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Modulation of synaptic transmission in the development of painful states
Slepička, Jakub ; Paleček, Jiří (advisor) ; Hejnová, Lucie (referee)
My thesis introduces the topic of nociceptive signalisation and processes involved in the formation and spreading of neuropathic pain. This study focuses on the mechanisms of nociceptive synaptic transmission mechanisms in the level of spinal dorsal horn and its modulation by paclitaxel, a chemotherapeutic drug inducing neuropathic changes. The attention is put especially on the possibility of glial activity participation in paclitaxel side effects. This idea stems from the existing hypothesis of the functional connection between TLR4 and TRPV1 receptor activity. TRPV1 is well known for its participation in chemical, thermal and nociceptive sensory transmission. Minocycline antibiotic is considered as an inhibitor of microglial activation therefore it was used for blocking neuroinflammation. The experimental part is comparing an impact of substances applied to the model of tachyphylaxis used for monitoring of nociceptive transmission changes according to decreasing activity of TRPV1 receptors. Electrophysiological recording of miniature excitatory postsynaptic currents from neurons in the Rexed laminae I. and II. of spinal dorsal horn was used. The results of my measurements show that minocycline is able to suppress acute effects of paclitaxel application in vitro if the spinal slice is incubated...
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The role of TRPV1 receptors in chemokine CCL2 induced modulation of nociceptive synaptic transmission at spinal cord level
Adámek, Pavel ; Paleček, Jiří (advisor) ; Krůšek, Jan (referee)
Modulation of nociceptive synaptic transmission in the spinal cord dorsal horn is a significant mechanism in the development and maintenance of different pathological pain states. Accumulating evidence indicates that the TRPV1 (transient receptor potential vanilloid 1) receptor and chemokine CCL2 (C-C motif ligand 2) may play a critical role in this process. The aim of this diploma thesis was to investigate the CCL2 induced modulation of nociceptive synaptic transmission in the dorsal horn of spinal cord and the role of the TRPV1 receptors. To investigate this aim patch-clamp recordings of spontaneous and miniature excitatory postsynaptic currents (sEPSC, mEPSC) from superficial dorsal horn neurons in acute rat lumbar spinal cord slices were used. After acute application of CCL2 on the slice preparation from naïve animals, a frequency increase of both sEPSC and mEPSC was present. This CCL2 induced increase in both sEPSC and mEPSC frequency was prevented by the TRPV1 receptor antagonist SB366791 application. No changes were observed in the amplitudes of sEPSC or mEPSC after application of the CCL2, SB366791, or co-application of CCL2 and SB366791. This suggests that the observed changes were mediated predominantly by presynaptic mechanisms. The preliminary results indicate that after chronic constriction...
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Modulation of synaptic transmission, studies on spinal cord slices in vitro
Mrózková, Petra ; Paleček, Jiří (advisor) ; Krůšek, Jan (referee)
Modulation of a synaptic transmission in the spinal cord dorsal horn plays a key role in nociceptive signalling, especially in states of pathological pain. The goal of this study was to develop a method for calcium imaging in spinal cord slices in vitro. This method allowed us to record changes of intracellular free calcium ions concentration (iCa2+ ), that are a major mediator of neuronal plasticity. In this work, we have focused on application of this method in a conventional fluorescence microscope and on the role of different neuromodulators of synaptic activity. Changes of iCa2+ induced by dorsal root electrical stimulation were recorded altogether in 744 dorsal horn (lamina I and II) neurons. In the first series of experiments, stimulation protocols activating preferentially A and A + C dorsal root fibers were used and long-term stability of the calcium responses was verified. The dorsal root stimulation induced in the neurons fast and delayed type of calcium response. Application of AMPA and NMDA receptors antagonists, CNQX (50μM) and MK801 (45μM), reduced the calcium response amplitude and confirmed the importance of glutamate receptors in synaptic activation. In several experiments the effect of capsaicin a TRPV1 receptors agonist, application was tested. Application of even low...
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Huntington's disease modeling and stem cell therapy in spinal cord disorders and injury
Hruška-Plocháň, Marián ; Motlík, Jan (advisor) ; Bjarkam, Carsten (referee) ; Roth, Jan (referee)
Neurological disorders affect more than 14% of the population worldwide and together with traumatic brain and spinal cord injuries represent major health, public and economic burden of the society. Incidence of inherited and idiopathic neurodegenerative disorders and acute CNS injuries is growing globally while neuroscience society is being challenged by numerous unanswered questions. Therefore, research of the CNS disorders is essential. Since animal models of the CNS diseases and injuries represent the key step in the conversion of the basic research to the clinics, we focused our work on generation of new animal models and on their use in pre-clinical research. We generated and characterized transgenic minipig model of Huntington's disease (HD) which represents the only successful establishment of a transgenic model of HD in minipig which should be valuable for testing of long term safety of HD therapeutics. Next, we crossed the well characterized R6/2 mouse HD model with the gad mouse model which lacks the expression of UCHL1 which led to results that support the theory of "protective" role of mutant huntingtin aggregates and suggest that UCHL1 function(s) may be affected in HD disturbing certain branches of Ubiquitin Proteasome System. Traumatic spinal cord injury and Amyotrophic Lateral...
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Training for autocatheterization in patients with spinal cord lesions from the perspective of an occupational therapist
Šestáková, Eva ; Kulhánková, Zuzana (advisor) ; Čábelková, Alžběta (referee)
This bachelor thesis deals with the rehearsal of self-catheterisation by spinal lesion patients from the occupational therapist's point of view. The aim of the thesis is to create a selfcatheterisation manual for patients after spinal lesion, which might be beneficial for paraplegic as well as tetraplegic patients. The manual contains information about the self- catheterisation procedure and offers instructions, methods and alternatives of compensatory aids and strategies for becoming independent in one of the ADL activities - urinary bladder emptying. The thesis is divided into a theoretical and practical part. The theoretical part clearly explains the problems of tetraplegia and the level of disability and analyses the effects of spinal injury to urinary bladder functioning. The practical part is concerned with the benefits of the Self-catheterisation Rehearsal by Spinal Lesion Patients from the Occupational Therapist's Point of View manual which were examined through a qualitative research. Two case studies including a detailed medical history, the course of therapies and therapy evaluation were created through this research method. The Self-catheterisation Rehearsal by Spinal Lesion Patients from the Occupational Therapist's Point of View manual is to be found in the practical part of this...
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Magnetic resonance volumetric techniques in monitoring disease activity in patients with multiple sclerosis
Andělová, Michaela ; Horáková, Dana (advisor) ; Vymazal, Josef (referee) ; Hluštík, Petr (referee)
Clinical course of multiple sclerosis (MS) is heterogenous and white matter lesion count and volume on brain magnetic resonance imaging (MRI) correlate with clinical course only partially. Therefore, there is an urgent need for more reliable prognostic biomarkers. This work explored three imaging markers - spinal cord (SC) imaging (specifically SC volume measurement and assessment of focal and diffuse SC changes), atlas-based model of "disconnectome" (i.e. disruption of brain connectivity due to white matter lesions) and periventricular white matter gradient assessed with T1 relaxometry. For the SC projects, we assessed MRI from 2044 MS patients with a semi-automatic method for SC volume measurement. We confirmed (i) a relationship between diffuse SC changes, SC volume and disability; (ii) a novel finding was that in patients with EDSS ≤ 4.0, diffuse changes contributed to higher disability more than SC volume; (iii) SC volume explains the paradox in patients with dissociation between brain white matter lesion load and disability; (iv) SC focal and/or diffuse changes are present in 75% patients with early MS, of which 43% have diffuse changes that are related to brainstem lesions. In the disconnectome project, we evaluated the disruption of connectivity caused by white mater lesions in 745 patients...
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Modulation of spinal nociceptive mechanisms under pathological conditions
Mužík, David ; Špicarová, Diana (advisor) ; Smejkalová, Terézia (referee)
Pain is a crucial component of the body's innate defenses, which helps us to respond to the damage that is threatening or imminent. If the pain persists even after the injury has healed, or arises for no apparent reason, it itself becomes harmful. Nociception begins with the detection of a noxious stimulus that irritates free nerve endings on the peripheral projections of spinal ganglion neurons. If the stimulus induces depolarization of the cell and an action potential forms, information of the stimulus is conducted by thinly myelinated Aδ fibers, or unmyelinated C fibers to the spinal cord dorsal horn. Here, the first synapses of sensory pathways are located, which allow the transmission of nociception to secondary afferent neurons, and these further direct the information to the higher centers of the CNS. Synapses in the dorsal horn are key to modulating nociceptive signaling, in which the endocannabinoid system, including endogenous cannabinoids and their receptors, plays a significant role. However, under pathological conditions such as the development of neuropathic pain or neuroinflammation, changes in the expression and function of agonists and receptors of the endocannabinoid system occur. These changes are of great importance in the onset and persistence of pathological pain. The study of...
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Modelovanie ochorenia a štúdium regeneračných procesov v Huntingtonovej chorobe a ALS in vivo
Hruška-Plocháň, Marián
Neurological disorders affect more than 14% of the population worldwide and together with traumatic brain and spinal cord injuries represent major health, public and economic burden of the society. Incidence of inherited and idiopathic neurodegenerative disorders and acute CNS injuries is growing globally while neuroscience society is being challenged by numerous unanswered questions. Therefore, research of the CNS disorders is essential. Since animal models of the CNS diseases and injuries represent the key step in the conversion of the basic research to the clinics, we focused our work on generation of new animal models and on their use in pre-clinical research. We generated and characterized transgenic minipig model of Huntington's disease (HD) which represents the only successful establishment of a transgenic model of HD in minipig which should be valuable for testing of long term safety of HD therapeutics. Next, we crossed the well characterized R6/2 mouse HD model with the gad mouse model which lacks the expression of UCHL1 which led to results that support the theory of "protective" role of mutant huntingtin aggregates and suggest that UCHL1 function(s) may be affected in HD disturbing certain branches of Ubiquitin Proteasome System. Traumatic spinal cord injury and Amyotrophic Lateral...
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