A mozgató működésekért felelős gerincvelői neuronhálózatok morfológiai, fiziológiai vizsgálata és számítógépes szimulációja = Morphological, physiological investigations and computer simulation of spinal neuronal network involved in locomotion.

1. Fiziológiai kísérletekben jelentős különbségeket találtak propriospinalis axon - motoneuron (MN) kapcsolatokban a MN-ból elvezetett EPSP amplitúdójában a béka gerincvelőben. Modellkísérletekkel bizonyítottuk, hogy a kapcsolatok között mért eltéréseket több tényező magyarázhatja: a szinapszisok elhelyezkedése, a dendrit kitüremkedések, nem-lineáris PSP szummáció illetve a PSP-ok szómára érkezéséhez szükséges késési idők eltérései. 2. A Xenopus embrió úszómozgásáért felelős központi ritmusgeneráló hálózatában azt vizsgáltuk, hogyan vezethet egy rövid inger hosszú ideig tartó úszó mozgáshoz. Az utóagy-gerincvelő határának populációs modelljében azt találtuk, hogy az utóagyi neuronok közötti reciprok serkentő kapcsolatok felelősek lehetnek az említett jelenségért. Elképzelésünket a kísérletes eredmények megerősítik. 3. Patkány gerincvelőben vizsgáltuk a cink kolokalizációját gátló aminosavakkal. Eredményeink azt mutatták, hogy a gerincvelőben a cink tartalmú terminálisok elsősorban gátló karakterűek, amelyekben a GABA és glicin is megtalálható. 4. Újszülött patkány gerincvelőben elemeztük a commissuralis interneuronok (CIN) neurokémiai sajátságait és szinaptikus kapcsolatait. A jelölt CIN terminálisok több mint fele tartalmazott gátló aminosavat, míg a glutamátot tartalmazó terminálisok aránya 27% volt. A jelölt CIN terminálisok közvetlen kapcsolatot képeztek az ellenoldali motoneuronokkal és CIN-okkal. | 1. We investigated the postsynaptic factors that may contribute the high variability of synaptic efficacy in monosynaptically connected propriospinal axon-motoneuron pairs in the spinal cord of frog. We proved that differences in location of these synapses, dendritic protrusions, non-linear summation of PSPs, sizes of time windows for effective temporal summation and differences in delays of arrivals of PSPs to soma are all factors that may differentiate between high and low efficacy single fiber propriospinal connections found experimentally. 2. The central pattern generator for swimming in the Xenopus tadpole was investigated to find out how a brief stimulus can lead to prolonged swimming. By using large-scale population model of the hindbrain-spinal cord junction we found that reciprocal excitatory connections among hindbrain neurons may be responsible for this phenomenon. Our proposal has strong experimental support. 3. Colocalization of zinc with inhibitory neurotrasmitters was investigated in the rat spinal cord. 70% of zinc-containing terminals showed immunoreactivity for GABA and glycine. 4. In neonatal rats we investigated the axonal projection and neurotransmitter properties of spinal commissural interneurons (CIN). About half of the labelled CIN terminals contained GABA or glycine and one third proved to be excitatory. Many of CIN terminals made close appositions with motor neurons and also with CINs on the opposite side of the spinal cord

Nerve stretch injury induced pain pattern and changes in sensory ganglia in a clinically relevant model of limb-lengthening in rabbits.

We used a model of tibial lengthening in rabbits to study the postoperative pain pattern during limb-lengthening and morphological changes in the dorsal root ganglia (DRG), including alteration of substance P (SP) expression. Four groups of animals (naive; OG: osteotomised only group; SDG/FDG: slow/fast distraction groups, with 1 mm/3 mm lengthening a day, respectively) were used. Signs of increasing postoperative pain were detected until the 10(th) postoperative day in OG/SDG/FDG, then they decreased in OG but remained higher in SDG/FDG until the distraction finished, suggesting that the pain response is based mainly on surgical trauma until the 10(th) day, while the lengthening extended its duration and increased its intensity. The only morphological change observed in the DRGs was the presence of large vacuoles in some large neurons of OG/SDG/FDG. Cell size analysis of the S1 DRGs showed no cell loss in any of the three groups; a significant increase in the number of SP-positive large DRG cells in the OG; and a significant decrease in the number of SP-immunoreactive small DRG neurons in the SDG/FDG. Faster and larger distraction resulted in more severe signs of pain sensation, and further reduced the number of SP-positive small cells, compared to slow distraction

Pathology and glia type specific changes of the DPP4 activity in the spinal cord contributes to the development and maintenance of hyperalgesia and shapes opioid signalling in chronic pain states

Altered pain sensations such as hyperalgesia and allodynia are characteristic features of various chronic pain states, and remain difficult to treat. We have shown previously that spinal application of dipeptidyl peptidase 4 (DPP4) enzyme inhibitors results in a strong antihyperalgesic effect during inflammatory pain states. In this study we observed a low level of mRNA for DPP4 in the rat spinal dorsal horn in physiological conditions, which did not change significantly either in carrageenan-induced inflammatory or partial nerve ligation-generated neuropathic states. Although DPP4 protein was detected in neurons, astrocytes and microglia in naïve animals its expression significantly increased in astrocytes during inflammation and in microglia in neuropathic conditions. Intrathecal application of two DPP4 inhibitors the tripeptide isoleucin-prolin-isoleucin (IPI) and the antidiabetic drug vildagliptin resulted in robust opioid-dependent antihyperalgesic effect during inflammation and an opioid-independent effect in the Seltzer model. The opioid-mediated antihyperalgesic effect of IPI was exclusively related to mu-opioid receptors, while vildagliptin affected mainly delta-receptor activity, although mu- and kappa-receptors were also involved. Our results suggest a pathology and glia type specific changes of the DPP4 activity in the spinal cord which contributes to the development and maintenance of hyperalgesia and shapes opioid signalling

Similarity and dissimilarity in antinociceptive effects of dipeptidyl-peptidase 4 inhibitors, Diprotin A and vildagliptin in rat inflammatory pain models following spinal administration.

Dipeptidyl-peptidase 4 (DPP4) enzyme is involved in the degradation of many biologically active peptides including opioids. Its role in pain transmission is poorly elucidated. Recently we reported on the spinal antihyperalgesic effects of DPP4 inhibitors, Ile-Pro-Ile (Diprotin A) and vildagliptin in carrageenan-evoked acute inflammatory pain in rats. The present study investigated the effects of intrathecal (it.) diprotin A and vildagliptin in Complete Freund's Adjuvant- (CFA) and formalin induced pain in rats. The former assay can model the subchronic inflammatory pain condition and the later one reflects both acute tonic and inflammatory pain conditions. The involvement of opioid receptor (OR) subtypes, Y1-, and GLP1 receptors were also investigated. In CFA pain model it. diprotin A or vildagliptin dose-dependently inhibits hyperalgesia in ipsilateral while has no effect in contralateral paws. The peak effect was achieved 30 min following drug administration which was used for further analysis. Both compounds showed naltrexone reversible antihyperalgesia. Co-administration of OR-subtype-selective antagonists with diprotin A and vildagliptin revealed involvement of μ and δ > μ opioid receptors, respectively. Co-administered Y1 but not GLP1 receptor antagonists reversed the antihyperalgesic action of both DPP4 inhibitors. In touch-hypersensitivity both compounds were ineffective. In formalin test only diprotin A showed μ and δ OR-mediated antinociception and only in the 2nd phase. This effect was Y1 or GLP-1 receptor antagonist insensitive. In conclusion, diprotin A and vildagliptin display antinociception of different mechanisms of action in subchronic inflammatory pain. Furthermore, the spinal pain relay points of inflammatory pain of acute or subchronic conditions were more effectively affected by diprotin A than vildagliptin which needs future elucidation

Unique, Specific CART Receptor-Independent Regulatory Mechanism of CART(55-102) Peptide in Spinal Nociceptive Transmission and Its Relation to Dipeptidyl-Peptidase 4 (DDP4)

Cocaine- and amphetamine-regulated transcript (CART) peptides are involved in several physiological and pathological processes, but their mechanism of action is unrevealed due to the lack of identified receptor(s). We provided evidence for the antihyperalgesic effect of CART(55-102) by inhibiting dipeptidyl-peptidase 4 (DPP4) in astrocytes and consequently reducing neuroinflammation in the rat spinal dorsal horn in a carrageenan-evoked inflammation model. Both naturally occurring CART(55-102) and CART(62-102) peptides are present in the spinal cord. CART(55-102) is not involved in acute nociception but regulates spinal pain transmission during peripheral inflammation. While the full-length peptide with a globular motif contributes to hyperalgesia, its N-terminal inhibits this process. Although the anti-hyperalgesic effects of CART(55-102), CART(55-76), and CART(62-76) are blocked by opioid receptor antagonists in our inflammatory models, but not in neuropathic Seltzer model, none of them bind to any opioid or G-protein coupled receptors. DPP4 interacts with Toll-like receptor 4 (TLR4) signalling in spinal astrocytes and enhances the TLR4-induced expression of interleukin-6 and tumour necrosis factor alpha contributing to inflammatory pain. Depending on the state of inflammation, CART(55-102) is processed in the spinal cord, resulting in the generation of biologically active isoleucine-proline-isoleucine (IPI) tripeptide, which inhibits DPP4, leading to significantly decreased glia-derived cytokine production and hyperalgesia