Journal: Behavioural Pharmacology

Article Title: Naltrexone treatment improves anxiety- and depression-like behavior in alcohol-exposed mice

doi: 10.1097/FBP.0000000000000876

Figure Lengend Snippet: Naltrexone treatment ameliorates microglial activation in alcohol-exposed mice. (a) Immunofluorescence images of microglia labeled with IBA1 (green) in the BLA region (DAPI, blue, for nuclear counterstaining); each image is a representative result from n = 4 independent mice, with images in each treatment group derived from brain tissues of different mice and corresponding coronal sections of the same brain region to ensure intergroup comparability ( n = 4 each group). (b) Quantification of microglial cell numbers in the BLA region ( F = 10.53, DF = 11, P < 0.01; n = 4 each group) * P < 0.05, ** P < 0.01, *** P < 0.001. BLA, basolateral amygdala; DAPI, 4',6-diamidino-2-phenylindole.

Article Snippet: Sections were blocked (5% BSA + 0.5% Triton X-100 in PBS, 1 h, room temperature) and incubated with anti-ionized calcium-binding adapter molecule 1 (IBA1) primary antibody (1 : 500, Proteintech, China, 4°C overnight), followed by Alexa Fluor 488-conjugated secondary antibody (Thermo Fisher Scientific, Waltham, USA, 1 h, room temperature).

Techniques: Activation Assay, Immunofluorescence, Labeling, Derivative Assay

Journal: Biomedicines

Article Title: Sympathetic Regulation of Hematopoiesis and the Mobilization of Inflammatory Cells in ICR Mice with Traumatic Brain Injury: A Novel Approach to Targeting Neuroinflammation and Degenerative Processes

doi: 10.3390/biomedicines13123080

Figure Lengend Snippet: The count of Iba1 + iNOS + cells in the cerebral cortex of male ICR mice at day 7 post-TBI. ( A – C ) Representative micrographs of the cerebral cortex obtained from ( A1 – A3 ) intact mice; ( B1 – B3 ) TBI mice (day 7); and ( C1 – C3 ) TBI mice treated with reserpine. Samples were stained with antibodies specific for ( A1 , B1 , C1 ) Iba1 (red), ( A2 , B2 , C2 ) iNOS (green). DAPI (blue) was used to identify cell nuclei. (Merge) Composite image using all three colors. 4× images. ( D ) Number of Iba1 + cells (% of total DAPI-stained cells); ( E ) number of Iba1 + iNOS + cells and amoeboid Iba1 + cells (% of Iba1 + cells) in the cerebral cortex of male ICR mice. * Significance of difference compared with intact control ( p < 0.017 with the Bonferroni correction). • Significance of difference compared with TBI ( p < 0.017 with the Bonferroni correction). Note: The white circle represents the area of staining for the Iba1 and iNOS markers.

Article Snippet: Specific cellular markers were identified using recombinant mouse monoclonal antibodies against ionized calcium-binding adapter molecule 1 (Iba1) (Cat.No GB15105, Mouse Anti-Iba1, Servicebio Technology, Wuhan, China), rabbit polyclonal antibodies against the inducible nitric oxide synthase (iNOS) (Cat.No GB11119, Rabbit Anti-iNOS, Servicebio Technology, Wuhan, China), rabbit polyclonal antibodies against the macrophage mannose receptor (Mannose Receptor/CD206) (Cat.No GB113497 , Rabbit Anti-Mannose Receptor/CD206, Servicebio Technology, Wuhan, China), rabbit polyclonal antibodies against caspase-3 (Caspase-3) (Cat.No GB11532, Rabbit Anti-Cleaved-Caspase-3, Servicebio Technology, Wuhan, China), rat monoclonal antibody against Myeloperoxidase (MPO) (Cat.No ab300650, Rat Anti-Myeloperoxidase [ EPR20257 ], Abcam, Massachusetts, USA).

Techniques: Staining, Control

Journal: Biomedicines

Article Title: Sympathetic Regulation of Hematopoiesis and the Mobilization of Inflammatory Cells in ICR Mice with Traumatic Brain Injury: A Novel Approach to Targeting Neuroinflammation and Degenerative Processes

doi: 10.3390/biomedicines13123080

Figure Lengend Snippet: The count of Iba1 + iNOS + cells in the subventricular zone of male ICR mice at day 7 post-TBI. ( A – C ) Representative micrographs of the subventricular zone obtained from ( A1 – A3 ) intact mice; ( B1 – B3 ) TBI mice (day 7); and ( C1 – C3 ) TBI mice treated with reserpine. Samples were stained with antibodies specific for ( A1 , B1 , C1 ) Iba1 (red); ( A2 , B2 , C2 ) iNOS (green). DAPI (blue) was used to identify cell nuclei. (Merge) Composite image using all three colors. 4× images. ( D ) Number of Iba1 + cells (% of total cell number); ( E ) number of Iba1 + iNOS + cells and amoeboid Iba1 + cells (% of Iba1 + cells) in the cerebral cortex of male ICR mice. * Significance of difference compared with intact control ( p < 0.017 with the Bonferroni correction). • Significance of difference compared with TBI ( p < 0.017 with the Bonferroni correction). Note: The white circle represents the area of staining for the Iba1 and iNOS markers.

Article Snippet: Specific cellular markers were identified using recombinant mouse monoclonal antibodies against ionized calcium-binding adapter molecule 1 (Iba1) (Cat.No GB15105, Mouse Anti-Iba1, Servicebio Technology, Wuhan, China), rabbit polyclonal antibodies against the inducible nitric oxide synthase (iNOS) (Cat.No GB11119, Rabbit Anti-iNOS, Servicebio Technology, Wuhan, China), rabbit polyclonal antibodies against the macrophage mannose receptor (Mannose Receptor/CD206) (Cat.No GB113497 , Rabbit Anti-Mannose Receptor/CD206, Servicebio Technology, Wuhan, China), rabbit polyclonal antibodies against caspase-3 (Caspase-3) (Cat.No GB11532, Rabbit Anti-Cleaved-Caspase-3, Servicebio Technology, Wuhan, China), rat monoclonal antibody against Myeloperoxidase (MPO) (Cat.No ab300650, Rat Anti-Myeloperoxidase [ EPR20257 ], Abcam, Massachusetts, USA).

Techniques: Staining, Control

Journal: Biomedicines

Article Title: Sympathetic Regulation of Hematopoiesis and the Mobilization of Inflammatory Cells in ICR Mice with Traumatic Brain Injury: A Novel Approach to Targeting Neuroinflammation and Degenerative Processes

doi: 10.3390/biomedicines13123080

Figure Lengend Snippet: The count of Iba1 + Caspase-3 + cells in the cerebral cortex and the subventricular zone of male ICR mice at day 7 post-TBI. Representative micrographs of the ( A – C ) cerebral cortex and ( D – F ) the subventricular zone obtained from ( A , D ) intact mice; ( B – E ) TBI mice (day 7); and ( C – F ) TBI mice treated with reserpine. Samples were stained with antibodies specific for ( A1 , B1 , C1 , D1 , E1 , F1 ) Iba1 (red); ( A2 , B2 , C2 , D2 , E2 , F2 ) Caspase-3 (green). DAPI (blue) was used to identify cell nuclei. ( A3 , B3 , C3 , D3 , E3 , F3 ) (Merge) Composite image using all three colors. 4× images. ( G ) Number of Iba1 + Caspase-3 + cells (% of Iba1 + cells) in the cerebral cortex and subventricular zone of male ICR mice. * Significance of difference compared with intact control ( p < 0.017 with the Bonferroni correction). • Significance of difference compared with TBI ( p < 0.017 with the Bonferroni correction). Note: The white circle represents the area of staining for the Iba1 and Caspase-3 markers.

Article Snippet: Specific cellular markers were identified using recombinant mouse monoclonal antibodies against ionized calcium-binding adapter molecule 1 (Iba1) (Cat.No GB15105, Mouse Anti-Iba1, Servicebio Technology, Wuhan, China), rabbit polyclonal antibodies against the inducible nitric oxide synthase (iNOS) (Cat.No GB11119, Rabbit Anti-iNOS, Servicebio Technology, Wuhan, China), rabbit polyclonal antibodies against the macrophage mannose receptor (Mannose Receptor/CD206) (Cat.No GB113497 , Rabbit Anti-Mannose Receptor/CD206, Servicebio Technology, Wuhan, China), rabbit polyclonal antibodies against caspase-3 (Caspase-3) (Cat.No GB11532, Rabbit Anti-Cleaved-Caspase-3, Servicebio Technology, Wuhan, China), rat monoclonal antibody against Myeloperoxidase (MPO) (Cat.No ab300650, Rat Anti-Myeloperoxidase [ EPR20257 ], Abcam, Massachusetts, USA).

Techniques: Staining, Control

Journal: Biomedicines

Article Title: Sympathetic Regulation of Hematopoiesis and the Mobilization of Inflammatory Cells in ICR Mice with Traumatic Brain Injury: A Novel Approach to Targeting Neuroinflammation and Degenerative Processes

doi: 10.3390/biomedicines13123080

Figure Lengend Snippet: The count of Iba1 + CD206 + cells in the cerebral cortex and subventricular zone of male ICR mice at day 7 post-TBI. Representative micrographs of the ( A – C ) cerebral cortex and ( D – F ) subventricular zone obtained from ( A , D ) intact mice; ( B – E ) TBI mice (day 7); and ( C – F ) TBI mice treated with reserpine. Samples were stained with antibodies specific to ( A1 , B1 , C1 , D1 , E1 , F1 ) Iba1 (red); ( A2 , B2 , C2 , D2 , E2 , F2 ) CD206 + (green). DAPI (blue) was used to identify cell nuclei. ( A3 , B3 , C3 , D3 , E3 , F3 ) (Merge) Composite image using all three colors. 4× images. ( G ) Number of Iba1 + CD206 + cells (% of Iba1 + cells) in the cerebral cortex and subventricular zone of male ICR mice. * Significance of difference compared with intact control ( p < 0.017 with the Bonferroni correction). • Significance of difference compared with TBI ( p < 0.017 with the Bonferroni correction). Note: The white circle represents the area of staining for the Iba1 and CD206 markers.

Article Snippet: Specific cellular markers were identified using recombinant mouse monoclonal antibodies against ionized calcium-binding adapter molecule 1 (Iba1) (Cat.No GB15105, Mouse Anti-Iba1, Servicebio Technology, Wuhan, China), rabbit polyclonal antibodies against the inducible nitric oxide synthase (iNOS) (Cat.No GB11119, Rabbit Anti-iNOS, Servicebio Technology, Wuhan, China), rabbit polyclonal antibodies against the macrophage mannose receptor (Mannose Receptor/CD206) (Cat.No GB113497 , Rabbit Anti-Mannose Receptor/CD206, Servicebio Technology, Wuhan, China), rabbit polyclonal antibodies against caspase-3 (Caspase-3) (Cat.No GB11532, Rabbit Anti-Cleaved-Caspase-3, Servicebio Technology, Wuhan, China), rat monoclonal antibody against Myeloperoxidase (MPO) (Cat.No ab300650, Rat Anti-Myeloperoxidase [ EPR20257 ], Abcam, Massachusetts, USA).

Techniques: Staining, Control

Journal: Biomedicines

Article Title: Sympathetic Regulation of Hematopoiesis and the Mobilization of Inflammatory Cells in ICR Mice with Traumatic Brain Injury: A Novel Approach to Targeting Neuroinflammation and Degenerative Processes

doi: 10.3390/biomedicines13123080

Figure Lengend Snippet: The count of Iba1 + MPO + cells in the cerebral cortex and subventricular zone of male ICR mice at day 7 post-TBI. Representative micrographs of the ( A – C ) cerebral cortex and ( D – F ) subventricular zone obtained from ( A , D ) intact mice; ( B – E ) TBI mice (day 7); and ( C – F ) TBI mice treated with reserpine. Samples were stained with antibodies specific to ( A1 , B1 , C1 , D1 , E1 , F1 ) Iba1 (red); ( A2 , B2 , C2 , D2 , E2 , F2 ) CD206 + (green). DAPI (blue) was used to identify cell nuclei. ( A3,B3,C3,D3,E3,F3 ) (Merge) Composite image using all three colors. 4× images. ( G ) Number of Iba1 + MPO + cells (% of Iba1 + cells) in the cerebral cortex and subventricular zone of male ICR mice. * Significance of difference compared with intact control ( p < 0.017 with the Bonferroni correction). • Significance of difference compared with TBI ( p < 0.017 with the Bonferroni correction). Note: The white circle represents the area of staining for the Iba1 and MPO markers.

Article Snippet: Specific cellular markers were identified using recombinant mouse monoclonal antibodies against ionized calcium-binding adapter molecule 1 (Iba1) (Cat.No GB15105, Mouse Anti-Iba1, Servicebio Technology, Wuhan, China), rabbit polyclonal antibodies against the inducible nitric oxide synthase (iNOS) (Cat.No GB11119, Rabbit Anti-iNOS, Servicebio Technology, Wuhan, China), rabbit polyclonal antibodies against the macrophage mannose receptor (Mannose Receptor/CD206) (Cat.No GB113497 , Rabbit Anti-Mannose Receptor/CD206, Servicebio Technology, Wuhan, China), rabbit polyclonal antibodies against caspase-3 (Caspase-3) (Cat.No GB11532, Rabbit Anti-Cleaved-Caspase-3, Servicebio Technology, Wuhan, China), rat monoclonal antibody against Myeloperoxidase (MPO) (Cat.No ab300650, Rat Anti-Myeloperoxidase [ EPR20257 ], Abcam, Massachusetts, USA).

Techniques: Staining, Control

Journal: Biomedicines

Article Title: Sympathetic Regulation of Hematopoiesis and the Mobilization of Inflammatory Cells in ICR Mice with Traumatic Brain Injury: A Novel Approach to Targeting Neuroinflammation and Degenerative Processes

doi: 10.3390/biomedicines13123080

Figure Lengend Snippet: Possible mechanisms of direct and indirect effects of the sympatholytic drug reserpine on neuroinflammation after TBI. Reserpine exerts its effects via two potential mechanisms. The first mechanism is mediated through the attenuation of the sympathetic nervous system’s (SNS) influence on the bone marrow and immune cells. This occurs via the inhibition of vesicular monoamine transporters 1 and 2 (VMAT 1/2), leading to subsequent degradation of catecholamines in the cytosol by monoamine oxidase (MAO). The resultant decrease in catecholamine concentration in the intercellular space reduces the sympathetic tone on the bone marrow niche and immune cells. This ultimately diminishes the mobilization of immune cells from the bone marrow to the peripheral blood and their migration to the injury site. The second mechanism involves the direct action of reserpine on microglial cells in the brain. Upon penetrating the blood–brain barrier (BBB), reserpine inhibits intracellular soluble epoxide hydrolase (sEH). This inhibition enhances the concentration of epoxyeicosatrienoic acids (EETs), which subsequently suppress the pro-inflammatory NF-κB signaling pathway. This leads to a reduction in inducible nitric oxide synthase (iNOS) production by M1-polarized microglia (Iba1+ cells), thereby decreasing the production of pro-inflammatory cytokines. Concurrently, EETs promote the production of brain-derived neurotrophic factors (BDNFs) and vascular endothelial growth factors (VEGFs) by neurons and astrocytes, enhancing cell survival and regeneration. Collectively, these actions result in reduced neuroinflammation and tissue degeneration.

Article Snippet: Specific cellular markers were identified using recombinant mouse monoclonal antibodies against ionized calcium-binding adapter molecule 1 (Iba1) (Cat.No GB15105, Mouse Anti-Iba1, Servicebio Technology, Wuhan, China), rabbit polyclonal antibodies against the inducible nitric oxide synthase (iNOS) (Cat.No GB11119, Rabbit Anti-iNOS, Servicebio Technology, Wuhan, China), rabbit polyclonal antibodies against the macrophage mannose receptor (Mannose Receptor/CD206) (Cat.No GB113497 , Rabbit Anti-Mannose Receptor/CD206, Servicebio Technology, Wuhan, China), rabbit polyclonal antibodies against caspase-3 (Caspase-3) (Cat.No GB11532, Rabbit Anti-Cleaved-Caspase-3, Servicebio Technology, Wuhan, China), rat monoclonal antibody against Myeloperoxidase (MPO) (Cat.No ab300650, Rat Anti-Myeloperoxidase [ EPR20257 ], Abcam, Massachusetts, USA).

Techniques: Inhibition, Concentration Assay, Migration, Derivative Assay