Journal: Journal of Neurochemistry

Article Title: Complement C3a and C5a Receptors Are Presented in Mouse Sciatic and Human Sural Nerves and Selectively Modulate the Neuronal Function of Large‐Caliber Fibers in Mice

doi: 10.1111/jnc.70129

Figure Lengend Snippet: C3aR is localized to the paranodal region. Longitudinal and cross‐sections of human sural nerves were analyzed by immunofluorescence using an anti‐C3aR antibody (red, A, D). The paranodal region was stained with an anti‐Caspr antibody (green, B), and the axons were stained with an anti‐peripherin antibody (green, E). Double‐stained images are merged (C, F). Teased mouse sciatic fibers were stained for C3aR (red, G, J, M), Caspr (green, H, K), and peripherin (green, N). The white arrow points to the Schmidt–Lanterman incisures (G). Double‐stained images are merged (I, L, O). Caspr‐contactin‐associated protein; n = 10 mice; Scale bar, 20 μM.

Article Snippet: The C3aR agonist TLQP‐21 (500 nM, Cat. No. HY‐P1345A, MedChemExpress, NJ, USA), the C3aR antagonist JR14‐a (100 nM, Cat. No. HY‐138161, MedChemExpress) and the combination were applied at the same concentrations as described above.

Techniques: Immunofluorescence, Staining

Journal: Journal of Neurochemistry

Article Title: Complement C3a and C5a Receptors Are Presented in Mouse Sciatic and Human Sural Nerves and Selectively Modulate the Neuronal Function of Large‐Caliber Fibers in Mice

doi: 10.1111/jnc.70129

Figure Lengend Snippet: mRNA and protein expression in the mouse sciatic nerve. Quantitative PCR (qPCR) analysis and western blotting were performed on sciatic nerves from 13‐week‐old mice ( n = 6 nerves). Intrinsic expression of the complement receptors and factors is seen (A). The outcomes are presented relative to hypoxanthine guanine phosphoribosyltransferase expression using the 2 −ΔCT method calculation method. Proteins C3aR and C5aR1 are present in mouse sciatic nerves ( n = 6 nerves) (B, C). HPRT‐hypoxanthine guanine phosphoribosyltransferase; ** p < 0.01.

Article Snippet: The C3aR agonist TLQP‐21 (500 nM, Cat. No. HY‐P1345A, MedChemExpress, NJ, USA), the C3aR antagonist JR14‐a (100 nM, Cat. No. HY‐138161, MedChemExpress) and the combination were applied at the same concentrations as described above.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot

Journal: Journal of Neurochemistry

Article Title: Complement C3a and C5a Receptors Are Presented in Mouse Sciatic and Human Sural Nerves and Selectively Modulate the Neuronal Function of Large‐Caliber Fibers in Mice

doi: 10.1111/jnc.70129

Figure Lengend Snippet: TLQP‐21 C3aR agonist‐induced neuronal sensitivity. Representative recruitment curves from a control nerve (A) and a nerve treated with the C3aR agonist (B) are shown. TLQP‐21‐treated nerves responded to a lower stimulation current compared to control nerves, prevented by the application of the antagonist (C). EC50 is presented as the EC50 plus 95% CI (profile likelihood) (D). Control n = 10; TLQP‐21 n = 13; JR14a n = 7; TLQP‐21 JR14a n = 10. **** p < 0.0001.

Article Snippet: The C3aR agonist TLQP‐21 (500 nM, Cat. No. HY‐P1345A, MedChemExpress, NJ, USA), the C3aR antagonist JR14‐a (100 nM, Cat. No. HY‐138161, MedChemExpress) and the combination were applied at the same concentrations as described above.

Techniques: Control

Journal: Journal of Neurochemistry

Article Title: Complement C3a and C5a Receptors Are Presented in Mouse Sciatic and Human Sural Nerves and Selectively Modulate the Neuronal Function of Large‐Caliber Fibers in Mice

doi: 10.1111/jnc.70129

Figure Lengend Snippet: The C3aR agonist modulates excitatory neuronal activity. Panel (A) shows a representative example of the refractory response in a control nerve (black, top) compared with a nerve treated with the C3aR agonist (green, bottom). The application of the C3aR agonist led to a reduction in the refractory period, which was prevented by the application of an antagonist (B). In Panel (C), representative examples of the response to high‐frequency stimulation (600–1000 Hz) are presented, comparing a control nerve (black, top) with a nerve following C3aR agonist application (green, bottom). In panel (D), representative examples of the slope at 1000 Hz are presented for a control nerve (black, left) compared with a C3aR agonist‐treated nerve (green, right). The slope in the C3aR agonist‐treated nerves was less steep than that in the control nerves, with the antagonist preventing this excitatory response (E). In panel (F), representative examples of the compound action potential response to 1000 Hz are shown (control: black, top; C3aR agonist: green, bottom). The difference between consecutive compound action potentials was greater in the control nerves than in the C3aR agonist‐treated nerves (G). Control n = 9, TLQP‐21 n = 13, and TLQP‐21 + JR14a n = 10, **** p < 0.0001.

Article Snippet: The C3aR agonist TLQP‐21 (500 nM, Cat. No. HY‐P1345A, MedChemExpress, NJ, USA), the C3aR antagonist JR14‐a (100 nM, Cat. No. HY‐138161, MedChemExpress) and the combination were applied at the same concentrations as described above.

Techniques: Activity Assay, Control

Journal: Journal of Neurochemistry

Article Title: Complement C3a and C5a Receptors Are Presented in Mouse Sciatic and Human Sural Nerves and Selectively Modulate the Neuronal Function of Large‐Caliber Fibers in Mice

doi: 10.1111/jnc.70129

Figure Lengend Snippet: C3aR neuronal hyperexcitability through potassium channel mediation. The slope of the response to high frequencies was similar for low KCl and low KCl + TLQP‐21 (A) Control = white; TLQP‐21 = green. A decreased response to high‐frequency firing rates was observed in the TEA and TEA‐TLQP‐21 nerves (B). Control n = 9; TLQP‐21 n = 12; Low KCl n = 4; Low KCl + TLQP‐21 n = 5; TEA n = 4; TEA + TLQP‐21 n = 6; TEA‐ tetraethyl ammonium chloride; ** p < 0.01.

Article Snippet: The C3aR agonist TLQP‐21 (500 nM, Cat. No. HY‐P1345A, MedChemExpress, NJ, USA), the C3aR antagonist JR14‐a (100 nM, Cat. No. HY‐138161, MedChemExpress) and the combination were applied at the same concentrations as described above.

Techniques: Control