ventricle  (MedChemExpress)

Journal: bioRxiv

Article Title: P2X7 receptor-mediated astrocytic atrophy in the hippocampus of mice after status epilepticus

doi: 10.64898/2026.04.16.718853

Figure Lengend Snippet: Experimental hypothesis and workflow for investigating astrocytic P2X7R receptor signaling in kainic acid–induced epilepsy. Hypothesis: Epileptic seizures trigger ATP release and P2X7 receptor activation, leading to morphological atrophy and functional impairment of hippocampal astrocytes. Workflow: Adult C57BL/6 mice received into the hippocampus viral injections for astrocyte-specific labeling with mCherry. After 3 weeks, epilepsy was induced by intraperitoneal (i.p.) injection of kainic acid (KA). For morphological analyses, brains were collected 2 weeks after KA administration, followed by immunofluorescence staining, image acquisition, and 3D reconstruction of astrocytes. In parallel, for electrophysiological recordings, acute hippocampal slices were prepared at 1 h or 2 weeks after KA treatment and subjected to patch-clamp recordings to assess astrocytic functional properties.

Article Snippet: The drugs used were the following: JNJ-47965567, gabazine hydrobromide (Tocris Biosciences, Bristol, UK); kainic acid hydrate (MedChem Express, Monmouth Junction, NJ, USA); diazepam (Shanghai Xudong Haipu Pharmaceutical Co., Shanghai, China).

Techniques: Activation Assay, Functional Assay, Labeling, Injection, Immunofluorescence, Staining, Patch Clamp

Journal: bioRxiv

Article Title: P2X7 receptor-mediated astrocytic atrophy in the hippocampus of mice after status epilepticus

doi: 10.64898/2026.04.16.718853

Figure Lengend Snippet: JNJ-47965567 and diazepam-treatment preserve astrocytic morphology after kainic acid-induced atrophy in the hippocampal stratum oriens . All abbreviations in this and the - were as defined in . (A) Representative 3D surface reconstructions of hippocampal mCherry-labeled astrocytes (red) from mice treated with NS or kainic acid KA. Scale bar, 5 μm. (B, C) Morphometric quantification of astrocytes showing (B) astrocytic volume (**P<0.01) and (C) primary branch length (***P<0.001). (D) Schematic representation of Sholl analysis used to assess astrocytic complexity. (E) Maximal number of intersections (***P<0.001). (B-E) The unpaired two-tailed t test was used for statistical evaluation. (F) Intersection profile relative to the distance from the soma. (G) Representative 3D surface reconstructions of hippocampal mCherry-labeled astrocytes (red) from KA-, JNJ+KA-, and DZP+KA-treated mice. Scale bar, 5 μm. (H, I) Morphometric quantification of astrocytes showing (H) astrocytic volume (*P<0.05; **P<0.01) and (I) primary branch length (*P<0.05, **P<0.01). (J) Schematic representation of Sholl analysis. (K) Maximal number of intersections (*P<0.05, ***P<0.001) (H-I, K) One-way ANOVA followed by the Tukey’s test was used for statistical evaluaton. (L) Intersection profile relative to the distance from the soma. All data are presented as mean±S.E.M. Dots represent individual astrocytic values (n=15).

Article Snippet: The drugs used were the following: JNJ-47965567, gabazine hydrobromide (Tocris Biosciences, Bristol, UK); kainic acid hydrate (MedChem Express, Monmouth Junction, NJ, USA); diazepam (Shanghai Xudong Haipu Pharmaceutical Co., Shanghai, China).

Techniques: Labeling, Two Tailed Test

Journal: bioRxiv

Article Title: P2X7 receptor-mediated astrocytic atrophy in the hippocampus of mice after status epilepticus

doi: 10.64898/2026.04.16.718853

Figure Lengend Snippet: JNJ-47965567 and diazepam-treatment prevent the reduced expression of ezrin at astrocytic peripheral processes following kainic acid administration in the hippocampal stratum oriens . (A) Representative 3D surface reconstructions of hippocampal mCherry-labeled astrocytes (red) immunolabeled for ezrin (green) from mice treated with NS or KA. Scale bar, 5 μm. (B, C) Quantitative analysis of ezrin colocalization with astrocytes showing (B) ezrin puncta area localized to the astrocytic soma (P>0.05) and (C) ezrin puncta area localized to the astrocytic peripheral branches (***P<0.001). (D) Representative confocal images showing ezrin (green) and mCherry-labeled astrocytes (red); DAPI-stained nuclei (blue) in NS and KA groups. Scale bar, 10 μm. (E) Ezrin fluorescence intensity within the mCherry-defined astrocytic area (***P<0.001) (B, C, E) The Mann-Whitney test was used for statistical evaluation. (F) Representative 3D surface reconstructions of hippocampal mCherry-labeled astrocytes (red) immunolabeled for ezrin (green) from KA-, JNJ+KA-, and DZP+KA-treated mice. Scale bar, 5 μm. (G, H) Quantitative analysis of ezrin colocalization with astrocytes showing (G) ezrin puncta area at the soma (P>0.05) and (H) ezrin puncta area at the branches (**P<0.01, ***P<0.001). (I) Representative confocal images showing ezrin (green) and mCherry-labeled astrocytes (red); DAPI-stained nuclei (blue) in the KA, JNJ+KA, and DZP+KA groups. Scale bar, 10 μm. (J) Ezrin fluorescence intensity within the astrocytic area (**P<0.01) (G, H, J) The Kruskal-Wallis ANOVA followed by the Dunn’s test was used for statistical evaluation. All data are presented as mean±S.E.M. Dots represent individual astrocytic values (n=15).

Article Snippet: The drugs used were the following: JNJ-47965567, gabazine hydrobromide (Tocris Biosciences, Bristol, UK); kainic acid hydrate (MedChem Express, Monmouth Junction, NJ, USA); diazepam (Shanghai Xudong Haipu Pharmaceutical Co., Shanghai, China).

Techniques: Expressing, Labeling, Immunolabeling, Staining, Fluorescence, MANN-WHITNEY

Journal: bioRxiv

Article Title: P2X7 receptor-mediated astrocytic atrophy in the hippocampus of mice after status epilepticus

doi: 10.64898/2026.04.16.718853

Figure Lengend Snippet: JNJ-47965567-treatment attenuates kainic acid-induced reactive astrogliosis in the hippocampal stratum radiatum , whereas there is no effect in the stratum oriens . (A) Representative immunofluorescence images of GFAP- (red) and DAPI-staining (blue) in the hippocampus from mice treated NS, KA, JNJ, or JNJ+KA. Dashed yellow lines delineate the stratum oriens (SO) and stratum radiatum (SR). Scale bar, 200 μm. (B, C) Quantitative analysis of GFAP-positive astrocytic density showing (B) in the SO (P>0.05) and (C) in the SR (**P<0.01, ***P<0.001; in both cases one-way ANOVA followed by the Tukey’s test). (D) Representative immunofluorescence images of S100β- (green) and DAPI-staining (blue) in the hippocampus from mice under the indicated treatment conditions. Dashed yellow lines indicate SO and SR. Scale bar, 200 μm. (E, F) Quantitative analysis of S100β-positive astrocytic density (E) in the SO (P>0.05) and (F) in the SR (P>0.05; in both cases one-way ANOVA followed by the Tukey’s test). All data are presented as mean±S.E.M. Dots represent individual values in 6 mice.

Article Snippet: The drugs used were the following: JNJ-47965567, gabazine hydrobromide (Tocris Biosciences, Bristol, UK); kainic acid hydrate (MedChem Express, Monmouth Junction, NJ, USA); diazepam (Shanghai Xudong Haipu Pharmaceutical Co., Shanghai, China).

Techniques: Immunofluorescence, Staining

Journal: bioRxiv

Article Title: P2X7 receptor-mediated astrocytic atrophy in the hippocampus of mice after status epilepticus

doi: 10.64898/2026.04.16.718853

Figure Lengend Snippet: NMDA, Bz-ATP and muscimol-induced current responses in hippocampal stratum oriens astrocytes of mice. Each agonist was superfused twice for 10 s each with an inter-application interval of 3 min, at a holding potential of −80 mV. The concentrations of the agonists were 100 µM (NMDA, muscimol) and 1000 µM (Bz-ATP) in a low X 2+ bath medium (see Methods). The current response was calculated for each agonist as a mean of the two subsequent applications. Mice were 1 h or 2 weeks before preparation of hippocampal brain slices injected with normal saline (NS) or kainic acid (KA, 30 mg/kg, i.p.). (A) Representative recording 1 h after saline injection. (B) Representative recording 1 h after KA injection. (C) Comparison of the effects of NMDA, Bz-ATP, and muscimol, 1 h after saline or KA-application. P>0.05 all, n=6; Kruskal-Wallis ANOVA, followed by the Dunn’s test. (D) Representative recording 1 h after saline injection in the continuous presence of A438079 (10 µM). (E) Representative recording 1 h after KA injection in the continuous presence of A438079 (10 µM). (F) Comparison of the effects of NMDA, Bz-ATP, and muscimol, 1 h after saline or KA-application in the continuous presence of A438079 (10 µM). P>0.05 all, n=6; one-way ANOVA followed by the Tukey’s test. (G) Comparison of the effects of NMDA, Bz-ATP, and muscimol, 1 h after saline or KA-application. *P<0.05, as indicated; n=6; Kruskal-Wallis ANOVA, followed by the Dunn’s test. (H) Comparison of the effects of NMDA, Bz-ATP, and muscimol, 1 h after saline or KA-application in the continuous presence of A438079 (10 µM). P>0.05 all, n=6; one-way ANOVA followed by the Tukey’s test. For further details see the Materials and Methods Section. All data are presented as mean±S.E.M. Dots represent individual values in 6 mice.

Article Snippet: The drugs used were the following: JNJ-47965567, gabazine hydrobromide (Tocris Biosciences, Bristol, UK); kainic acid hydrate (MedChem Express, Monmouth Junction, NJ, USA); diazepam (Shanghai Xudong Haipu Pharmaceutical Co., Shanghai, China).

Techniques: Injection, Saline, Comparison