recombinant rat fractalkine  (R&D Systems)

Journal: Nature Communications

Article Title: Microglia regulate motor neuron plasticity via reciprocal fractalkine and adenosine signaling

doi: 10.1038/s41467-024-54619-x

Figure Lengend Snippet: A Schematic depicting intrapleural injections for retrograde siRNA transport (nontargeting controls, siNTg; rat CX3CL1/fractalkine, siFkn) and CtB to phrenic motor neurons. Representative individual, integrated ( ∫ ) and raw phrenic (Phr) nerve bursts taken during baseline and 90 min post-AIH from siNTg- ( top row ) and siFkn-injected rats ( bottom row ) prior to moderate (mAIH; B ) or severe AIH (sAIH; C ). One-minute averages of phrenic nerve amplitude were measured 90 min post-AIH, and are presented as percent change from baseline ( D ); there was a statistically significant interaction between siRNA (siNTg vs siFkn) and AIH protocol (moderate vs severe) on pLTF (siNTg: n = 4 rats each with 1 independent recording per group; siFkn: n = 7 rats, each with 1 independent recording per group; F(1,18) = 45.431, p < 0.001; two-way ANOVA). * p < 0.01 (siNTg vs siFkn: moderate, p = 0.002; severe, p < 0.001; mAIH vs sAIH: siNTg, p = 0.006; siFkn, p < 0.001; Tukey post-hoc Test). E Representative confocal microscope image of cervical ventral horn; phrenic motor nucleus circled; phrenic motor neurons identified with CtB (green); scale bar (10x magnification): 150 µm; spinal tissue from n = 10 rats. F Representative confocal microscope images from siNTg (left) and siFkn (right)-injected groups stained for CtB (green) and Fkn mRNA (red); scale bar (40x magnification): 20 µm; spinal tissue from n = 5 rats per group. G Fkn mRNA fluorescence intensity was significantly reduced in siFkn vs siNTg phrenic motor neurons (spinal tissue from n = 5 rats per group, at least 10 sections per rat; t(8) = 4.812, p = 0.001; unpaired t-test, two-sided). H Fractalkine mRNA fluorescent intensity in non-phrenic motor neurons was similar between siNTg and siFkn groups (t(8) = 0.542, p = 0.602; unpaired t-test, two-sided). Bars are means ± SEM. Source data are provided as a Source Data file. Figures created in BioRender. Marciante, A. (2024) https://BioRender.com/e55f542 .

Article Snippet: Drugs used include MSX-3 (A2A receptor antagonist; #M3568; Millipore Sigma), Recombinant Rat CX3CL1 protein (#537-FT-025; FisherSci), AZD8797 (CX3CR1 antagonist; #2255; Axon Medchem), JMS-17-2 (selective CX3CR1 antagonist; #HY-123918; MedChemExpress) and ARL67156 (selective ecto-ATPase inhibitor; #128310; FisherSci).

Techniques: Injection, Microscopy, Staining, Fluorescence

Journal: bioRxiv

Article Title: Microglia regulate motor neuron plasticity via reciprocal fractalkine/adenosine signaling

doi: 10.1101/2024.05.07.592939

Figure Lengend Snippet: ( A ) Schematic depicting intrapleural injections for retrograde siRNA transport (nontargeting controls, siNTg; rat CX3CL1/fractalkine, siFkn) and CtB to phrenic motor neurons. ( B , C ) Representative individual, integrated ( ∫ ) and raw phrenic (Phr) nerve bursts taken during baseline and 90 min post-AIH from siNTg-( top row ) and siFkn-injected rats ( bottom row ) prior to moderate (mAIH; B ) or severe AIH (sAIH; C ). One-minute averages of phrenic nerve amplitude were measured 90 min post-AIH, and are presented as percent change from baseline ( D ); there was a statistically significant interaction between siRNA (siNTg versus siFkn) and AIH protocol (moderate vs severe) on pLTF (n=4-7 independent recordings per group; F(1,18) = 45.431, p < 0.001; two-way ANOVA). *p < 0.01; Tukey post-hoc Test. ( E ) Confocal microscope image (10x magnification) of cervical ventral horn; phrenic motor nucleus circled; phrenic motor neurons identified with CtB (green); scale bar (10x magnification): 150 µm. ( F ) Representative confocal microscope images from siNTg ( left ) and siFkn ( right )-injected groups stained for CtB (green) and Fkn mRNA (red); scale bar (40x magnification): 20 µm. ( G ) Fkn mRNA fluorescence intensity was significantly reduced in siFkn vs siNTg phrenic motor neurons (spinal tissue from n=5-6 rats per group with at least 10 sections per rat; t(8) = 4.812, p=0.001; unpaired t -test). ( H ) Fractalkine mRNA fluorescent intensity in non-phrenic motor neurons was similar between siNTg and siFkn groups (t(8) = 0.542, p=0.602; unpaired t-test). Bars are means ± SEM.

Article Snippet: Drugs used include MSX-3 (A 2A receptor antagonist; #M3568; Millipore Sigma), Recombinant Rat CX3CL1 protein (#537-FT-025; FisherSci), AZD8797 (CX3CR1 antagonist; #2255; Axon Medchem), and ARL67156 (selective ecto-ATPase inhibitor; #128310; FisherSci).

Techniques: Injection, Microscopy, Staining, Fluorescence

Journal: Frontiers in Pharmacology

Article Title: CX3CL1 Worsens Cardiorenal Dysfunction and Serves as a Therapeutic Target of Canagliflozin for Cardiorenal Syndrome

doi: 10.3389/fphar.2022.848310

Figure Lengend Snippet: CX3CL1 expression in response to glucose stimulation. (A) Blood glucose levels at 3 and 42 days in streptozotocin (STZ)-induced DM mice. Insulin treatment was initiated at 28 days after STZ injection and persisted for 14 days. * p < 0.05, n = 8 in each group. (B) Blood glucose levels at 3 and 42 days in NOD mice. The NOD mice were assigned to three groups: normal glucose (NG) group (some mice didn’t develop into hyperglycemia), NOD group (with twice random blood glucose level>20 mM) and NOD&insulin group. Insulin treatment protocol was the same as panel (A) . * p < 0.05, n = 8 in each group. (C) Representative western blotting of CX3CL1 in the heart and kidney tissue of mice at various time points after STZ-injection. (D) Semi-quantitation of CX3CL1 expression in panel (C) . * p < 0.05, vs . the corresponding 0 day, n = 5 per group. (E) Immunohistochemical detection of CX3CL1 expression in heart and kidney tissues of normal or diabetic mice. Scale bar = 100 μm. (F) Cardiac CX3CL1 and CX3CR1 mRNA expression in NOD mice. (G) Renal gene expression of CX3CL1 and CX3CR1 in NOD mice. * p < 0.05, vs . NG, n = 6 per group. (H) CX3CL1 expression detected by western blotting in heart and kidney tissues from DM mice with or without insulin treatment for 14 days. (I) Semi-quantitation of CX3CL1 in panel (H) . * p < 0.05 vs . WT&Vehicle, n = 5 in each group. (J) mRNA expression of CX3CL1 in neonatal rat cardiomyocytes (NRCM) and fibroblasts (NRCF), two types of renal cell lines HBZY-1 and NRK-52E exposed to different concentrations of glucose medium for 3 days. * p < 0.05 vs . 5.5 mM group, n = 5 in each group. Experiments presented in panels (A) , (B) , (D) , (I) , and (J) were analyzed using one-way ANOVA followed by Bonferroni’s post hoc test. Experiments presented in panels (F) and (G) were analyzed using two-tailed unpaired t -test.

Article Snippet: Cultured NRCM and HBZY-1 were starved of serum for 12 h and then were exposed to recombinant soluble CX3CL1 (chemokine domain; 537-FT-025; R&D) at 200 ng/ml for 24 h in the presence or absence of 10 μM Y27632 (a ROCK inhibitor; Selleck) or 5 μg/ml of an anti-CX3CR1 antibody (GTX27200; Genetex).

Techniques: Expressing, Injection, Western Blot, Quantitation Assay, Immunohistochemical staining, Gene Expression, Two Tailed Test

Journal: Frontiers in Pharmacology

Article Title: CX3CL1 Worsens Cardiorenal Dysfunction and Serves as a Therapeutic Target of Canagliflozin for Cardiorenal Syndrome

doi: 10.3389/fphar.2022.848310

Figure Lengend Snippet: sCX3CL1 promotes mitochondrial-dependent apoptosis in cardiomyocytes and renal cells. (A) Western blotting to detect RhoA and GTP-bound RhoA protein in neonatal rat cardiomyocytes (NRCM) and HBZY-1 cells after 60 min of stimulation with 200 ng/ml of sCX3CL1 (soluble CX3CL1). (B) Western blotting to detect upregulation of ROCK1 and cleaved ROCK1 (c-ROCK) expression in response to stimulation of NRCM and HBZY-1 cells with sCX3CL1 (200 ng/ml) for 24 h. (C) Semi-quantitation of RhoA-GTP, ROCK and cleaved-ROCK expression. * p < 0.05 vs . control, n = 5 in each group. Representative images of (D) calcein fluorescence or (E) TMRE fluorescence (Mitochondrial membrane potential) in cells cultured with sCX3CL1 alone or co-treated with either a CX3CR1 neutralizing antibody or a Rho kinase inhibitor (Y-27632). Scale bar = 100 μm. Semi-quantitative analysis of calcein (F) or TMRE (G) fluorescence intensity ( * p < 0.05 vs . sCX3CL1), n = 5 in each group. (H) Western blots analysis of Bax, cytochrome C (Cyto C) in NRCM and HBZY-1. Semi-quantitation analysis of Bax and Cyto C in NRCM (I) and HBZY-1 cells (J) . * p < 0.05 vs . sCX3CL1, n = 5 in each group. (K) Subcellular localization of AIF (apoptosis inducing factor) was detected in NRCM and HBZY-1 cells after sCX3CL1 stimulation or co-treatment with either a CX3CR1 neutralizing antibody or Y-27632. Scale bar = 30 μm. Experiments presented in panel (C) was analyzed using two-tailed unpaired t -test and in panels (F) , (G) , (I) , and (J) were analyzed using one-way ANOVA followed by Bonferroni’s post hoc test.

Article Snippet: Cultured NRCM and HBZY-1 were starved of serum for 12 h and then were exposed to recombinant soluble CX3CL1 (chemokine domain; 537-FT-025; R&D) at 200 ng/ml for 24 h in the presence or absence of 10 μM Y27632 (a ROCK inhibitor; Selleck) or 5 μg/ml of an anti-CX3CR1 antibody (GTX27200; Genetex).

Techniques: Western Blot, Expressing, Quantitation Assay, Control, Fluorescence, Membrane, Cell Culture, Two Tailed Test

Journal: Frontiers in Pharmacology

Article Title: CX3CL1 Worsens Cardiorenal Dysfunction and Serves as a Therapeutic Target of Canagliflozin for Cardiorenal Syndrome

doi: 10.3389/fphar.2022.848310

Figure Lengend Snippet: Effect of CX3CL1/CX3CR1 inhibition on cardiorenal dysfunction in mice with diabetes-induced CRS. (A) Representative M-mode echocardiographic images of mice at 6 weeks after STZ injection. Scale bars, 2 mm (upper panels), horizontal bars represent 100 ms. (B) Left ventricular fractional shortening (LVFS). (C) Microalbuminuria amount for 24 h. (D) Serum creatinine levels. (E) Serum NGAL levels. For panels (B–E) , * p < 0.05 vs . WT&STZ group; † p < 0.05 vs . WT&STZ&Insulin group. For panel (B) , n = 6 in WT&Vehicle, WT&STZ&Inuslin, and CX3CR1 −/− &Vehicle group; n = 7 in WT&STZ and CX3CR1 −/− &STZ group; n = 5 in WT&STZ&anti-CX3CL1 group. n = 5 per group in panels (C–E) . (F) Representative M-mode echocardiographic images. Scale bars, 2 mm (upper panels), horizontal bars represent 100 ms. (G) LVFS. (H) Concentrations of serum BUN (urea nitrogen). (I) Serum creatinine levels. (J) Urine creatinine levels. For panel (G–J) , * p < 0.05 vs . NOD&Vehicle group. In panel (G) , n = 8 in NG&Vehicle; n = 6 in NOD&Vehicle, NOD&sh-scr, and NOD&sh-CX3CR1 group; n = 7 in NOD&Inuslin; n = 5 in NOD&Anti-CX3CL1. n = 6 per group in panel (H) ; n = 5 per group in panels (I–J) . Experiments presented in panels (B–E) and (G–J) were analyzed using one-way ANOVA followed by Bonferroni’s post hoc test.

Article Snippet: Cultured NRCM and HBZY-1 were starved of serum for 12 h and then were exposed to recombinant soluble CX3CL1 (chemokine domain; 537-FT-025; R&D) at 200 ng/ml for 24 h in the presence or absence of 10 μM Y27632 (a ROCK inhibitor; Selleck) or 5 μg/ml of an anti-CX3CR1 antibody (GTX27200; Genetex).

Techniques: Inhibition, Injection

Journal: Frontiers in Pharmacology

Article Title: CX3CL1 Worsens Cardiorenal Dysfunction and Serves as a Therapeutic Target of Canagliflozin for Cardiorenal Syndrome

doi: 10.3389/fphar.2022.848310

Figure Lengend Snippet: Inhibition of CX3CL1/CX3CR1 attenuated apoptosis in mice with STZ-induced diabetes. (A) Representative images of TUNEL staining in heart sections at 6 weeks after STZ-injection. (B) Percentage of TUNEL-positive nuclei in heart tissue. (C) Representative images of TUNEL staining in kidney sections at 6 weeks after STZ-injection. (D) Percentage of TUNEL-positive nuclei in kidney tissue. (E) Bax immunostaining (brown staining) in heart tissue. (F) Bax immunostaining in kidney tissue. (G) Representative electronic micrographs of the heart featuring sarcomeres and mitochondria. (H) Representative electronic micrographs of the kidney featuring tubular mitochondria. For panel (B) and (D) , * p < 0.05 vs . WT&STZ mice; † p < 0.05 vs . WT&STZ&Insulin mice, n = 5 in each group. Scale bar = 100 μm in panel (A) and (C) , Scale bar = 1 μm in panel (G) and (H) . Experiments presented in panels (B) and (D) were analyzed using one-way ANOVA followed by Bonferroni’s post hoc test.

Article Snippet: Cultured NRCM and HBZY-1 were starved of serum for 12 h and then were exposed to recombinant soluble CX3CL1 (chemokine domain; 537-FT-025; R&D) at 200 ng/ml for 24 h in the presence or absence of 10 μM Y27632 (a ROCK inhibitor; Selleck) or 5 μg/ml of an anti-CX3CR1 antibody (GTX27200; Genetex).

Techniques: Inhibition, TUNEL Assay, Staining, Injection, Immunostaining, Mouse Assay

Journal: Frontiers in Pharmacology

Article Title: CX3CL1 Worsens Cardiorenal Dysfunction and Serves as a Therapeutic Target of Canagliflozin for Cardiorenal Syndrome

doi: 10.3389/fphar.2022.848310

Figure Lengend Snippet: Inhibition of CX3CL1/CX3CR1 attenuated fibrosis in diabetes mice. (A) Myocardial interstitial fibrosis in STZ-induced DM mice. (B) Perivascular fibrosis of heart in STZ-induced DM mice. (C) Fibrotic score of the renal interstitial area in STZ-induced DM mice. (D) Fibrotic score of the glomeruli area in STZ-induced DM mice. (E) Myocardial interstitial fibrosis in NOD mice. (F) Perivascular fibrosis of heart in NOD mice. (G) Fibrotic score of the renal interstitial area in NOD mice. (H) Fibrotic score of the glomeruli area in NOD mice. (I) Representative images of immunostaining for DAPI (blue), α-SMA (green) and vimentin (red) in heart tissue. Scale bar = 100 μm. (J) Representative images of immunostaining for E-cadherin (red), fibronectin (red), vimentin (red), and α-SMA (green) in kidney tissue. Scale bar = 100 μm. For panels (A–D) , * p < 0.05 vs . WT&STZ group; † p < 0.05 vs . WT&STZ&Insulin mice; n = 5 per group. For panels (E–H) , * p < 0.05 vs . NOD&Vehicle group; n = 6 per group. Experiments presented in panels (A–H) were analyzed using one-way ANOVA followed by Bonferroni’s post hoc test.

Article Snippet: Cultured NRCM and HBZY-1 were starved of serum for 12 h and then were exposed to recombinant soluble CX3CL1 (chemokine domain; 537-FT-025; R&D) at 200 ng/ml for 24 h in the presence or absence of 10 μM Y27632 (a ROCK inhibitor; Selleck) or 5 μg/ml of an anti-CX3CR1 antibody (GTX27200; Genetex).

Techniques: Inhibition, Immunostaining

Journal: Frontiers in Pharmacology

Article Title: CX3CL1 Worsens Cardiorenal Dysfunction and Serves as a Therapeutic Target of Canagliflozin for Cardiorenal Syndrome

doi: 10.3389/fphar.2022.848310

Figure Lengend Snippet: Canagliflozin (Cana) improved cardiorenal dysfunction and repressed diabetic induced or high glucose induced CX3CL1 expression. (A) M-mode of echocardiography. Scale bars, 2 mm (upper panels), horizontal bars represent 100 ms. (B) Left ventricular fractional shortening (LVFS). (C) Serum creatinine concentration. (D) Renal neutrophil gelatinase-associated lipocalin (NGAL) content. (E) CX3CL1 expression detected by Western blotting in heart and kidney tissues. (F) Semi-quantitation of CX3CL1 expression in heart. (G) Semi-quantitation of CX3CL1 expression in kidney. * p < 0.05 vs . WT&STZ mice; n = 6 per group. (H–M) Western blotting of CX3CL1 levels in cultured cardiomyocytes (H9C2), glomerular mesangial cells (HBZY-1), neonatal rat cardiac fibroblasts (NRCF) and renal tubular epithelial cells (NRK-52E). The cultured cells were exposed for 5 days either to normal concentration of glucose (5 mM, NG) or high concentration of glucose (33.3 mM, HG) as well as to HG for 3 days followed by NG for 2 days (HN) with/without treatment of Cana. * p < 0.05 vs . HG; † p < 0.05 vs . HN; n = 5 per group. Experiments presented in panels (B–D) , (F–G) , (I–J) , and (L–M) were analyzed using one-way ANOVA followed by Bonferroni’s post hoc test.

Article Snippet: Cultured NRCM and HBZY-1 were starved of serum for 12 h and then were exposed to recombinant soluble CX3CL1 (chemokine domain; 537-FT-025; R&D) at 200 ng/ml for 24 h in the presence or absence of 10 μM Y27632 (a ROCK inhibitor; Selleck) or 5 μg/ml of an anti-CX3CR1 antibody (GTX27200; Genetex).

Techniques: Expressing, Concentration Assay, Western Blot, Quantitation Assay, Cell Culture

Journal: Frontiers in Pharmacology

Article Title: CX3CL1 Worsens Cardiorenal Dysfunction and Serves as a Therapeutic Target of Canagliflozin for Cardiorenal Syndrome

doi: 10.3389/fphar.2022.848310

Figure Lengend Snippet: Illustration of the molecular mechanisms by which CX3CL1 promotes cardiorenal dysfunction induced by diabetics. CX3CL1 expression is upregulated in cardiac and renal cells by a high glucose environment. Persistent high CX3CL1 expression accelerates the mitochondrial apoptotic pathway through activation of RhoA/ROCK1-Bax signaling. In addition, CX3CL1 regulates fibroblast and epithelial cell phenotypic trans-differentiation through activation of TGF-β/Smad signaling. CX3CL1 leads to the onset of cardiorenal dysfunction in diabetes-induced cardiorenal syndrome type 5 (CRS5) due to its proapoptotic and profibrotic effects, while SGLT2 inhibitor could improve CRS5 at least partially by repressing CX3CL1 expression.

Article Snippet: Cultured NRCM and HBZY-1 were starved of serum for 12 h and then were exposed to recombinant soluble CX3CL1 (chemokine domain; 537-FT-025; R&D) at 200 ng/ml for 24 h in the presence or absence of 10 μM Y27632 (a ROCK inhibitor; Selleck) or 5 μg/ml of an anti-CX3CR1 antibody (GTX27200; Genetex).

Techniques: Expressing, Activation Assay

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Fractalkine-induced microglial vasoregulation occurs within the retina and is altered early in diabetic retinopathy.

doi: 10.1073/pnas.2112561118

Figure Lengend Snippet: Fig. 5. Vasoactive gene expression from retinal microglia and fractalkine-induced vasoconstriction are altered after 4 wk of STZ-induced diabetes. (A) The responsiveness of retinal vessels to hyperoxic challenge was explored in vivo using OCTA (Insets show OCTA images from baseline and after expo- sure to O2). (Scale bar, 200 μm.) While hyperoxic challenge (filled bars) lead to constriction in the control group (n = 10 normoxia, n = 6 100% O2), no constriction was observed in the STZ cohort (n = 12 normoxia, n = 7 100% O2). (B) Microglial vasoregulation was investigated during diabetes, with 4-wk STZ-treated and control retinae exposed to fractalkine ex vivo (representative control and STZ images in Inset) (Scale bar, 50 μm.) While vessels from con- trol retinae showed fractalkine-induced vasoconstriction (filled bar), STZ retinae exhibited no change (n = 5 animals). (C) Differential microglial gene expression data from 4 wk control and STZ-treated animals were compared to vasomodulatory gene lists (vasoconstriction, GO:0097746; angiogenesis, GO:0001525; vasodilation, GO:0097746), with the RAS positive regulator angiotensinogen (Agt), and negative regulator (Ahr) significantly altered (FDR- adjusted, citrate control n = 5, STZ n = 4). (D) OCTA was used to quantify retinal superficial capillary diameter in 4-wk control and STZ-treated animals (unfilled and filled bars, respectively) exposed to candesartan or vehicle. In STZ-treated animals, capillary diameter returned to baseline in the candesartan-treated group (n = 7 control, n = 8, 5 STZ vehicle and candesartan, respectively). (E) Retinal blood flow was quantified using arterio-venous transit time and showed increased transit time (slower blood flow) in STZ-treated animals independent of candesartan treatment (n = 8 control, n = 11 and 8 STZ vehicle and candesartan, respectively). (F) Quantification of the arteriovenous ratio showed candesartan treatment increased the diameter of larger vessels in STZ-treated retinae relative to control and vehicle-treated tissues (n = 8 control, n = 11 and 8 STZ vehicle and candesartan, respectively). Data expressed as mean ± SEM, *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Recombinant rat fractalkine (200 ng/mL; R&D Systems, #537-FT-025/CF) or vehicle (PBS) was introduced after 10 min of baseline recording and imaged for an additional 10 min. At the end of this incubation, vessel diameter was measured at sites with and without microglial contact (2 to 4 individual capillary sites per retina, n = 4 to 6 retinae) and measurements expressed as a percentage of baseline diameter of the same vessel region (taken as the average vessel diameter over the initial 10-min baseline).

Techniques: Gene Expression, In Vivo, Control, Ex Vivo

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Fractalkine-induced microglial vasoregulation occurs within the retina and is altered early in diabetic retinopathy.

doi: 10.1073/pnas.2112561118

Figure Lengend Snippet: Fig. 6. Schematic representation of microglial regulation of ret- inal capillary constriction. Data from this study show microglia are structurally and functionally capable of involvement in the neurovascular unit. Microglia contact neuronal synapses and reti- nal capillaries (including pericytes) and activation of fractalkine- Cx3cr1 signaling results in capillary constriction, which is via an AT1R-dependent mechanism. Ultimately, capillary regulation may occur via direct microglial mechanism or may involve contri- butions from pericytes and M€uller cells. EC, endothelial cell; PC, pericyte.

Article Snippet: Recombinant rat fractalkine (200 ng/mL; R&D Systems, #537-FT-025/CF) or vehicle (PBS) was introduced after 10 min of baseline recording and imaged for an additional 10 min. At the end of this incubation, vessel diameter was measured at sites with and without microglial contact (2 to 4 individual capillary sites per retina, n = 4 to 6 retinae) and measurements expressed as a percentage of baseline diameter of the same vessel region (taken as the average vessel diameter over the initial 10-min baseline).

Techniques: Activation Assay

Journal: Journal of neuropathology and experimental neurology

Article Title: Surgical Trauma Exacerbates Cognitive Deficits and Neuroinflammation in Aged Rats: The Role of CX3CL1-CX3CR1 Signaling.

doi: 10.1093/jnen/nly051

Figure Lengend Snippet: FIGURE 1. Surgical trauma exacerbated neurobehavioral impairment in both adult and aged rats. Age produced an additive effect on center duration in the surgical rats on postoperative day 3. Exogenous CX3CL1 administration rescued these behavioral deficits in the open field test. (A) Total distance moved in the chamber in both adult and aged rats. (B) The time in the central area in both adult and aged rats. The results are represented as the mean 6 SEM. **p < 0.001 versus the day-matched and age-matched control group; #p < 0.05 versus the day-matched and age-matched surgery group; &p < 0.05 versus the day-matched adult surgery group. P3, P7, and P14: postoperative days 3, 7, and 14, respectively. CX3CL1, C-X3-C motif chemokine ligand 1; SEM, standard error of the mean.

Article Snippet: A microsyringe drive (BASi, West Lafayette, IN) operated with a programmable controller (Bee Hive Controller; BASi) delivered recombinant rat CX3CL1 (5 mg in 0.9% NaCl, 5 mL, R&D Systems, Minneapolis, MN) to the animals at a rate of 1 mL/minute in the CX3CL1 group or 0.9% NaCl solution (5 mL) to the animals in the control group.

Techniques: Produced, Control

Journal: Journal of neuropathology and experimental neurology

Article Title: Surgical Trauma Exacerbates Cognitive Deficits and Neuroinflammation in Aged Rats: The Role of CX3CL1-CX3CR1 Signaling.

doi: 10.1093/jnen/nly051

Figure Lengend Snippet: FIGURE 3. Compared with age-matched control group, surgery induced spatial learning and memory impairment in a Morris water maze during a probe trial in both adult and aged rats on postoperative day 3. Treatment with CX3CL1 significantly improved behavioral performance in both adult and aged rats. (A) Cross platform times in both adult and aged rats. (B) Percentage of time spent in target quadrant in both adult and aged rats. (C) Swimming speed in both adult and aged rats. The results are represented as the mean 6 SEM. *p < 0.05, **p < 0.001 versus the day-matched and age- matched control group; #p < 0.05 versus the day-matched and age-matched surgery group. P3, P7, and P14: postoperative days 3, 7, and 14, respectively. CX3CL1, C-X3-C motif chemokine ligand 1; SEM, standard error of the mean.

Article Snippet: A microsyringe drive (BASi, West Lafayette, IN) operated with a programmable controller (Bee Hive Controller; BASi) delivered recombinant rat CX3CL1 (5 mg in 0.9% NaCl, 5 mL, R&D Systems, Minneapolis, MN) to the animals at a rate of 1 mL/minute in the CX3CL1 group or 0.9% NaCl solution (5 mL) to the animals in the control group.

Techniques: Control

Journal: Journal of neuropathology and experimental neurology

Article Title: Surgical Trauma Exacerbates Cognitive Deficits and Neuroinflammation in Aged Rats: The Role of CX3CL1-CX3CR1 Signaling.

doi: 10.1093/jnen/nly051

Figure Lengend Snippet: FIGURE 4. Surgical trauma significantly downregulated CX3CL1 protein expression on postoperative day 3 in the adult rats. However, in the aged rats, the lower expression of CX3CL1 was maintained until day 7 compared with that of the aged controls. The results are represented as the mean 6 SEM. *p < 0.05 versus the age-matched control group; ##p < 0.001 versus the day-matched surgery group. P3, P7, and P14: postoperative days 3, 7, and 14, respectively. C: control; S: surgery; S þ I: surgery þ CX3CL1. CX3CL1, C-X3-C motif chemokine ligand 1; SEM, standard error of the mean.

Article Snippet: A microsyringe drive (BASi, West Lafayette, IN) operated with a programmable controller (Bee Hive Controller; BASi) delivered recombinant rat CX3CL1 (5 mg in 0.9% NaCl, 5 mL, R&D Systems, Minneapolis, MN) to the animals at a rate of 1 mL/minute in the CX3CL1 group or 0.9% NaCl solution (5 mL) to the animals in the control group.

Techniques: Expressing, Control

Journal: Journal of neuropathology and experimental neurology

Article Title: Surgical Trauma Exacerbates Cognitive Deficits and Neuroinflammation in Aged Rats: The Role of CX3CL1-CX3CR1 Signaling.

doi: 10.1093/jnen/nly051

Figure Lengend Snippet: FIGURE 5. The CX3CL1 receptor (CX3CR1) protein was significantly downregulated following surgical trauma in the brains of the adult rats on postoperative day 3. The lower expression of CX3CR1 in the aged rats was maintained until day 7 and returned to baseline on day 14 compared with the age-matched controls. The results are represented as the mean 6 SEM. *p < 0.05, **p < 0.001 versus the day-matched and age-matched control group; #p < 0.05, ##p < 0.001 versus the day-matched and age-matched surgery group. P3, P7, and P14: postoperative days 3, 7, and 14, respectively. C: control; S: surgery; S þ I: surgery þ CX3CL1. CX3CL1, C-X3-C motif chemokine ligand 1; SEM, standard error of the mean.

Article Snippet: A microsyringe drive (BASi, West Lafayette, IN) operated with a programmable controller (Bee Hive Controller; BASi) delivered recombinant rat CX3CL1 (5 mg in 0.9% NaCl, 5 mL, R&D Systems, Minneapolis, MN) to the animals at a rate of 1 mL/minute in the CX3CL1 group or 0.9% NaCl solution (5 mL) to the animals in the control group.

Techniques: Expressing, Control

Journal: Journal of neuropathology and experimental neurology

Article Title: Surgical Trauma Exacerbates Cognitive Deficits and Neuroinflammation in Aged Rats: The Role of CX3CL1-CX3CR1 Signaling.

doi: 10.1093/jnen/nly051

Figure Lengend Snippet: FIGURE 7. There is a significant difference for the basal hippocampal Iba-1 levels between the adult and aged rats. Surgical trauma increased the levels of Iba-1 on postoperative day 3 in the adult rats. However, in the aged rats, the higher levels of Iba-1 were maintained until day 7 compared with that of the aged controls. Treatment with CX3CL1 significantly downregulated the levels of Iba-1 on postoperative days 3, and 7 in the aged rats. (A) Representative images of Iba1- labeled activated microglia in the hippocampus in both adult and aged rats. (B) The IOD of hippocampal CA1 region Iba1- positive cells in both adult and aged rats. The results are represented as the mean 6 SEM. **p < 0.001 versus the day- matched and age-matched control group; ##p < 0.001 versus the day-matched and age-matched surgery group; &&p < 0.001 versus the day-matched adult control group. P3, P7, and P14: postoperative days 3, 7, and 14, respectively. Iba-1, ionized calcium-binding adapter molecule 1; IOD, integrated optical density; CX3CL1, C-X3-C motif chemokine ligand 1; SEM, standard error of the mean. Scale bars: 50 mm.

Article Snippet: A microsyringe drive (BASi, West Lafayette, IN) operated with a programmable controller (Bee Hive Controller; BASi) delivered recombinant rat CX3CL1 (5 mg in 0.9% NaCl, 5 mL, R&D Systems, Minneapolis, MN) to the animals at a rate of 1 mL/minute in the CX3CL1 group or 0.9% NaCl solution (5 mL) to the animals in the control group.

Techniques: Labeling, Control, Binding Assay

Journal: Journal of neuropathology and experimental neurology

Article Title: Surgical Trauma Exacerbates Cognitive Deficits and Neuroinflammation in Aged Rats: The Role of CX3CL1-CX3CR1 Signaling.

doi: 10.1093/jnen/nly051

Figure Lengend Snippet: FIGURE 6. Compared with the day-matched adult surgery group, age amplified and prolonged surgery-induced hippocampal IL-1b protein expression. Treatment with CX3CL1 significantly downregulated the levels of IL-1b in the brains of the aged rats on postoperative days 3 and 7. The similar reduction for IL-1b protein was also observed in the adult rats on postoperative day 3. The results are represented as the mean 6 SEM. *p < 0.05, **p < 0.001 versus the day- matched and age-matched control group; #p < 0.05, ##p < 0.001 versus the day-matched and age-matched surgery group; &&p < 0.001 versus the day-matched adult surgery group. P3, P7, and P14: postoperative days 3, 7, and 14, respectively. C: control; S: surgery; S þ I: surgery þ CX3CL1. IL-1b, interleukin 1 beta; CX3CL1, C-X3-C motif chemokine ligand 1; SEM, standard error of the mean.

Article Snippet: A microsyringe drive (BASi, West Lafayette, IN) operated with a programmable controller (Bee Hive Controller; BASi) delivered recombinant rat CX3CL1 (5 mg in 0.9% NaCl, 5 mL, R&D Systems, Minneapolis, MN) to the animals at a rate of 1 mL/minute in the CX3CL1 group or 0.9% NaCl solution (5 mL) to the animals in the control group.

Techniques: Amplification, Expressing, Control

Journal: Journal of neuropathology and experimental neurology

Article Title: Surgical Trauma Exacerbates Cognitive Deficits and Neuroinflammation in Aged Rats: The Role of CX3CL1-CX3CR1 Signaling.

doi: 10.1093/jnen/nly051

Figure Lengend Snippet: FIGURE 9. BDNF protein expression was significantly downregulated following surgical challenge in the brains of the aged rats on postoperative days 3 and 7. The levels of BDNF had a reduction in the adult rats on postoperative day 3 and recovered on day 7. A lower level of BDNF was observed in the aged surgery group on postoperative day 3 compared with the day-matched adult surgery group. Treatment with CX3CL1 increased the level of BDNF compared with the day- matched aged surgery rats on postoperative day 3. The results are represented as the mean 6 SEM. *p < 0.05, **p < 0.001 versus the day-matched and age-matched control group; #p < 0.05 versus the day-matched and age-matched surgery group; &&p < 0.001 versus the day-matched adult surgery group. P3, P7, and P14: postoperative days 3, 7, and 14, respectively. C: control; S: surgery; S þ I: surgery þ CX3CL1. BDNF, brain derived neurotrophic factor; CX3CL1, C-X3-C motif chemokine ligand 1; SEM, standard error of the mean.

Article Snippet: A microsyringe drive (BASi, West Lafayette, IN) operated with a programmable controller (Bee Hive Controller; BASi) delivered recombinant rat CX3CL1 (5 mg in 0.9% NaCl, 5 mL, R&D Systems, Minneapolis, MN) to the animals at a rate of 1 mL/minute in the CX3CL1 group or 0.9% NaCl solution (5 mL) to the animals in the control group.

Techniques: Expressing, Control, Derivative Assay

Journal: Journal of neuropathology and experimental neurology

Article Title: Surgical Trauma Exacerbates Cognitive Deficits and Neuroinflammation in Aged Rats: The Role of CX3CL1-CX3CR1 Signaling.

doi: 10.1093/jnen/nly051

Figure Lengend Snippet: FIGURE 8. The mRNA levels of M2 phenotype marker Arg1 were downregulated in both adult and aged rats. Age amplified surgery-induced Arg1 downregulation on postoperative day 3 compared with the day-matched adult surgery group. The infusion of CX3CL1 significantly increased the level of Arg1 mRNA on postoperative day 3. The results are represented as the mean 6 SEM. **p < 0.001 versus the day-matched and age- matched control group; ##p < 0.001 versus the day-matched and age-matched surgery group; &p < 0.05 versus the day- matched adult surgery group. P3, P7, and P14: postoperative days 3, 7, and 14, respectively. Arg1, arginase-1; CX3CL1, C-X3-C motif chemokine ligand 1; SEM, standard error of the mean.

Article Snippet: A microsyringe drive (BASi, West Lafayette, IN) operated with a programmable controller (Bee Hive Controller; BASi) delivered recombinant rat CX3CL1 (5 mg in 0.9% NaCl, 5 mL, R&D Systems, Minneapolis, MN) to the animals at a rate of 1 mL/minute in the CX3CL1 group or 0.9% NaCl solution (5 mL) to the animals in the control group.

Techniques: Marker, Amplification, Control