted in Casein buffer.

Techniques: Immunohistochemistry, Labeling

Journal: Current biology : CB

Article Title: Transcriptional and anatomical diversity of medium spiny neurons in the primate striatum.

doi: 10.1016/j.cub.2021.10.015

Figure Lengend Snippet: Figure 2. Medium spiny neuron (MSN) subtypes (A) UMAP projection of MSN nuclei. Each dot represents a nucleus, and the colors represent the different MSN types. (B) Feature plots for the expression of DRD1, TAC1, DRD2, and PENK showing the separation of D1- and D2-MSNs and the expression of marker genes enriched in each cluster. (C) Heatmap showing the top ten most enriched genes in each MSN type. Colored bar at the top corresponds to the colors in (A). (D) Top: MSN-type identifications colored according to (A). Bottom: violin plots showing cell-type- and compartment-specific marker gene expression. (E) The accuracy rate between SCCAF-decoded cell type and actual cell type using the data combined from both subjects. See also Figures S1 and S3, Table S1, and Data S1.

Article Snippet: To verify DRD2 and CPNE4 labels cholinergic neurons, after FISH labeling with the two probes, we performed the following immunostaining with ChAT antibodies (ProSci, Cat# 45-037, 1:1000) using similar approach except that TBS buffer instead of PT buffer was used.

Techniques: Expressing, Marker, Gene Expression

Journal: Current biology : CB

Article Title: Transcriptional and anatomical diversity of medium spiny neurons in the primate striatum.

doi: 10.1016/j.cub.2021.10.015

Figure Lengend Snippet: Figure 4. MSN subtypes in the dorsal striatum (A) FISH labeling of DRD1 (green) and DRD2 (magenta). White box indicates the region shown in (B). The top right axis shows D, V, L, and M di- rections. (B) High-resolution image of region highlighted in (A). (C) Schematic diagrams of the three sections used for DRD1 and DRD2 quantification. The square boxes indicate the quantified regions of interest (ROIs). (D) Quantification of cell density of neurons ex- pressing DRD1 (green), DRD2 (magenta), or both (orange) in the Cd and Pt. Error bars are SD across ROIs. (E) One example MSN expressing both DRD1 and DRD2. (F) RXFP1 labels D1/D2-hybrid MSNs in the dorsal striatum. Arrowhead points to an example D1/D2- hybrid cell. (G) Quantification of DRD1 and DRD2 grain number in D1/D2-hybrid cells and normal D1- or D2-MSNs. Unpaired t test was used for statistical analysis, and p values were indicated on the plots. Error bars represent standard deviation (SD) across 6 cells per type. NS, non-specific. (H) FISH labeling of DRD1 and RXFP1. Left two pictures are original FISH images showing the dis- tribution of DRD1 and RXFP1. The right two pic- tures are CellProfiler-processed images showing DRD1 expressing (red dots) or DRD1 and RXFP1 (black dots, enlarged for display purposes) co-ex- pressing cells. Gray dots are nuclei. (I) Immunohistochemistry of KChIP1 showing robust striosome pattern. (J) FISH labeling of KCNIP1 (yellow) and STXBP6 (blue) distinguishes striosome and matrix, respec- tively. White square indicated the region shown in (K). (K) Top: detail of the white square in (J). Bottom: as above, for characteristic striosome and matrix markers, BACH2 and GDA. Cd, caudate; Pt, putamen; IC, internal capsule. See also Figure S4, Table S1, and Data S4.

Article Snippet: To verify DRD2 and CPNE4 labels cholinergic neurons, after FISH labeling with the two probes, we performed the following immunostaining with ChAT antibodies (ProSci, Cat# 45-037, 1:1000) using similar approach except that TBS buffer instead of PT buffer was used.

Techniques: Labeling, Expressing, Standard Deviation, Immunohistochemistry

Journal: Current biology : CB

Article Title: Transcriptional and anatomical diversity of medium spiny neurons in the primate striatum.

doi: 10.1016/j.cub.2021.10.015

Figure Lengend Snippet: Figure 5. MSN subtypes in the VS (A) Top: double labeling with DRD1 (left) and ARHGAP6 (middle) of the NAs shell/OT shows that ARHGAP6 is selectively enriched in the shell/OT. Bottom: CellProfiler-processed images for the above images. Lettered boxes indicate regions shown in Figure S5A and S5B. (B) Left: quantification of grain number of ARHGAP6 in shell/OT and core. Unpaired t test was used for statistical analysis. Error bars represent SD across thirty- two cells per each region. Right: quantification of grain number of GREB1L in shell/OT and core. Unpaired t test was used for statistical analysis. Error bars represent SD across twenty-nine cells per each region. (C) Violin plot showing TAC3 levels in D1Sh-TAC3 archetype, other D1Sh cells, and TAC3 interneurons. (D) TAC3 co-localizes with DRD1 in medial shell MSNs. (E) DRD2 FISH image showing the outline of the striatum. Dashed white line delineate the borders of striatum. (F) CellProfiler results showing theTAC3 distribution (red dots) in the section in (E). Gray dots are nuclei. (G) CellProfiler results showing the distribution of TAC3 and DRD1 co-expressing cells (dark green dots) in the section in (E). (H) CellProfiler results showing the distribution of TAC3 and DRD2 co-expressing cells (dark green dots) in the section in (E). (I) CellProfiler results showing the distribution of TAC3 and DRD1 co-expressing cells in a second animal (dark green dots). See also Figure S5, Tables S1 and S3, and Data S3 and S4.

Article Snippet: To verify DRD2 and CPNE4 labels cholinergic neurons, after FISH labeling with the two probes, we performed the following immunostaining with ChAT antibodies (ProSci, Cat# 45-037, 1:1000) using similar approach except that TBS buffer instead of PT buffer was used.

Techniques: Labeling, Expressing

Journal: Current biology : CB

Article Title: Transcriptional and anatomical diversity of medium spiny neurons in the primate striatum.

doi: 10.1016/j.cub.2021.10.015

Figure Lengend Snippet: Figure 6. Cell types in the interface islands (A) FISH stain of DRD1 (green) and DRD2 (magenta). Inset: white area indicates striatum, and the red box highlights the area shown in (A) and (B). (B) FISH stain of DRD1, RXFP1, and CPNE4 in immediately adjacent section from (A). (C) High-resolution image of the regions indicated with the letter ‘‘C’’ in (B). (D) High-resolution image of the region indicated with the letter ‘‘D’’ in (B). (E) Distribution of RXFP1 and CPNE4 clusters across six rostral-caudal (R-C) regions identified by multichannel FISH. The upper right axis shows D, V, L, M, R, and C directions. Dashed white box denotes RXFP1 clusters in Pt, the FISH image of which is shown in Figure S5H. Yellow arrowheads point to RXFP1 clusters in caudal extent of NAc in both illustration and images shown in (F). (F) Example RXFP1 clusters in caudal extent of NAc. AC, anterior commissure; GPe, external globus pallidus; VP, ventral pallidum. See also Figures S5–S7, Table S1, and Data S4.

Article Snippet: To verify DRD2 and CPNE4 labels cholinergic neurons, after FISH labeling with the two probes, we performed the following immunostaining with ChAT antibodies (ProSci, Cat# 45-037, 1:1000) using similar approach except that TBS buffer instead of PT buffer was used.

Techniques: Staining