Journal: Pain Reports

Article Title: Upregulated spinal histone deacetylases induce nociceptive sensitization by inhibiting the GABA system in chronic constriction injury–induced neuropathy in rats

doi: 10.1097/PR9.0000000000001209

Figure Lengend Snippet: Primary antibodies for western blot and immunofluorescence staining.

Article Snippet: , Anti-CD11b (OX42) , Mouse , CBL1512 , EMD Millipore , 1:500.

Techniques: Western Blot, Immunofluorescence, Staining

Journal: Frontiers in Molecular Neuroscience

Article Title: C286, an orally available retinoic acid receptor β agonist drug, regulates multiple pathways to achieve spinal cord injury repair

doi: 10.3389/fnmol.2024.1411384

Figure Lengend Snippet: C286 induces functional recovery and tissue sparing in a model of spinal cord contusion. (A) C286 treatment leads to significant improvements in locomotor function in hind limbs in comparison to vehicle-treated rats. Data represent the mean ± SEM of n = 4, * p ≤ 0.01, ** p ≤ 0.005. “____” for the statistical significance of the right hind limbs and “___” for the left. One-way Analysis of Variance (ANOVA), followed by the Fisher test. (B) A schematic representation of the spinal cord area used for analysis of the injury pathology and cavity size. (C) Quantification of cavity area (expressed as a percentage of SC area) at 0 and 1 mm caudal to the epicentre at week 6 post-injury. Data represent the mean ± SEM, ** p ≤ 0.005, *** p ≤ 0.001, and Student's t -test. (D–I) C286 treatment leads to improved injury pathology and neuroprotection following a spinal contusion. Quantification of averaged (E) OX42, (F) NeuN, (H) GFAP, and (I) Iba1 expression shown as mean pixels in arbitrary units (a.u.) measured over a series of sections from 0–1 mm caudal to the epicentre revealed a significant preservation of spinal neurons and decreased reactive glia following C286 treatment compared with vehicle treatment. Data represent the mean FI ± SEM, ** p ≤ 0.005, *** p ≤ 0.001, and Student's t -test.

Article Snippet: They were incubated in the secondary antibody for 1 h at room temperature (RT) in PBS-0.02% Tween and then washed in PBS three times for 5 min. Antibodies used were: mouse monoclonal anti-βIII tubulin (Promega, 1:1,000 for immunohistochemistry), chicken polyclonal anti-GFAP (Abcam, 1:300); rabbit polyclonal anti-RARβ (Santa Cruz Biotechnology, Inc, 1:100 for immunohistochemistry), goat polyclonal anti-CGRP (Abcam, 1:200); mouse monoclonal anti-GAP-43 (Millipore, 1:20); mouse monoclonal anti-NF160 (Sigma Aldrich, 1:200); mouse monoclonal anti-OX42 (Abcam, 1:500); rabbit polyclonal anti-Iba1 (DAKO, 1:1,000); mouse monoclonal anti-CSPG (CS-56) (Sigma, 1:100), goat polyclonal anti-decorin (R&D Systems, 1:20, for immunochemistry and 5μg/ml as a neutralising antibody), rabbit polyclonal anti-decorin (Abcam, 1:50), mouse monoclonal anti-Osteopontin Invitrogen (ThermoFisher, 1:250); mouse monoclonal anti- integrin α9 (Abcam, 1:250); mouse monoclonal anti-tenascin-C (Abcam, 1:300).

Techniques: Functional Assay, Comparison, Expressing, Preserving

Journal: International Journal of Molecular Sciences

Article Title: Dynamics of Cellular Regulation of Fractalkine/CX3CL1 and Its Receptor CX3CR1 in the Rat Trigeminal Subnucleus Caudalis after Unilateral Infraorbital Nerve Lesion—Extended Cellular Signaling of the CX3CL1/CX3CR1 Axis in the Development of Trigeminal Neuropathic Pain

doi: 10.3390/ijms25116069

Figure Lengend Snippet: Double immunostaining with anti-CX3CL1 and anti-NeuN antibodies revealed the presence of CX3CL1 in neurons of the superficial layer of the ipsilateral TSC in IONL-operated rats at POD1 ( A – C , arrowheads). To identify glia-like cells immunopositive for CX3CL1, the sections were double immunostained with anti-CX3CL1 and anti-OX42 or anti-GFAP antibodies. The merged image does not demonstrate CX3CL1 immunofluorescence in OX42 immunopositive microglial cells at POD7 (insets in D – F ). The CX3CL1 immunofluorescence was detected in GFAP immunopositive astrocytes of the ipsilateral TSC at POD7 ( G – I , arrows, and insets). The cell nuclei in the merged images are stained blue with Hoechst 33342. Scale bars = 50 µm.

Article Snippet: OX42 , pAb , Chicken , MyBioSource , 1:500 , 18 h.

Techniques: Double Immunostaining, Immunofluorescence, Staining

Journal: International Journal of Molecular Sciences

Article Title: Dynamics of Cellular Regulation of Fractalkine/CX3CL1 and Its Receptor CX3CR1 in the Rat Trigeminal Subnucleus Caudalis after Unilateral Infraorbital Nerve Lesion—Extended Cellular Signaling of the CX3CL1/CX3CR1 Axis in the Development of Trigeminal Neuropathic Pain

doi: 10.3390/ijms25116069

Figure Lengend Snippet: Double immunostaining with anti-CX3CR1 and anti-NeuN antibodies revealed the presence of CX3CR1 in neurons of the superficial layer of the TSC ipsilateral to IONL on POD1 (arrowheads in A – C ). The CX3CR1 immunostaining in microglial cells of the TSC ipsilateral to IONL at POD3 is illustrated in the section double immunostained with anti-CX3CR1 and OX42 antibodies (arrows and insets in D – F ). The double immunostained section through TSC ipsilateral to IONL on POD7 illustrates dominant CX3CR1 immunostaining in GFAP immunopositive astrocytes (arrows and insets in G – I ). The cell nuclei in the merged images are stained blue with Hoechst 33342 staining. Scale bars = 50 µm.

Article Snippet: OX42 , pAb , Chicken , MyBioSource , 1:500 , 18 h.

Techniques: Double Immunostaining, Immunostaining, Staining

Journal: International Journal of Molecular Sciences

Article Title: Dynamics of Cellular Regulation of Fractalkine/CX3CL1 and Its Receptor CX3CR1 in the Rat Trigeminal Subnucleus Caudalis after Unilateral Infraorbital Nerve Lesion—Extended Cellular Signaling of the CX3CL1/CX3CR1 Axis in the Development of Trigeminal Neuropathic Pain

doi: 10.3390/ijms25116069

Figure Lengend Snippet: Panel of representative images illustrating the cellular distribution of Cathepsin-S (CatS) protein, one of two enzymes that are involved in releasing the soluble form of CX3CL1. Double immunostaining with antibodies against CatS and NeuN demonstrates the neuronal distribution of this enzyme protein in TSC ipsilateral to IONL at POD1 ( A – C , arrowheads). The dashed line indicates the boundary between the superficial and deep layers of TSC, indicating that CatS is present only in the neurons of the superficial layer of TSC. The distribution of neuronal CatS was observed bilaterally in TSC at all periods of survival. CatS immunostaining was colocalized with OX42 immunofluorescence, a marker of activated microglial cells, in TSC ipsilateral to IONL at POD3. The arrows indicate immunopositive microglial cells, while the arrowheads indicate CatS immunopositivity in neurons ( D – F , and insets). In contrast, the sections of TSC ipsilateral to IONL at POD7 and POD14 displayed CatS immunofluorescence predominantly colocalized with GFAP immunopositivity, indicating the presence of the enzyme protein in reactive astrocytes. A representative section of the ipsilateral TSC from a rat at POD7 after double immunostaining for CatS and GFAP is illustrated in ( G – I ) and their insets. Scale bars = 50 µm.

Article Snippet: OX42 , pAb , Chicken , MyBioSource , 1:500 , 18 h.

Techniques: Double Immunostaining, Immunostaining, Immunofluorescence, Marker

Journal: International Journal of Molecular Sciences

Article Title: Dynamics of Cellular Regulation of Fractalkine/CX3CL1 and Its Receptor CX3CR1 in the Rat Trigeminal Subnucleus Caudalis after Unilateral Infraorbital Nerve Lesion—Extended Cellular Signaling of the CX3CL1/CX3CR1 Axis in the Development of Trigeminal Neuropathic Pain

doi: 10.3390/ijms25116069

Figure Lengend Snippet: The panel of representative images illustrating the cellular distribution of the ADAM17 protein, which is another enzyme involved in releasing the soluble form of CX3CL1. Unlike CatS, ADAM17 immunopositive neurons were observed in both the superficial and deep layers of TSC, as illustrated by double immunostaining of ADAM17 with NeuN ( A – C , arrowheads, and insets) as a representative section of the ipsilateral TSC at POD3. The intense ADAM17 immunostaining observed in the TSC neurons was bilateral at all time points. The presence of this enzyme protein in activated microglial cells of the ipsilateral TSC was revealed by double immunostaining with anti-ADAM17 and OX42 antibodies, mainly at POD1 and POD3 (arrows and insets in D – F as a representative illustration). The absence of ADAM17 immunofluorescence was observed after double immunostaining with GFAP, indicating that reactive astrocytes did not express this enzyme protein during POD1 and POD3 (arrows and insets in G – I as a representative illustration). However, on POD7 and POD14, ADAM17 immunostaining was found to colocalize with GFAP immunopositive reactive astrocytes. The results of the latter period are used for illustration (arrows and insets in J – L as a representative illustration). Scale bars = 50 µm.

Article Snippet: OX42 , pAb , Chicken , MyBioSource , 1:500 , 18 h.

Techniques: Double Immunostaining, Immunostaining, Immunofluorescence

Journal: International Journal of Molecular Sciences

Article Title: Dynamics of Cellular Regulation of Fractalkine/CX3CL1 and Its Receptor CX3CR1 in the Rat Trigeminal Subnucleus Caudalis after Unilateral Infraorbital Nerve Lesion—Extended Cellular Signaling of the CX3CL1/CX3CR1 Axis in the Development of Trigeminal Neuropathic Pain

doi: 10.3390/ijms25116069

Figure Lengend Snippet: List of primary antibodies used for immunostaining.

Article Snippet: OX42 , pAb , Chicken , MyBioSource , 1:500 , 18 h.

Techniques: Immunostaining

Journal: International Journal of Molecular Sciences

Article Title: Dynamics of Cellular Regulation of Fractalkine/CX3CL1 and Its Receptor CX3CR1 in the Rat Trigeminal Subnucleus Caudalis after Unilateral Infraorbital Nerve Lesion—Extended Cellular Signaling of the CX3CL1/CX3CR1 Axis in the Development of Trigeminal Neuropathic Pain

doi: 10.3390/ijms25116069

Figure Lengend Snippet: Double immunostaining with anti-CX3CL1 and anti-NeuN antibodies revealed the presence of CX3CL1 in neurons of the superficial layer of the ipsilateral TSC in IONL-operated rats at POD1 ( A – C , arrowheads). To identify glia-like cells immunopositive for CX3CL1, the sections were double immunostained with anti-CX3CL1 and anti-OX42 or anti-GFAP antibodies. The merged image does not demonstrate CX3CL1 immunofluorescence in OX42 immunopositive microglial cells at POD7 (insets in D – F ). The CX3CL1 immunofluorescence was detected in GFAP immunopositive astrocytes of the ipsilateral TSC at POD7 ( G – I , arrows, and insets). The cell nuclei in the merged images are stained blue with Hoechst 33342. Scale bars = 50 µm.

Article Snippet: OX42 , mAb , Mouse , Santa Cruz , 1:50 , 18 h.

Techniques: Double Immunostaining, Immunofluorescence, Staining

Journal: International Journal of Molecular Sciences

Article Title: Dynamics of Cellular Regulation of Fractalkine/CX3CL1 and Its Receptor CX3CR1 in the Rat Trigeminal Subnucleus Caudalis after Unilateral Infraorbital Nerve Lesion—Extended Cellular Signaling of the CX3CL1/CX3CR1 Axis in the Development of Trigeminal Neuropathic Pain

doi: 10.3390/ijms25116069

Figure Lengend Snippet: Double immunostaining with anti-CX3CR1 and anti-NeuN antibodies revealed the presence of CX3CR1 in neurons of the superficial layer of the TSC ipsilateral to IONL on POD1 (arrowheads in A – C ). The CX3CR1 immunostaining in microglial cells of the TSC ipsilateral to IONL at POD3 is illustrated in the section double immunostained with anti-CX3CR1 and OX42 antibodies (arrows and insets in D – F ). The double immunostained section through TSC ipsilateral to IONL on POD7 illustrates dominant CX3CR1 immunostaining in GFAP immunopositive astrocytes (arrows and insets in G – I ). The cell nuclei in the merged images are stained blue with Hoechst 33342 staining. Scale bars = 50 µm.

Article Snippet: OX42 , mAb , Mouse , Santa Cruz , 1:50 , 18 h.

Techniques: Double Immunostaining, Immunostaining, Staining

Journal: International Journal of Molecular Sciences

Article Title: Dynamics of Cellular Regulation of Fractalkine/CX3CL1 and Its Receptor CX3CR1 in the Rat Trigeminal Subnucleus Caudalis after Unilateral Infraorbital Nerve Lesion—Extended Cellular Signaling of the CX3CL1/CX3CR1 Axis in the Development of Trigeminal Neuropathic Pain

doi: 10.3390/ijms25116069

Figure Lengend Snippet: Panel of representative images illustrating the cellular distribution of Cathepsin-S (CatS) protein, one of two enzymes that are involved in releasing the soluble form of CX3CL1. Double immunostaining with antibodies against CatS and NeuN demonstrates the neuronal distribution of this enzyme protein in TSC ipsilateral to IONL at POD1 ( A – C , arrowheads). The dashed line indicates the boundary between the superficial and deep layers of TSC, indicating that CatS is present only in the neurons of the superficial layer of TSC. The distribution of neuronal CatS was observed bilaterally in TSC at all periods of survival. CatS immunostaining was colocalized with OX42 immunofluorescence, a marker of activated microglial cells, in TSC ipsilateral to IONL at POD3. The arrows indicate immunopositive microglial cells, while the arrowheads indicate CatS immunopositivity in neurons ( D – F , and insets). In contrast, the sections of TSC ipsilateral to IONL at POD7 and POD14 displayed CatS immunofluorescence predominantly colocalized with GFAP immunopositivity, indicating the presence of the enzyme protein in reactive astrocytes. A representative section of the ipsilateral TSC from a rat at POD7 after double immunostaining for CatS and GFAP is illustrated in ( G – I ) and their insets. Scale bars = 50 µm.

Article Snippet: OX42 , mAb , Mouse , Santa Cruz , 1:50 , 18 h.

Techniques: Double Immunostaining, Immunostaining, Immunofluorescence, Marker

Journal: International Journal of Molecular Sciences

Article Title: Dynamics of Cellular Regulation of Fractalkine/CX3CL1 and Its Receptor CX3CR1 in the Rat Trigeminal Subnucleus Caudalis after Unilateral Infraorbital Nerve Lesion—Extended Cellular Signaling of the CX3CL1/CX3CR1 Axis in the Development of Trigeminal Neuropathic Pain

doi: 10.3390/ijms25116069

Figure Lengend Snippet: The panel of representative images illustrating the cellular distribution of the ADAM17 protein, which is another enzyme involved in releasing the soluble form of CX3CL1. Unlike CatS, ADAM17 immunopositive neurons were observed in both the superficial and deep layers of TSC, as illustrated by double immunostaining of ADAM17 with NeuN ( A – C , arrowheads, and insets) as a representative section of the ipsilateral TSC at POD3. The intense ADAM17 immunostaining observed in the TSC neurons was bilateral at all time points. The presence of this enzyme protein in activated microglial cells of the ipsilateral TSC was revealed by double immunostaining with anti-ADAM17 and OX42 antibodies, mainly at POD1 and POD3 (arrows and insets in D – F as a representative illustration). The absence of ADAM17 immunofluorescence was observed after double immunostaining with GFAP, indicating that reactive astrocytes did not express this enzyme protein during POD1 and POD3 (arrows and insets in G – I as a representative illustration). However, on POD7 and POD14, ADAM17 immunostaining was found to colocalize with GFAP immunopositive reactive astrocytes. The results of the latter period are used for illustration (arrows and insets in J – L as a representative illustration). Scale bars = 50 µm.

Article Snippet: OX42 , mAb , Mouse , Santa Cruz , 1:50 , 18 h.

Techniques: Double Immunostaining, Immunostaining, Immunofluorescence

Journal: International Journal of Molecular Sciences

Article Title: Dynamics of Cellular Regulation of Fractalkine/CX3CL1 and Its Receptor CX3CR1 in the Rat Trigeminal Subnucleus Caudalis after Unilateral Infraorbital Nerve Lesion—Extended Cellular Signaling of the CX3CL1/CX3CR1 Axis in the Development of Trigeminal Neuropathic Pain

doi: 10.3390/ijms25116069

Figure Lengend Snippet: List of primary antibodies used for immunostaining.

Article Snippet: OX42 , mAb , Mouse , Santa Cruz , 1:50 , 18 h.

Techniques: Immunostaining