TY - JOUR AU - Kovács, Kinga Dóra AU - Visnovitz, Tamás AU - Gerecsei, Tamás AU - Péter, Beatrix AU - Kurunczi, Sándor AU - Koncz, Anna AU - Németh, Krisztina AU - Lenzinger, Dorina AU - Visnovitzné Dr Vukman, Krisztina AU - Balogh, Anna AU - Rajmon, Imola AU - Lőrincz, Péter AU - Székács, Inna AU - Buzás, Edit Irén AU - Horváth, Róbert TI - Nanoinjection of extracellular vesicles to single live cells by robotic fluidic force microscopy JF - JOURNAL OF EXTRACELLULAR VESICLES J2 - J EXTRACELLULAR VESICL VL - 12 PY - 2023 IS - 12 PG - 16 SN - 2001-3078 DO - 10.1002/jev2.12388 UR - https://m2.mtmt.hu/api/publication/34425078 ID - 34425078 AB - In the past decade, extracellular vesicles (EVs) have attracted substantial interest in biomedicine. With progress in the field, we have an increasing understanding of cellular responses to EVs. In this Technical Report, we describe the direct nanoinjection of EVs into the cytoplasm of single cells of different cell lines. By using robotic fluidic force microscopy (robotic FluidFM), nanoinjection of GFP positive EVs and EV‐like particles into single live HeLa, H9c2, MDA‐MB‐231 and LCLC‐103H cells proved to be feasible. This injection platform offered the advantage of high cell selectivity and efficiency. The nanoinjected EVs were initially localized in concentrated spot‐like regions within the cytoplasm. Later, they were transported towards the periphery of the cells. Based on our proof‐of‐principle data, robotic FluidFM is suitable for targeting single living cells by EVs and may lead to information about intracellular EV cargo delivery at a single‐cell level. LA - English DB - MTMT ER - TY - JOUR AU - Vedelek, Viktor AU - Vedelek, Balázs AU - Lőrincz, Péter AU - Juhász, Gábor AU - Sinka, Rita TI - A comparative analysis of fruit fly and human glutamate dehydrogenases in Drosophila melanogaster sperm development JF - FRONTIERS IN CELL AND DEVELOPMENTAL BIOLOGY J2 - FRONT CELL DEV BIOL VL - 11 PY - 2023 PG - 16 SN - 2296-634X DO - 10.3389/fcell.2023.1281487 UR - https://m2.mtmt.hu/api/publication/34239207 ID - 34239207 N1 - Funding Agency and Grant Number: The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This work was supported by NKFIH (PD137914 to VV, K132155 to RS, and FK138851 to PL), National Research, Development, and Innovation Offi [PD137914, K132155, FK138851]; NKFIH [2022-2.1.1-NL-2022-00008, EKA 2022/045-P101-2, LP 2022-13/2022]; National Research, Development, and Innovation Office of Hungary (Biotechnology National Laboratory) Funding text: The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This work was supported by NKFIH (PD137914 to VV, K132155 to RS, and FK138851 to PL), National Research, Development, and Innovation Office of Hungary (Biotechnology National Laboratory 2022-2.1.1-NL-2022-00008 to GJ), and Eotvos Lorand University Excellence Fund (EKA 2022/045-P101-2), MTA-t (LP 2022-13/2022) to PL.r The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This work was supported by NKFIH (PD137914 to VV, K132155 to RS, and FK138851 to PL), National Research, Development, and Innovation Office of Hungary (Biotechnology National Laboratory 2022-2.1.1-NL-2022-00008 to GJ), and Eotvos Lorand University Excellence Fund (EKA 2022/045-P101-2), MTA-t (LP 2022-13/2022) to PL. AB - Glutamate dehydrogenases are enzymes that take part in both amino acid and energy metabolism. Their role is clear in many biological processes, from neuronal function to cancer development. The putative testis-specific Drosophila glutamate dehydrogenase, Bb8, is required for male fertility and the development of mitochondrial derivatives in spermatids. Testis-specific genes are less conserved and could gain new functions, thus raising a question whether Bb8 has retained its original enzymatic activity. We show that while Bb8 displays glutamate dehydrogenase activity, there are significant functional differences between the housekeeping Gdh and the testis-specific Bb8. Both human GLUD1 and GLUD2 can rescue the bb8 ms mutant phenotype, with superior performance by GLUD2. We also tested the role of three conserved amino acids observed in both Bb8 and GLUD2 in Gdh mutants, which showed their importance in the glutamate dehydrogenase function. The findings of our study indicate that Drosophila Bb8 and human GLUD2 could be novel examples of convergent molecular evolution. Furthermore, we investigated the importance of glutamate levels in mitochondrial homeostasis during spermatogenesis by ectopic expression of the mitochondrial glutamate transporter Aralar1, which caused mitochondrial abnormalities in fly spermatids. The data presented in our study offer evidence supporting the significant involvement of glutamate metabolism in sperm development. LA - English DB - MTMT ER - TY - JOUR AU - Maruzs, Tamás AU - Feil-Börcsök, Dalma AU - Lakatos, Enikő AU - Juhász, Gábor AU - Blastyák, András AU - Hargitai, Dávid AU - Jean, Steve AU - Lőrincz, Péter AU - Juhász, Gábor TI - Interaction of the sorting nexin 25 homologue Snazarus with Rab11 balances endocytic and secretory transport and maintains the ultrafiltration diaphragm in nephrocytes JF - MOLECULAR BIOLOGY OF THE CELL J2 - MOL BIOL CELL VL - 34 PY - 2023 IS - 9 PG - 14 SN - 1059-1524 DO - 10.1091/mbc.E22-09-0421 UR - https://m2.mtmt.hu/api/publication/34021340 ID - 34021340 N1 - Funding Agency and Grant Number: National Research Development and Innovation Office (NKFIH) of Hungary [PD135611, UNKP-22-2-III-ELTE-702, FK138851, Elvonal KKP129797]; National Laboratory of Biotechnology [2022-2.1.1-NL-2022-00008]; Eotvos Lorand University Excellence Fund [EKA 2022/045-P101-2]; National Academy of Scientist Education under the sponsorship of the Hungarian Ministry of Innovation and Technology [FEIF/646-4/2021-ITM_SZERZ] Funding text: We thank Szilvia Bozso and Monika Truszka for technical assistance. We thank Mike W. Henne for suggestions and for providing reagents and Michael Krahn and Thomas L. Schwarz for providing reagents. This work was supported by the National Research Development and Innovation Office (NKFIH) of Hungary with PD135611 grant to T.M., UNKP-22-2-III-ELTE-702 grant to D.H., FK138851 grant to P.L., and Elvonal KKP129797 and the National Laboratory of Biotechnology 2022-2.1.1-NL-2022-00008 grants to G.J. The work was also supported by the Eotvos Lorand University Excellence Fund (EKA 2022/045-P101-2) to P.L.. This research work was conducted with additional support from the National Academy of Scientist Education under the sponsorship of the Hungarian Ministry of Innovation and Technology (FEIF/646-4/2021-ITM_SZERZ). AB - Proper balance of exocytosis and endocytosis is important for the maintenance of plasma membrane lipid and protein homeostasis. This is especially critical in human podocytes and the podocyte-like Drosophila nephrocytes that both use a delicate diaphragm system with evolutionarily conserved components for ultrafiltration. Here we show that the sorting nexin 25 homolog Snazarus (Snz) binds to Rab11 and localizes to Rab11-positive recycling endosomes in Drosophila nephrocytes, unlike in fat cells where it is present in plasma membrane/lipid droplet/ER contact sites. Loss of Snz leads to redistribution of Rab11 vesicles from the cell periphery and increases endocytic activity in nephrocytes. These changes are accompanied by defects in diaphragm protein distribution that resemble those seen in Rab11 gain-of-function cells. Of note, co-overexpression of Snz rescues diaphragm defects in Rab11 overexpressing cells, whereas snz knockdown in Rab11 overexpressing nephrocytes or simultaneous knockdown of snz and tbc1d8b encoding a Rab11 GAP lead to massive expansion of the lacunar system that contains mislocalized diaphragm components: Sns and Pyd/ZO-1. We find that loss of Snz enhances while its overexpression impairs secretion, which, together with genetic epistasis analyses, suggest that Snz counteracts Rab11 to maintain the diaphragm via setting the proper balance of exocytosis and endocytosis. LA - English DB - MTMT ER - TY - JOUR AU - Koncz, Anna AU - Turiák, Lilla AU - Németh, Krisztina AU - Lenzinger, Dorina AU - Bárkai, Tünde AU - Lőrincz, Péter AU - Zelenyánszki, Helga AU - Visnovitzné Dr Vukman, Krisztina AU - Buzás, Edit Irén AU - Visnovitz, Tamás TI - Endoplasmin Is a Hypoxia-Inducible Endoplasmic Reticulum-Derived Cargo of Extracellular Vesicles Released by Cardiac Cell Lines JF - MEMBRANES (BASEL) J2 - MEMBRANES-BASEL VL - 13 PY - 2023 IS - 4 PG - 16 SN - 2077-0375 DO - 10.3390/membranes13040431 UR - https://m2.mtmt.hu/api/publication/33750753 ID - 33750753 N1 - Department of Genetics, Cell and Immunobiology, Semmelweis University, Üllői út 26, Budapest, 1085, Hungary Research Centre for Natural Sciences, Institute of Organic Chemistry, Magyar Tudósok Körútja 2, Budapest, 1117, Hungary ELKH-SE Translational Extracellular Vesicle Research Group, Nagyvárad tér 4, Budapest, 1085, Hungary Department of Anatomy, Cell and Developmental Biology, ELTE Eötvös Loránd University, Pázmány Péter sétány 1/c, Budapest, 1117, Hungary Department of Plant Physiology and Molecular Plant Biology, ELTE Eötvös Loránd University, Pázmány Péter sétány 1/c, Budapest, 1117, Hungary HCEMM-SU Extracellular Vesicle Research Group, Nagyvárad tér 4, Budapest, 1085, Hungary Export Date: 30 May 2023 Correspondence Address: Visnovitz, T.; Department of Genetics, Üllői út 26, Hungary; email: visnovitz.tamas@med.semmelweis-univ.hu AB - Cardiomyopathies are leading causes of human mortality. Recent data indicate that the cardiomyocyte-derived extracellular vesicles (EVs) released upon cardiac injury are present in circulation. This paper aimed to analyze EVs released under normal and hypoxic conditions by H9c2 (rat), AC16 (human) and HL1 (mouse) cardiac cell lines. Small (sEVs), medium (mEVs) and large EVs (lEVs) were separated from a conditioned medium by a combination of gravity filtration, differential centrifugation and tangential flow filtration. The EVs were characterized by microBCA, SPV lipid assay, nanoparticle tracking analysis, transmission and immunogold electron microscopy, flow cytometry and Western blotting. Proteomic profiles of the EVs were determined. Surprisingly, an endoplasmic reticulum chaperone, endoplasmin (ENPL, grp94 or gp96), was identified in the EV samples, and its association with EVs was validated. The secretion and uptake of ENPL was followed by confocal microscopy using GFP-ENPL fusion protein expressing HL1 cells. We identified ENPL as an internal cargo of cardiomyocyte-derived mEVs and sEVs. Based on our proteomic analysis, its presence in EVs was linked to hypoxia in HL1 and H9c2 cells, and we hypothesize that EV-associated ENPL may have a cardioprotective role by reducing cardiomyocyte ER stress. LA - English DB - MTMT ER - TY - JOUR AU - Rzepiel, Andrea AU - Horváth, Anna AU - Kutszegi, Nóra AU - Gézsi, András AU - C. Sági, Judit AU - Almási, Laura AU - Egyed, Bálint AU - Lőrincz, Péter AU - Visnovitz, Tamás AU - Kovács, Gábor AU - Szalai, Csaba AU - Semsei F, Ágnes AU - Erdélyi, Dániel TI - MiR-128-3p as blood based liquid biopsy biomarker in childhood acute lymphoblastic leukemia JF - MOLECULAR AND CELLULAR PROBES J2 - MOL CELL PROBE VL - 67 PY - 2023 PG - 7 SN - 0890-8508 DO - 10.1016/j.mcp.2023.101893 UR - https://m2.mtmt.hu/api/publication/33577132 ID - 33577132 N1 - Dept. of Paediatrics, Semmelweis University, Budapest, Hungary Dept. of Genetics, Cell- and Immunobiology, Semmelweis University, Budapest, Hungary Dept. of Measurement and Information Systems, Budapest University of Technology and Economics, Budapest, Hungary Institute of Genomic Medicine and Rare Disorders, Semmelweis University, Budapest, Hungary HCEMM-SE Molecular Oncohematology Research Group, 1st Dept. of Pathology and Experimental Cancer Research, Semmelweis University, Budapest, Hungary Dept. of Anatomy, Cell and Developmental Biology, ELTE Eötvös Loránd University, Budapest, Hungary Dept. of Plant Physiology and Molecular Plant Biology, ELTE Eötvös Loránd University, Budapest, Hungary Heim Pál Children Hospital, Budapest, Hungary Cited By :1 Export Date: 24 April 2023 CODEN: MCPRE Correspondence Address: Semsei, Á.F.; Dept. of Genetics, 1085 Üllői út 26, Hungary; email: semsei.agnes@med.semmelweis-univ.hu LA - English DB - MTMT ER - TY - JOUR AU - Boda, Attila AU - Varga, Luca Petra AU - Nagy, Anikó Zsuzsanna AU - Szenci, Győző AU - Csizmadia, Tamás AU - Lőrincz, Péter AU - Juhász, Gábor TI - Rab26 controls secretory granule maturation and breakdown in Drosophila JF - CELLULAR AND MOLECULAR LIFE SCIENCES J2 - CELL MOL LIFE SCI VL - 80 PY - 2023 IS - 1 PG - 15 SN - 1420-682X DO - 10.1007/s00018-022-04674-8 UR - https://m2.mtmt.hu/api/publication/33542161 ID - 33542161 N1 - Export Date: 17 January 2023 CODEN: CMLSF AB - At the onset of Drosophila metamorphosis, plenty of secretory glue granules are released from salivary gland cells and the glue is deposited on the ventral side of the forming (pre)pupa to attach it to a dry surface. Prior to this, a poorly understood maturation process takes place during which secretory granules gradually grow via homotypic fusions, and their contents are reorganized. Here we show that the small GTPase Rab26 localizes to immature (smaller, non-acidic) glue granules and its presence prevents vesicle acidification. Rab26 mutation accelerates the maturation, acidification and release of these secretory vesicles as well as the lysosomal breakdown (crinophagy) of residual, non-released glue granules. Strikingly, loss of Mon1, an activator of the late endosomal and lysosomal fusion factor Rab7, results in Rab26 remaining associated even with the large glue granules and a concomitant defect in glue release, similar to the effects of Rab26 overexpression. Our data thus identify Rab26 as a key regulator of secretory vesicle maturation that promotes early steps (vesicle growth) and inhibits later steps (lysosomal transport, acidification, content reorganization, release, and breakdown), which is counteracted by Mon1. LA - English DB - MTMT ER - TY - GEN AU - Gerecsei, Tamás AU - Visnovitz, Tamás AU - Kovács, Kinga Dóra AU - Péter, Beatrix AU - Kurunczi, Sándor AU - Koncz, Anna AU - Németh, Krisztina AU - Lenzinger, Dorina AU - Visnovitzné Dr Vukman, Krisztina AU - Lőrincz, Péter AU - Székács, Inna AU - Buzás, Edit Irén AU - Horváth, Róbert TI - Nanoinjection of fluorescent nanoparticles to single live cells by robotic fluidic force microscopy PY - 2022 UR - https://m2.mtmt.hu/api/publication/33589125 ID - 33589125 LA - English DB - MTMT ER - TY - JOUR AU - Nagy, Anikó Zsuzsanna AU - Szenci, Győző AU - Boda, Attila AU - Al-Lami, Muna Adnan Idan AU - Csizmadia, Tamás AU - Lőrincz, Péter AU - Juhász, Gábor AU - Lőw, Péter TI - Ecdysone receptor isoform specific regulation of secretory granule acidification in the larval Drosophila salivary gland JF - EUROPEAN JOURNAL OF CELL BIOLOGY J2 - EUR J CELL BIOL VL - 101 PY - 2022 IS - 4 SN - 0171-9335 DO - 10.1016/j.ejcb.2022.151279 UR - https://m2.mtmt.hu/api/publication/33190551 ID - 33190551 N1 - Department of Anatomy, Cell and Developmental Biology, Eötvös Loránd University, Budapest, Hungary Institute of Genetics, Biological Research Centre, Szeged, Hungary Cited By :1 Export Date: 19 September 2023 CODEN: EJCBD Correspondence Address: Juhász, G.; Biological Research Centre, Hungary; email: szmrt@elte.hu LA - English DB - MTMT ER - TY - JOUR AU - Hargitai, Dávid AU - Kenéz, Lili Anna AU - Al-Lami, Muna Adnan Idan AU - Szenci, Győző AU - Lőrincz, Péter AU - Juhász, Gábor TI - Autophagy controls Wolbachia infection upon bacterial damage and in aging Drosophila JF - FRONTIERS IN CELL AND DEVELOPMENTAL BIOLOGY J2 - FRONT CELL DEV BIOL VL - 10 PY - 2022 PG - 11 SN - 2296-634X DO - 10.3389/fcell.2022.976882 UR - https://m2.mtmt.hu/api/publication/33162692 ID - 33162692 N1 - Funding Agency and Grant Number: New National Excellence Program of the Ministry for Innovation and Technology; National Research, Development and Innovation Office (Hungary) [UNKP-21-2-II-ELTE- 703, UNKP-215-ELTE-1122]; Hungarian Academy of Sciences [PPD-222/2018, BO- 00514- 21-8]; National Research, Development and Innovation Office [FK 138851, GINOP-2.3.2-15- 2016-00006, GINOP-2.3.2-15- 2016-00032, K119842, KKP129797]; Eotvos Lorand University Excellence Fund [EKA 2022/ 045-P101-2] Funding text: This work was funded by the New National Excellence Program of the Ministry for Innovation and Technology and the National Research, Development and Innovation Office (Hungary) ( UNKP-21-2-II-ELTE- 703 to DH and UNKP-215-ELTE-1122 to PL), the Hungarian Academy of Sciences (PPD-222/2018 and BO- 00514- 21-8 to PL) and the National Research, Development and Innovation Office (FK 138851 to PL, GINOP-2.3.2-15- 2016-00006 and -00032, K119842, KKP129797 and National Biotechnology Laboratory to GJ) and Eotvos Lorand University Excellence Fund ( EKA 2022/ 045-P101-2 to PL). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. AB - Autophagy is a conserved catabolic process in eukaryotic cells that degrades intracellular components in lysosomes, often in an organelle-specific selective manner (mitophagy, ERphagy, etc). Cells also use autophagy as a defense mechanism, eliminating intracellular pathogens via selective degradation known as xenophagy. Wolbachia pipientis is a Gram-negative intracellular bacterium, which is one of the most common parasites on Earth affecting approximately half of terrestrial arthropods. Interestingly, infection grants the host resistance against other pathogens and modulates lifespan, so this bacterium resembles an endosymbiont. Here we demonstrate that Drosophila somatic cells normally degrade a subset of these bacterial cells, and autophagy is required for selective elimination of Wolbachia upon antibiotic damage. In line with these, Wolbachia overpopulates in autophagy-compromised animals during aging while its presence fails to affect host lifespan unlike in case of control flies. The autophagic degradation of Wolbachia thus represents a novel antibacterial mechanism that controls the propagation of this unique bacterium, behaving both as parasite and endosymbiont at the same time. LA - English DB - MTMT ER - TY - CONF AU - Lőrincz, Péter TI - Korai endoszomális SNARE komplexek vizsgálata és azonosítása T2 - Intézményi ÚNKP Konferencia 2022 PB - Eötvös Loránd Tudományegyetem (ELTE) C1 - Budapest PY - 2022 SP - 298 UR - https://m2.mtmt.hu/api/publication/33113442 ID - 33113442 LA - Hungarian DB - MTMT ER -