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Exploring the role of opioid signaling in modulation of microglial function
Mali, Akash Shivling ; Novotný, Jiří (advisor) ; Svoboda, Petr (referee) ; Machová Urdzíková, Lucia (referee)
Microglial activation is the most important component of neuroinflammation. It appears that opioids may affect microglial M1/M2 polarization in different ways depending on the type of receptor employed. In addition to opioid receptors, Toll-like receptor 4 (TLR4) of the innate immune system can also be activated by some opioid ligands and thus elicit specific cellular responses. Although opioid receptors (ORs) are known to regulate neurotransmission in various peptidergic neurons, their potential role in modulation of microglial function remains largely unknown. In this study, we investigated the effects of OR agonists, namely DAMGO, DADLE, and U-50488, on polarization and metabolic modulation of C8-B4 microglial cells. Our findings have revealed that opioids effectively suppress lipopolysaccharide (LPS)-triggered M1 polarization and promote the M2 polarization state. This was evidenced by decreased phagocytic activity, decreased production of nitric oxide (NO), diminished expression of proinflammatory cytokines such as TNF-α, IL-1β, IL-6, IL-86, and IL-12 beta p40, along with an increased migration rate and elevated expression of anti-inflammatory markers such as IL-4, IL-10, IL-13 arginase 1, and CD206 in microglia compared to cells influenced by LPS. Furthermore, we have demonstrated that...
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Regeneration and structural changes of the nerve tissue after the extracellular matrix modification
Vallová, Jana ; Machová Urdzíková, Lucia (advisor) ; Hampl, Aleš (referee) ; Hubálek Kalbáčová, Marie (referee)
Regeneration and structural changes of the nerve tissue after the extracellular matrix modification. Abstract ECM modification may contribute to changes in nerve tissue plasticity. Therefore, 4- methylubulliferone is used in our study, which disrupted the structure of the perineuronal networks surrounding some types of neurons responsible for the formation of memory traces. Mice were fed a 4-MU diet (6.7 mg / g / day) for 6 months, which improved their memory skills in a spontaneous recognition test without a significant adverse effect on the kidneys, liver, or joints.In the next part of the study, an extracellular matrix (UC-ECM) was derived from fetal human umbilical cord tissue also generated as biomimetic hydrogel. Due to the generated UC-ECM's low stability and rapid degradation, the structure was stabilized by covalent genipin bonding. Stabilization with 1 mM genipine increased the biological stability of the material. UC-ECM as well as ECM/G didn't show toxicity in vitro in mesenchymal stem cell proliferation; axonal budding or neural stem cell growth and differentiation were not adversely affected. The biocompatibility of both materials was verified in vivo by applying the material to an intracortical photothrombotic rat lesion, where gelation and infiltration of the lesion and hydrogel by host...
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Experimental model of the ventral spinal cord lesion in rats.
Štěpánková, Kateřina ; Machová Urdzíková, Lucia (advisor) ; Martončíková, Marcela (referee)
A spinal cord injury (SCI) is a damage to the spinal cord that causes permanent or temporary changes in motor and sensory functions. In humans, the traumatic impact to spinal cord is mostly directed from the ventral part of the spinal column, even though SCI models are principally directed from the dorsal part of the spinal column due to easier surgery. The aim of the work described in this thesis was to develop clinically more relevant, easily reproducible and relatively inexpensive model of the ventral spinal cord lesion in rats which replicates SCI in humans as closely as possible by its pathology, completeness, level and regeneration. For the surgery we used a modification of a balloon- compression technique. The balloon of the 2F embolectomy Fogarty's catheter was placed to the anterior epidural space via laminectomy at the level of T10 and when the final position of the catheter was achieved at the spinal level T8 in front of the anterior median fissure, a balloon was rapidly inflated with 10 μl or 15 μl of water for 5 minutes. Other two groups - laminectomy only and ventral placement of the catheter without inflation were used as controls. The motor functions were evaluated by the BBB test and ladder walking test. In BBB test we observed significantly impaired motor functions in the 15 μl...
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Experimental rat models in traumatic spinal cord injury research
Kloudová, Anna ; Machová Urdzíková, Lucia (advisor) ; Mrózková, Petra (referee)
A large number of spinal injuries that cause damage to the spinal cord occur every year. Such damage to spinal cord tissue leads to a number of dysfunctions that significantly reduce quality of life and therefore the development of effective therapies remains a very important issue. The use of animal models is considered the major tool of this research field, enabling the testing of new treatments as well as observing ongoing pathophysiological processes. Rats are the most frequently used animal model due to the many possible ways of creating a spinal cord lesion on them. The contusion model uses a computer-controlled device to simulate the spinal cord contusion and therefore is highly precise. However, a compression model seems to be the most suitable model for pathophysiological studying. Besides contusion it enables persistent compression to the spinal cord. Transection and hemisection methods are intended for axonal regeneration research, as only these models can reliably distinguish new growing fibers from the preserved ones. Less often photochemical and excitotoxic models are used. Lastly, dislocation and distraction of vertebrae or damage to the spinal cord using heat or cold can be carried out. Each of the models have their own unique features that make them optimal to use in different...
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The Role of Aquaporin 4 channels and Transient Receptor Potential Vanilloid 4 channels in astrocytic swelling
Heřmanová, Zuzana ; Anděrová, Miroslava (advisor) ; Machová Urdzíková, Lucia (referee)
Astrocytes posses a wide range of functions within the brain. In response to ischemic conditions they swell due to increased uptake of osmolytes and they are mainly responsible for cytotoxic edema formation. However, they are also able to regulate their volume by releasing osmolytes together with water via the process of regulatory volume decrease (RVD). The Aquaporin 4 (AQP4) channel and Transient receptor potential vanilloid 4 (TRPV4) channel are suspected to be strongly involved in these processes of astrocytic volume regulation. The goal of the present diploma thesis was to clarify the role of both channels in astrocytic swelling in situ. For our experiments we used a subpopulation of green fluorescent protein-labelled astrocytes from AQP4-deficient (AQP4-/- ), TRPV4-deficient (TRPV4-/- ) and control (Ctrl) mice. Cell volume alterations were induced in acute brain slices by hypoosmotic stress or by oxygen-glucose deprivation (OGD). Data were quantified using fluorescence intensity-based approach in the whole cells and in astrocytic endfeet. Our results indicate, that there is no difference in astrocytic swelling or cell volume recovery between astrocytes from AQP4-/- , TRPV4-/- and control mice when exposed to hypoosmotic stress. On the contrary, volume changes induced by OGD varied...
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Behavioral testing of animals after experimental spinal cord injury.
Freislebenová, Hana ; Machová Urdzíková, Lucia (advisor) ; Kriška, Ján (referee)
Spinal cord injury is one of the most serious injury that leads to a decrease in the quality of life both physically and mentally. Although there is currently no effective treatment, great efforts are being made to develop new treatments. The effectiveness of experimental treatment is verified by behavioral tests performed on animal models, where rats are the most suitable ones for this type of injury. Behavioral tests are divided into several groups depending on what the testing is focused on. Locomotor tests examine the function of a locomotive apparatus, motor tests are used to determine the function of skeletal muscles not primarily involved in locomotion, sensory and motor tests test the functionality of sensory and motor apparatus and their interconnection, sensory tests are designed to test responses to sensory stimuli. Electrophysiological tests and functional magnetic resonance imaging are part of a series of special tests that use artificially induced electrical stimuli to measure muscle activity. This thesis is focused on detailed description of selected behavioral tests, their functionality and comparison of their advantages and disadvantages.
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Curcumin in the treatment of traumatic spinal cord injury
Kloudová, Anna ; Machová Urdzíková, Lucia (advisor) ; Martončíková, Marcela (referee)
Spinal cord injury is a very significant clinical as well as social problem with extensive consequences, affecting the patient and also his/her family. Great efforts have been devoted to searching for an effective treatment, which would improve their situation. This thesis evaluated the effects of the natural compound curcumin on spinal cord injury using an experimental balloon compression model. Male Wistar rats were randomized into two groups following the lesion induction, namely vehicle- or curcumin-treated. The behavioral recovery was evaluated using a set of locomotor and sensory tests and a histological and imunohistochemical analysis was performed. The qPCR method was used to observe the expression of some genes related to regeneration and immune response. It was demonstrated that curcumin improved locomotor recovery after the spinal cord injury, particularly in the early stages. Morphometric analysis of the gray and white matter sparing didn't confirm any differences between the two groups. Nevertheless, the glial scar formation was significantly reduced around the central part of the lesion in the curcumin treated group and also the NF-κB activity was substantially inhibited. The gene expression analysis demontrated downregulation of Gfap and Rantes genes and upregulation of the Irf5 gene...
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Experimental model of the ventral spinal cord lesion in rats.
Štěpánková, Kateřina ; Machová Urdzíková, Lucia (advisor) ; Martončíková, Marcela (referee)
A spinal cord injury (SCI) is a damage to the spinal cord that causes permanent or temporary changes in motor and sensory functions. In humans, the traumatic impact to spinal cord is mostly directed from the ventral part of the spinal column, even though SCI models are principally directed from the dorsal part of the spinal column due to easier surgery. The aim of the work described in this thesis was to develop clinically more relevant, easily reproducible and relatively inexpensive model of the ventral spinal cord lesion in rats which replicates SCI in humans as closely as possible by its pathology, completeness, level and regeneration. For the surgery we used a modification of a balloon- compression technique. The balloon of the 2F embolectomy Fogarty's catheter was placed to the anterior epidural space via laminectomy at the level of T10 and when the final position of the catheter was achieved at the spinal level T8 in front of the anterior median fissure, a balloon was rapidly inflated with 10 μl or 15 μl of water for 5 minutes. Other two groups - laminectomy only and ventral placement of the catheter without inflation were used as controls. The motor functions were evaluated by the BBB test and ladder walking test. In BBB test we observed significantly impaired motor functions in the 15 μl...
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The Role of Aquaporin 4 channels and Transient Receptor Potential Vanilloid 4 channels in astrocytic swelling
Heřmanová, Zuzana ; Anděrová, Miroslava (advisor) ; Machová Urdzíková, Lucia (referee)
Astrocytes posses a wide range of functions within the brain. In response to ischemic conditions they swell due to increased uptake of osmolytes and they are mainly responsible for cytotoxic edema formation. However, they are also able to regulate their volume by releasing osmolytes together with water via the process of regulatory volume decrease (RVD). The Aquaporin 4 (AQP4) channel and Transient receptor potential vanilloid 4 (TRPV4) channel are suspected to be strongly involved in these processes of astrocytic volume regulation. The goal of the present diploma thesis was to clarify the role of both channels in astrocytic swelling in situ. For our experiments we used a subpopulation of green fluorescent protein-labelled astrocytes from AQP4-deficient (AQP4-/- ), TRPV4-deficient (TRPV4-/- ) and control (Ctrl) mice. Cell volume alterations were induced in acute brain slices by hypoosmotic stress or by oxygen-glucose deprivation (OGD). Data were quantified using fluorescence intensity-based approach in the whole cells and in astrocytic endfeet. Our results indicate, that there is no difference in astrocytic swelling or cell volume recovery between astrocytes from AQP4-/- , TRPV4-/- and control mice when exposed to hypoosmotic stress. On the contrary, volume changes induced by OGD varied...
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Neurophysiology of the urinary system after spinal cord injury.
Štěpánková, Kateřina ; Machová Urdzíková, Lucia (advisor) ; Horníková, Daniela (referee)
Spinal cord injury (SCI) is a devastating condition that is a result of trauma generated by traffic accident, violence or falling. However, SCI may not only be caused by trauma, but also by other diseases such as - virus infection, tumors etc.. Any SCI affects the ability of the spinal cord to transmit and receive information and their further processesing. The spinal cord injury itself is followed by other complications that seriously impair the quality of life of the patients. One of the main problems is the dysfunction of the urinary system, which then does not work properly, thereby make patient's life dificult on the health and social side. The aim of this thesis is to summarize pathophysiological consequences of SCI on urinary tract dysfunction. Keywords: neurophysiology, spinal cord injury, paraplegia, urinary tract
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