TY - JOUR AU - de Oliveira Miranda, Camila AU - Hegedűs, Krisztina AU - Kis, Nikoletta Gréta AU - Antal, Miklós TI - Synaptic Targets of Glycinergic Neurons in Laminae I–III of the Spinal Dorsal Horn JF - INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES J2 - INT J MOL SCI VL - 24 PY - 2023 IS - 8 SN - 1661-6596 DO - 10.3390/ijms24086943 UR - https://m2.mtmt.hu/api/publication/33764688 ID - 33764688 AB - A great deal of evidence supports the inevitable importance of spinal glycinergic inhibition in the development of chronic pain conditions. However, it remains unclear how glycinergic neurons contribute to the formation of spinal neural circuits underlying pain-related information processing. Thus, we intended to explore the synaptic targets of spinal glycinergic neurons in the pain processing region (laminae I–III) of the spinal dorsal horn by combining transgenic technology with immunocytochemistry and in situ hybridization accompanied by light and electron microscopy. First, our results suggest that, in addition to neurons in laminae I–III, glycinergic neurons with cell bodies in lamina IV may contribute substantially to spinal pain processing. On the one hand, we show that glycine transporter 2 immunostained glycinergic axon terminals target almost all types of excitatory and inhibitory interneurons identified by their neuronal markers in laminae I–III. Thus, glycinergic postsynaptic inhibition, including glycinergic inhibition of inhibitory interneurons, must be a common functional mechanism of spinal pain processing. On the other hand, our results demonstrate that glycine transporter 2 containing axon terminals target only specific subsets of axon terminals in laminae I–III, including nonpeptidergic nociceptive C fibers binding IB4 and nonnociceptive myelinated A fibers immunoreactive for type 1 vesicular glutamate transporter, indicating that glycinergic presynaptic inhibition may be important for targeting functionally specific subpopulations of primary afferent inputs. LA - English DB - MTMT ER - TY - JOUR AU - Ducza, László AU - Gajtkó, Andrea AU - Hegedűs, Krisztina AU - Bakk, Erzsébet AU - Kis, Nikoletta Gréta AU - Gaál, Botond Ágoston AU - Takács, Roland Ádám AU - Szűcs, Péter AU - Matesz, Klára AU - Szentesiné Holló, Krisztina TI - Neuronal P2X4 receptor may contribute to peripheral inflammatory pain in rat spinal dorsal horn JF - FRONTIERS IN MOLECULAR NEUROSCIENCE J2 - FRONT MOL NEUROSCI VL - 16 PY - 2023 SN - 1662-5099 DO - 10.3389/fnmol.2023.1115685 UR - https://m2.mtmt.hu/api/publication/33682003 ID - 33682003 AB - Abstract Intense inflammation may result in pain, which manifests as spinal central sensitisation. There is growing evidence that purinergic signaling plays a pivotal role in the orchestration of pain processing. Over the last decade the ionotropic P2X purino receptor 4 (P2X4) got into spotlight in neuropathic disorders, however its precise spinal expression was scantily characterised during inflammatory pain. Thus, we intended to analyse the receptor distribution within spinal dorsal horn and lumbar dorsal root ganglia (DRG) of rats suffering in inflammatory pain induced by complete Freund adjuvant (CFA). Methods CFA- induced peripheral inflammation was validated by mechanical and thermal behavioural tests. In order to ensure about the putative alteration of spinal P2X4 receptor gene expression qPCR reactions were designed, followed by immunoperoxidase and Western blot experiments to assess changes at a protein level. Colocalisation of P2X4 with neuronal and glial markers was investigated by double immunofluorescent labelings, which were subsequently analysed with IMARIS software. Transmission electronmicroscopy was applied to study the ultrastructural localisation of the receptor. Concurrently, in lumbar DRG cells similar methodology has been carried out to complete our observations. Results The figures of mechanical and thermal behavioural tests proved the establishment of CFA- induced inflammatory pain. We observed significant enhancement of P2X4 transcript level within the spinal dorsal horn three days upon CFA administration. Elevation of P2X4 immunoreactivity within Rexed lamina I-II of the spinal gray matter was synchronous with mRNA expression, and confirmed by protein blotting. According to IMARIS analysis the robust protein increase was mainly detected on primary afferent axontermini and GFAP-labelled astrocyte membrane compartments within the spinal dorsal horn, but not on postsynaptic dendrites was also validated ultrastructurally. Furthermore, lumbar DRG analysis demonstrated that peptidergic and non-peptidergic nociceptive subsets of ganglia cells were also abundantly positive for P2X4 receptor in CFA model. Conclusions Here we provide novel evidence about involvement of neuronal and glial P2X4 receptor in the establishment of inflammatory pain. LA - English DB - MTMT ER - TY - JOUR AU - Balázs, Ervin AU - Hegedűs, Krisztina AU - Divéki, Zoltán TI - The unique carnation stunt-associated pararetroviroid JF - VIRUS RESEARCH J2 - VIRUS RES VL - 312 PY - 2022 SN - 0168-1702 DO - 10.1016/j.virusres.2022.198709 UR - https://m2.mtmt.hu/api/publication/32707412 ID - 32707412 N1 - Centre for Agricultural Research, Eötvö Loránd Research Network (ELKH), Brunszvik u. 2, Martonvásár, 2462, Hungary Department of Immunology, Eötvös Loránd University, Pázmány Péter sétány 1/C, Budapest, 1117, Hungary Cited By :1 Export Date: 9 February 2023 CODEN: VIRED Correspondence Address: Balázs, E.; Centre for Agricultural Research, Brunszvik u. 2, Hungary; email: balazs.ervin@atk.hu LA - English DB - MTMT ER - TY - JOUR AU - de Oliveira Miranda, Camila AU - Hegedűs, Krisztina AU - Wildner, Hendrik AU - Zeilhofer, Hanns Ulrich AU - Antal, Miklós TI - Morphological and neurochemical characterization of glycinergic neurons in laminae I-IV of the mouse spinal dorsal horn JF - JOURNAL OF COMPARATIVE NEUROLOGY J2 - J COMP NEUROL VL - 530 PY - 2022 IS - 3 SP - 607 EP - 626 PG - 20 SN - 0021-9967 DO - 10.1002/cne.25232 UR - https://m2.mtmt.hu/api/publication/32219371 ID - 32219371 AB - A growing body of experimental evidence shows that glycinergic inhibition plays vital roles in spinal pain processing. In spite of this, however, our knowledge about the morphology, neurochemical characteristics, and synaptic relations of glycinergic neurons in the spinal dorsal horn is very limited. The lack of this knowledge makes our understanding about the specific contribution of glycinergic neurons to spinal pain processing quite vague. Here we investigated the morphology and neurochemical characteristics of glycinergic neurons in laminae I-IV of the spinal dorsal horn using a GlyT2::CreERT2-tdTomato transgenic mouse line. Confirming previous reports, we show that glycinergic neurons are sparsely distributed in laminae I-II, but their densities are much higher in lamina III and especially in lamina IV. First in the literature, we provide experimental evidence indicating that in addition to neurons in which glycine colocalizes with GABA, there are glycinergic neurons in laminae I-II that do not express GABA and can thus be referred to as glycine-only neurons. According to the shape and size of cell bodies and dendritic morphology, we divided the tdTomato-labeled glycinergic neurons into three and six morphological groups in laminae I-II and laminae III-IV, respectively. We also demonstrate that most of the glycinergic neurons co-express neuronal nitric oxide synthase, parvalbumin, the receptor tyrosine kinase RET, and the retinoic acid-related orphan nuclear receptor beta (ROR beta), but there might be others that need further neurochemical characterization. The present findings may foster our understanding about the contribution of glycinergic inhibition to spinal pain processing. LA - English DB - MTMT ER - TY - JOUR AU - Ducza, László AU - Szűcs, Péter AU - Hegedűs, Krisztina AU - Bakk, Erzsébet AU - Gajtkó, Andrea AU - Wéber, Ildikó AU - Szentesiné Holló, Krisztina TI - NLRP2 Is Overexpressed in Spinal Astrocytes at the Peak of Mechanical Pain Sensitivity during Complete Freund Adjuvant-Induced Persistent Pain JF - INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES J2 - INT J MOL SCI VL - 22 PY - 2021 IS - 21 PG - 18 SN - 1661-6596 DO - 10.3390/ijms222111408 UR - https://m2.mtmt.hu/api/publication/32474149 ID - 32474149 N1 - 323780 LA - English DB - MTMT ER - TY - JOUR AU - Petri, László AU - Szijj, Péter A. AU - Kelemen, Ádám AU - Imre, Timea AU - Gömöry, Ágnes AU - Lee, Maximillian T. W. AU - Hegedűs, Krisztina AU - Ábrányi-Balogh, Péter AU - Chudasama, Vijay AU - Keserű, György Miklós TI - Cysteine specific bioconjugation with benzyl isothiocyanates JF - RSC ADVANCES J2 - RSC ADV VL - 10 PY - 2020 IS - 25 SP - 14928 EP - 14936 PG - 9 SN - 2046-2069 DO - 10.1039/d0ra02934c UR - https://m2.mtmt.hu/api/publication/31294506 ID - 31294506 N1 - Funding Agency and Grant Number: National Office of Research, Development and Innovation [2018-1.3.1-VKE-2018-00032, NKFIH PD124598]; Wellcome TrustWellcome TrustEuropean Commission; Hungarian Ministry for Innovation and Technology Funding text: This work has been supported by the National Office of Research, Development and Innovation (2018-1.3.1-VKE-2018-00032, NKFIH PD124598). We gratefully acknowledge the spectrophotometry measurements to Denes Sovari and the 15N-HSQC NMR measurements to Gyula Palfy and Andras Perczel. We gratefully acknowledge the Wellcome Trust for funding P. S. The flow cytometry facility is operated in ELTE Thematic Excellence Programme supported by the Hungarian Ministry for Innovation and Technology. LA - English DB - MTMT ER - TY - JOUR AU - Takáts, Szabolcs AU - Glatz, Gábor AU - Szenci, Győző AU - Boda, Attila AU - Horváth V., Gábor AU - Hegedűs, Krisztina AU - Kovács, Attila Lajos AU - Juhász, Gábor TI - Non-canonical role of the SNARE protein Ykt6 in autophagosome-lysosome fusion. JF - PLOS GENETICS J2 - PLOS GENET VL - 14 PY - 2018 IS - 4 PG - 23 SN - 1553-7390 DO - 10.1371/journal.pgen.1007359 UR - https://m2.mtmt.hu/api/publication/3364574 ID - 3364574 N1 - Funding Agency and Grant Number: Hungarian Academy of SciencesHungarian Academy of Sciences [LP-2014/2, PPD-003/2016]; National Research, Development and Innovation Office of Hungary [GINOP-2.32-15-2010-00006, GINOP-2.32-15-2010-00032, K119842, ERC-HU123410, KH125108] Funding text: This work was funded by the Hungarian Academy of Sciences (LP-2014/2 to GJ; PPD-003/2016 to ST) and National Research, Development and Innovation Office of Hungary (GINOP-2.32-15-2010-00006 and -00032, K119842 and ERC-HU123410 to GJ and KH125108 for ST). The funders had no role in study design, data collection and, analysis, decision to publish, or preparation of the manuscript. Funder websites are: http://mta.hu/http://nkfih.gov.hu/ AB - The autophagosomal SNARE Syntaxin17 (Syx17) forms a complex with Snap29 and Vamp7/8 to promote autophagosome-lysosome fusion via multiple interactions with the tethering complex HOPS. Here we demonstrate that, unexpectedly, one more SNARE (Ykt6) is also required for autophagosome clearance in Drosophila. We find that loss of Ykt6 leads to large-scale accumulation of autophagosomes that are unable to fuse with lysosomes to form autolysosomes. Of note, loss of Syx5, the partner of Ykt6 in ER-Golgi trafficking does not prevent autolysosome formation, pointing to a more direct role of Ykt6 in fusion. Indeed, Ykt6 localizes to lysosomes and autolysosomes, and forms a SNARE complex with Syx17 and Snap29. Interestingly, Ykt6 can be outcompeted from this SNARE complex by Vamp7, and we demonstrate that overexpression of Vamp7 rescues the fusion defect of ykt6 loss of function cells. Finally, a point mutant form with an RQ amino acid change in the zero ionic layer of Ykt6 protein that is thought to be important for fusion-competent SNARE complex assembly retains normal autophagic activity and restores full viability in mutant animals, unlike palmitoylation or farnesylation site mutant Ykt6 forms. As Ykt6 and Vamp7 are both required for autophagosome-lysosome fusion and are mutually exclusive subunits in a Syx17-Snap29 complex, these data suggest that Vamp7 is directly involved in membrane fusion and Ykt6 acts as a non-conventional, regulatory SNARE in this process. LA - English DB - MTMT ER - TY - JOUR AU - Csizmadia, Tamás AU - Lőrincz, Péter AU - Hegedűs, Krisztina AU - Szeplaki, S AU - Lőw, Péter AU - Juhász, Gábor TI - Molecular mechanisms of developmentally programmed crinophagy in Drosophila. JF - JOURNAL OF CELL BIOLOGY J2 - J CELL BIOL VL - 217 PY - 2018 IS - 1 SP - 361 EP - 374 PG - 14 SN - 0021-9525 DO - 10.1083/jcb.201702145 UR - https://m2.mtmt.hu/api/publication/3287984 ID - 3287984 N1 - Department of Anatomy, Cell and Developmental Biology, Eötvös Loránd University, Budapest, Hungary Institute of Genetics, Biological Research Centre, Hungarian Academy of Sciences, Szeged, Hungary Cited By :8 Export Date: 9 September 2019 CODEN: JCLBA Correspondence Address: Juhász, G.; Department of Anatomy, Cell and Developmental Biology, Eötvös Loránd UniversityHungary; email: szmrt@elte.hu Department of Anatomy, Cell and Developmental Biology, Eötvös Loránd University, Budapest, Hungary Institute of Genetics, Biological Research Centre, Hungarian Academy of Sciences, Szeged, Hungary Cited By :11 Export Date: 18 October 2019 CODEN: JCLBA Correspondence Address: Juhász, G.; Department of Anatomy, Cell and Developmental Biology, Eötvös Loránd UniversityHungary; email: szmrt@elte.hu Department of Anatomy, Cell and Developmental Biology, Eötvös Loránd University, Budapest, Hungary Institute of Genetics, Biological Research Centre, Hungarian Academy of Sciences, Szeged, Hungary Cited By :11 Export Date: 22 October 2019 CODEN: JCLBA Correspondence Address: Juhász, G.; Department of Anatomy, Cell and Developmental Biology, Eötvös Loránd UniversityHungary; email: szmrt@elte.hu AB - At the onset of metamorphosis, Drosophila salivary gland cells undergo a burst of glue granule secretion to attach the forming pupa to a solid surface. Here, we show that excess granules evading exocytosis are degraded via direct fusion with lysosomes, a secretory granule-specific autophagic process known as crinophagy. We find that the tethering complex HOPS (homotypic fusion and protein sorting); the small GTPases Rab2, Rab7, and its effector, PLEKHM1; and a SNAP receptor complex consisting of Syntaxin 13, Snap29, and Vamp7 are all required for the fusion of secretory granules with lysosomes. Proper glue degradation within lysosomes also requires the Uvrag-containing Vps34 lipid kinase complex and the v-ATPase proton pump, whereas Atg genes involved in macroautophagy are dispensable for crinophagy. Our work establishes the molecular mechanism of developmentally programmed crinophagy in Drosophila and paves the way for analyzing this process in metazoans. LA - English DB - MTMT ER - TY - CONF AU - Csizmadia, Tamás AU - Lőrincz, Péter AU - Hegedűs, Krisztina AU - Széplaki, Szilvia AU - Lőw, Péter AU - Juhász, Gábor ED - EDRC, null TI - Developmentally programmed secretory granule degradation in the Drosophila late larval and early pupal salivary gland cells T2 - EDRC 2017 C1 - Harrogate PY - 2017 PG - 1 UR - https://m2.mtmt.hu/api/publication/3363929 ID - 3363929 LA - English DB - MTMT ER - TY - JOUR AU - Nagy, Péter AU - Szatmári, Zsuzsanna AU - Sándor, Gyöngyvér Orsolya AU - Lippai, Mónika AU - Hegedűs, Krisztina AU - Juhász, Gábor TI - Drosophila Atg16 promotes enteroendocrine cell differentiation via regulation of intestinal Slit/Robo signaling. JF - DEVELOPMENT J2 - DEVELOPMENT VL - 144 PY - 2017 IS - 21 SP - 3990 EP - 4001 PG - 12 SN - 0950-1991 DO - 10.1242/dev.147033 UR - https://m2.mtmt.hu/api/publication/3278744 ID - 3278744 N1 - These authors contributed equally to this work: Péter Nagy, Zsuzsanna Szatmári AB - Genetic variations of Atg16L1, Slit and Rab19 predispose to the development of inflammatory bowel disease (IBD), but the relationship of these mutations is unclear. Here we show that in Drosophila guts lacking the WD40 domain of Atg16, pre-enteroendocrine cells (pre-EEs) accumulate that fail to differentiate into properly functioning secretory EEs. Mechanistically, loss of Atg16 or its binding partner Rab19 impairs Slit production, which normally inhibits EE generation by activating Robo signaling in stem cells. Importantly, loss of Atg16 or decreased Slit/Robo signaling trigger an intestinal inflammatory response. Surprisingly, analysis of Rab19 and domain-specific Atg16 mutants indicates that their stem cell niche regulatory function is independent of autophagy. Our study reveals how mutations in these different genes may contribute to IBD. LA - English DB - MTMT ER -