http://repo.lib.semmelweis.hu//handle/123456789/25 2026-10-14T22:25:18Z http://repo.lib.semmelweis.hu//handle/123456789/10455 Mapping chemoattractant gradients in live tissues Vámosi, Boldizsár; Enyedi, Balázs 2026-01-01T00:00:00Z http://repo.lib.semmelweis.hu//handle/123456789/10457 Hydrogen peroxide production by epidermal dual oxidase 1 regulates nociceptive sensory signals Pató, Anna; Bölcskei, Kata; Donkó, Ágnes; Kaszás, Diána; Boros, Melinda; Bodrogi, Lilla; Várady, György; Pape, Veronika; Roux, Benoit; Enyedi, Balázs; Helyes, Zzutzanna; Watt, F.M.; Sirokmány, Gábor; Geiszt, Miklós 2023-01-01T00:00:00Z http://repo.lib.semmelweis.hu//handle/123456789/10458 A genetically encoded sensor for visualizing leukotriene B4 gradients in vivo Tamás, Szimonetta Xénia; Roux, Benoit Thomas; Vámosi, Boldizsár; Dehne, Fabian Gregor; Török, Anna; Fazekas, László; Enyedi, Balázs Leukotriene B 4 (LTB 4 ) is a potent lipid chemoattractant driving inflammatory responses during host defense, allergy, autoimmune and metabolic diseases. Gradients of LTB 4 orchestrate leukocyte recruitment and swarming to sites of tissue damage and infection. How LTB 4 gradients form and spread in live tissues to regulate these processes remains largely elusive due to the lack of suitable tools for monitoring LTB 4 levels in vivo. Here, we develop GEM-LTB 4 , a genetically encoded green fluorescent LTB 4 biosensor based on the human G-protein-coupled receptor BLT1. GEM-LTB 4 shows high sensitivity, specificity and a robust fluorescence increase in response to LTB 4 without affecting downstream signaling pathways. We use GEM-LTB 4 to measure ex vivo LTB 4 production of murine neutrophils. Transgenic expression of GEM-LTB 4 in zebrafish allows the real-time visualization of both exogenously applied and endogenously produced LTB 4 gradients. GEM-LTB 4 thus serves as a broadly applicable tool for analyzing LTB 4 dynamics in various experimental systems and model organisms. 2023-01-01T00:00:00Z http://repo.lib.semmelweis.hu//handle/123456789/10459 Optimization of the Heterologous Expression of the Cannabinoid Type-1 (CB1) Receptor Horváth, Viktória Bea; Soltész-Katona, Eszter; Wisniewski, Éva; Rajki, Anikó; Halász, E.; Enyedi, Balázs; Hunyady, László; Tóth, András; Szanda, Gergő The G protein-coupled type 1 cannabinoid receptor (CB1R) mediates virtually all classic cannabinoid effects, and both its agonists and antagonists hold major therapeutic potential. Heterologous expression of receptors is vital for pharmacological research, however, overexpression of these proteins may fundamentally alter their localization pattern, change the signalling partner preference and may also spark artificial clustering. Additionally, recombinant CB1Rs are prone to intense proteasomal degradation, which may necessitate substantial modifications, such as N-terminal truncation or signal sequence insertion, for acceptable cell surface expression. We report here that tuning down the expression intensity of the full-length CB1R reduces proteasomal degradation and offers receptor levels that are comparable to those of endogenous CB1 receptors. As opposed to high-efficiency expression with conventional promoters, weak promoter-driven CB1R expression provides ERK 1/2 and p38 MAPK signalling that closely resemble the activity of endogenous CB1Rs. Moreover, weakly expressed CB1R variants exhibit plasma membrane localization, preserve canonical Gi-signalling but prevent CB1R-Gs coupling observed with high-expression variants. Based on these findings, we propose that lowering the expression level of G protein-coupled receptors should always be considered in heterologous expression systems in order to reduce the pressure on the proteasomal machinery and to avoid potential signalling artefacts. Copyright © 2021 Horváth, Soltész-Katona, Wisniewski, Rajki, Halász, Enyedi, Hunyady, Tóth and Szanda. 2021-01-01T00:00:00Z