cx3cr1 gfp ccr2 rfp transgenic mice  (Jackson Laboratory)

Journal: Biomedical Optics Express

Article Title: Histology correlated adaptive optics polarisation sensitive optical coherence tomography

doi: 10.1364/BOE.590238

Figure Lengend Snippet: Top row: log-scale reflectance and DOPU of albino mouse retina. Second row: log-scale reflectance and DOPU of C57BL/6 mouse retina. Third row: log-scale reflectance and DOPU of Cx3cr1 GFP Ccr2 RFP mouse retina. Retinal layers as indicated on reflectance image in top row: retinal nerve fibre layer (RNFL); inner plexiform layer (IPL); inner nuclear layer (INL); outer plexiform layer (OPL); outer nuclear layer (ONL); external limiting membrane (ELM); photoreceptor outer segments (OS); photoreceptor inner segments (IS); retinal pigment epithelium (RPE); choroid.

Article Snippet: Cx3cr1 GFP Ccr2 RFP transgenic mice (Jackson Laboratory, strain no. 032127; RRID: IMSR_JAX:032127) were used to generate time-course images.

Techniques: Membrane

Journal: Biomedical Optics Express

Article Title: Histology correlated adaptive optics polarisation sensitive optical coherence tomography

doi: 10.1364/BOE.590238

Figure Lengend Snippet: Top row: log-scale reflectance and DOPU of albino mouse retina. Second row: log-scale reflectance and DOPU of C57BL/6 mouse retina. Third row: log-scale reflectance and DOPU of Cx3cr1 GFP Ccr2 RFP mouse retina. Retinal layers as indicated on reflectance image in top row: retinal nerve fibre layer (RNFL); inner plexiform layer (IPL); inner nuclear layer (INL); outer plexiform layer (OPL); outer nuclear layer (ONL); external limiting membrane (ELM); photoreceptor outer segments (OS); photoreceptor inner segments (IS); retinal pigment epithelium (RPE); choroid.

Article Snippet: Cx3cr1 GFP Ccr2 RFP mice are derived from a C57BL/6 genetic background.C57BL/6J mice (Charles River, strain no. 027; RRID: IMSR_CRL:027) and albino (Charles River, strain number 022, Crl:CD1(ICR)) mice were used for validation.

Techniques: Membrane

Journal: bioRxiv

Article Title: Meningeal neutrophil infiltration drives inflammation-exacerbated pediatric stroke through IL-36γ signaling

doi: 10.64898/2026.04.03.716365

Figure Lengend Snippet: a, Expression of neutrophil activation markers (ICAM1, iNOS, CD62L and CXCR2) in the ipsilateral hemisphere was assessed by flow cytometry at 24 h post-stroke in PT and LPS/PT groups (N = 7 per group). Data were presented as mean ± S.E.M. ****p < 0.0001 compared with the PT group by unpaired t-test. b-c, Mice were treated with a neutrophil-depleting anti-Ly6G antibody (clone 1A8) or an isotype control antibody beginning one day prior to stroke induction and continuing daily for three consecutive days (P15–P17). Brain sections (1mm sections) were collected 3 days post-stroke and stained with TTC. Infarct volumes were quantified in mm3. (N> 10 per group). *p < 0.05 by one-way ANOVA. d, CCR2–RFP knock-in mice, including homozygous CCR2-deficient (CCR2RFP/RFP) and heterozygous CCR2-sufficient (CCR2RFP/+) littermates, were subjected to PT or LPS/PT. Brain sections were harvested 3 days post-stroke and infarct volumes were quantified by TTC staining and expressed as a percentage of the contralateral hemisphere (N > 5 per group). ns, Not significant, p>0.05; **p < 0.01; ***p < 0.001 by one-way ANOVA.

Article Snippet: C57BL/6J, CCR2 RFP/RFP (JAX#017586), and GFAP-Cre (JAX#012886) mice were purchased from The Jackson Laboratory.

Techniques: Expressing, Activation Assay, Flow Cytometry, Control, Staining, Knock-In

Journal: Scientific Reports

Article Title: Macrophages recruited by implanted fibrin gels promote regeneration of injured lymphatic vessels

doi: 10.1038/s41598-026-39167-2

Figure Lengend Snippet: Fibrin gel implantation following lymphatic injury results in macrophage and neutrophil recruitment. ( a ) Percentage of macrophage infiltration on day 10 after lymphatic injury in the absence and presence of fibrin gel measured by flow cytometry. ( b ) Representative confocal images of the tissue surrounding uninjured and injured collecting lymphatic vessels with gel implantation. The data show that two weeks post-gel implantation, CX3CR1(+) and CCR2(+) cells accumulate around the regenerating vessel, where lectin-647 (white) indicates saphenous lymphatic vessels (see also Supplementary Fig. ). ( c , d ) The top panel illustrates the percentage of various cell types within the CD45 + population. The bottom panel shows the percentage of CCR5 + cells within each identified population. Each group comprised 4 mice, with tissue dissection and flow cytometry conducted 10 days following injury and gel implantation. ANOVA and post-hoc Tukey HSD tests were utilized to determine statistical significance. P-values are presented for statistically significant groups.

Article Snippet: PROX1-eGFP (kind gift of Taija Mäkinen, Uppsala University, and Young-Kwon Hong, University of Southern California), CX3CR1 GFP/+ and CX3CR1 GFP/+ CCR2 RFP/+ reporter mice (C57BL/6, JAX), double mutant CX3CR1 GFP/RFP CCR2 RFP/RFP mice (C57BL/6, JAX), and CCL5-KO mice (C57BL/6, JAX) were housed in MGH Center for Comparative Medicine facilities.

Techniques: Flow Cytometry, Dissection

Journal: Scientific Reports

Article Title: Macrophages recruited by implanted fibrin gels promote regeneration of injured lymphatic vessels

doi: 10.1038/s41598-026-39167-2

Figure Lengend Snippet: Macrophage and neutrophil depletion abolishes lymphatic regeneration after fibrin gel implantation. ( a ) Representative epifluorescence and confocal images of mouse saphenous tissue treated with control and clodronate liposomes after injury showing clodronate liposomes abolished lymphatic endothelial sprouting (PROX1-GFP+) within the fibrin gel. Mice treated with control liposomes exhibit lymphatic sprouting and reconnection within the gel. The fibrin gel was loaded with 10% v/v clodronate or control liposomes prior to polymerization in vivo. The white circle indicates the approximate injury site, and the white arrows show the direction of lymphatic flow. ( b ) Longitudinal near-infrared imaging was used to assess lymphatic pumping and regeneration after macrophage depletion by clodronate liposomes ( n = 4) and control liposomes ( n = 4) using CX3CR1GFP/+ mice. Representative NIR images of the popliteal collecting lymphatic vessels before surgery, two and four weeks after fibrin gel implantation. The red and blue arrows indicate lymphatics distal and proximal to the injury site, respectively. ( c ) Analysis of functional NIR imaging reveals a lack of lymphatic regeneration and loss of lymphatic pumping (packet frequency) after treatment by clodronate liposomes proximal to the injury site. ( d ) After gel implantation and treatment with clodronate liposomes in CX3CR1 GFP/+ mice, injured collecting lymphatic vessels did not adequately regenerate to re-connect the injured vessels (failure of proximal filling). In contrast, CX3CR1 GFP/+ mice treated with control liposomes successfully reconnected injured vessels (success of proximal filling two-sided Fisher’s exact test p < 0.05). ( e ) Confocal imaging was used to evaluate the depletion success in CX3CR1 GFP/+ mice. Quantification of CX3CR1 + cells shows that the number of cells significantly decreased following treatment by clodronate liposomes ( n = 3). Statistically significant differences between presurgical values and values on days 14, and 28 following injury were tested via ANOVA with Tukey’s post hoc tests (* p < 0.05, ** p < 0.01). An unpaired t-test was used to test for significant differences between the control liposomes and clodronate liposome groups at each time (* p < 0.05, ** p < 0.01).

Article Snippet: PROX1-eGFP (kind gift of Taija Mäkinen, Uppsala University, and Young-Kwon Hong, University of Southern California), CX3CR1 GFP/+ and CX3CR1 GFP/+ CCR2 RFP/+ reporter mice (C57BL/6, JAX), double mutant CX3CR1 GFP/RFP CCR2 RFP/RFP mice (C57BL/6, JAX), and CCL5-KO mice (C57BL/6, JAX) were housed in MGH Center for Comparative Medicine facilities.

Techniques: Control, Liposomes, In Vivo, Imaging, Functional Assay

Journal: Scientific Reports

Article Title: Macrophages recruited by implanted fibrin gels promote regeneration of injured lymphatic vessels

doi: 10.1038/s41598-026-39167-2

Figure Lengend Snippet: Fibrin gel implantation leads to increased tissue CCL5 after lymphatic vessel injury and loss of CCL5 inhibits the recovery of lymphatic filling proximal to the injury site. ( a ) A comparative analysis of the levels of cytokines and growth factors after gel implantation demonstrating the upregulation of CCL5 and MMP9 ( n = 3 mice pooled in each group). Data generated by Proteome Profiler Cytokine Array (R&D systems, Minneapolis, MN) according to the manufacturer’s protocol. The values indicate fold changes in different groups relative to the values in the sham group. ( b ) Mice lacking CCL5 (CCL5-/-) cytokine are less likely than control mice to regain filling of vessels proximal to the injury site (proximal filling) subsequent to gel implantation 4-week post-injury ( p = 0.0498, two-sided Fisher’s exact test) while deficiency of CX3CR1 and CCR2 are not associated with the success/failure of proximal filling recovery following gel implantation ( p = 0.5105, two-sided Fisher’s exact test). ( c ) A comparison of lymphatic pumping frequencies via NIR technique proximal and distal to the injury site among CCL5-KO, CX3CR1(GFP/GFP), CCR2(-/-), and double-deficient CX3CR1(GFP/GFP)-CCR2(RFP/RFP) mice following gel implantation. To test the association between categorical variables (success/ failure of proximal filling after gel implantation), the two-sided Fisher’s exact test was used ( p < 0.05). One-way ANOVA analysis with the Tukey post hoc test was used to test the significant difference between different groups after gel implantation (* p < 0.05, ** p < 0.01, *** p < 0.001). All data were presented as mean (± s.e.m.).

Article Snippet: PROX1-eGFP (kind gift of Taija Mäkinen, Uppsala University, and Young-Kwon Hong, University of Southern California), CX3CR1 GFP/+ and CX3CR1 GFP/+ CCR2 RFP/+ reporter mice (C57BL/6, JAX), double mutant CX3CR1 GFP/RFP CCR2 RFP/RFP mice (C57BL/6, JAX), and CCL5-KO mice (C57BL/6, JAX) were housed in MGH Center for Comparative Medicine facilities.

Techniques: Generated, Control, Comparison

Journal: bioRxiv

Article Title: GM-CSF instructs vertebral emergency myelopoiesis in neuropathic pain

doi: 10.1101/2025.11.23.690022

Figure Lengend Snippet: a. SEM image of the inner surface of the L4 vertebral arch. Ossified channels openings (dash circles) on the bone surface (right panel). Scale bars, 500 μm (left) and 100 μm (right). b. Representative maximum intensity projection of the transversal view of the lumbar vertebra (L4) imaged by μ-CT (left panels). High magnification showing ossified channels in the L4 vertebral arch connecting marrow cavity and spinal canal (right) Scale bars, 500 µm (left) and 50 μm (right). c. Digital section in the sagittal plane of a representative human lumbar vertebra imaged by μ-CT. The dashed circles (1 and 2) in the left panel indicate the opening of the ossified channel crossing the cortical bone. Arrows (right panels) show the channels connecting marrow cavity and spinal canal. Scale bars, 2.5 mm. d. Schematic illustration of the protocol to prepare histological specimens from the whole lumbar spinal column. Three consecutive sections (30 μm each) were sliced in the transversal plane and prepared for confocal imaging. e. Representative confocal image showing Ccr2 RFP and Cx3cr1 eGFP positive leukocytes leaving (marked by arrow) the vertebral BM toward subarachnoid space (SAS) through vertebral bone (VB) ossified channel near the L4 DRG. Sample collection was performed 10 days after SNI surgery in Ccr2 RFP - Cx3cr1 eGFP dual reporter mice. Scale bar, 50 μm. f. Scanning electron microscopy image of the outer surface of mouse dorsal dura mater in the DRG meninges. The insertion frame and high magnification (right panel) are showing leukocyte crossing a dura mater membrane pore. Sample collection was performed 10 days after SNI. Scale bar, 10 μm (left) and 1 μm (right). g. Schematic illustration of the experimental protocol workflow. C57BL/6J wild-type (WT) mice were subjected to the SNI model. Seven and nine days later, they received an i.t. injection of AMD3100 (5 μg/site), or vehicle followed by pain behaviors analyses (von Frey test) and isolation of DRG meninges and blood for flow cytometry analyses. h. Absolute number of different cell populations in the DRG meninges from control and SNI samples that receive vehicle or AMD3100 (5 μg/site, i.t.) treatments. n = 8 per group. i. Analyses of mechanical pain hypersensitivity by von Frey tests in control and SNI mice that receive vehicle or AMD3100 (5 μg/site, i.t.) treatments. n = 8 per group. Statistical analysis: one-way ANOVA with Tukey post-tests (h, i). *** P < 0.001, **** P < 0.0001, ns = non-significant. n = biologically independent samples from individual mice or mouse tissues. Error bars indicate the mean ± s.e.m. Illustrations were created with BioRender.com ( https://biorender.com ).

Article Snippet: Ccr2 RFP -Cx3cr1 eGFP dual-reporter mice is a double mutant strain produced by crossing Ccr2 RFP (B6.129(Cg)-Ccr2tm2.1Ifc/J, Jackson Laboratory, Strain: 017586) and Cx3cr1 eGFP (B6.129P-Cx3cr1tm1Litt/J, Jackson Laboratory, Strain: 005582).

Techniques: Imaging, Electron Microscopy, Membrane, Injection, Isolation, Flow Cytometry, Control

Journal: bioRxiv

Article Title: GM-CSF instructs vertebral emergency myelopoiesis in neuropathic pain

doi: 10.1101/2025.11.23.690022

Figure Lengend Snippet: a. Digital section showing antero- lateral view of the lumbar vertebra (L4) imaged by micro-CT. Ossified channels openings (dashed circle) on the inner surface of the vertebral bone. Scale bar, 500 μm. b. Representative confocal image from maximum intensity projection of lumbar vertebra from Ccr2 RFP Cx3cr1 eGFP dual reporter mice. Ccr2 RFP and Cx3cr1 eGFP positive leukocytes in the vertebral BM, inside the vertebra bone (VB) ossified channel near the L4 DRG. The confocal image focused on the vertebral arch region showing an ossified channel traversing the VB to connect the vertebral BM to the subarachnoid space (SAS) near the L4 DRG. Sample collection was performed 10 days after SNI surgery in Ccr2 RFP Cx3cr1 eGFP double transgenic mice. Scale bar, 50 μm. c. Confocal image focused on the contralateral (CL) and ipsilateral (Ipsi) region near the dorsal part of L4 DRG. Arrow indicates accumulated cells. Sample collection was performed 10 days after SNI surgery. Scale bar, 50 μm. d. Flow cytometry gating strategy to identify total leukocytes (CD45 + cells) and myeloid cells (neutrophils and classical monocytes) in the blood of control or SNI mice treated with vehicle or AMD3100. e. Absolute number of leukocytes and myeloid cells (neutrophils and monocytes) in the blood of control or SNI mice treated with vehicle or AMD3100 (5 μg/5 μL, i.t.). n = 8 per group. Statistical analysis: one-way ANOVA with Tukey post-test (e). ns = non-significant. n = biologically independent samples from individual mice or mouse tissues. Error bars indicate the mean ± s.e.m.

Article Snippet: Ccr2 RFP -Cx3cr1 eGFP dual-reporter mice is a double mutant strain produced by crossing Ccr2 RFP (B6.129(Cg)-Ccr2tm2.1Ifc/J, Jackson Laboratory, Strain: 017586) and Cx3cr1 eGFP (B6.129P-Cx3cr1tm1Litt/J, Jackson Laboratory, Strain: 005582).

Techniques: Micro-CT, Transgenic Assay, Flow Cytometry, Control