Diss. (sammanfattning) Stockholm : Karol. inst. / Härtill 7 uppsatser.

Sun Qiang. / "December 1999." / Thesis (Ph.D.)--Chinese University of Hong Kong, 1999. / Includes bibliographical references (p. 119-139). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web.

The activity of glutamate transporters is essential for the temporal and spatial regulation of the neurotransmitter concentration in the synaptic cleft which is critical for proper neuronal signaling. Because of their role in controlling extracellular glutamate concentrations, dysfunctional glutamate transporters have been implicated in several neurodegenerative diseases and psychiatric disorders. Therefore, investigating the mechanism of substrate transport by these transporters is essential in understanding their behavior when they malfunction. A bacterial glutamate transporter homologue has been successfully crystallized revealing the molecular architecture of glutamate transporters. However, many important questions remain unanswered. In this thesis, I will address the role of D439 in the binding of Na+, and I will identify other electrogenic steps that contribute to the total electrogenicity of the transporter cycle. The role of D439 in the binding of Na+ to the transporter was explored previously in this lab. While it was proposed that the effect of D439 in Na+ binding is indirect, the results described in this thesis provides added support to this work. Here, I will show that the D439 mutation changed the pharmacology of EAAC1 such that THA was converted from a transported substrate to a competitive inhibitor. I will also show that Na+ binding to the substrate-bound mutant transporter occurred with the same affinity as that of Na+ to the substrate-bound wild-type transporter. Therefore, based on these results, D439 is not directly involved in the binding of Na+ to the substrate-bound transporter, but that its effect is rather indirect through changing the substrate binding properties. Na+ binding steps to the empty transporter and to the glutamate-bound EAAC1 contribute only 20% of the total electrogenicity of the glutamate transporter reactions cycle. While K+-induced relocation has been proposed to be electrogenic, there is no experimental evidence that supports it. In this work, I will show that the K+-induced relocation of the empty transporter is electrogenic. Moreover, the results in this work show that the K+-dependent steps are slower than the steps associated with the Na+/glutamate translocation suggesting that the K+-induced relocation determines the transporter?s properties at steady state.

Glutamate, the predominant excitatory neurotransmitter in the central nervous system, is involved in almost all aspects of neurological function including cognition, motor function, memory, learning, decision making, and neuronal plasticity. For normal neurological function, glutamate signaling must be properly regulated. Disrupted glutamate regulation plays a pivotal role in the acute pathophysiology of traumatic brain injury (TBI), disrupting neuronal signaling, initiating secondary injury cascades, and producing excitotoxicity. Increases in extracellular glutamate have been correlated with unfavorable outcomes in TBI survivors, emphasizing the importance of glutamate regulation. The aim of this thesis was to examine disruptions in the regulation of extracellular glutamate after experimental TBI. In these studies, we used glutamate-sensitive microelectrode arrays (MEAs) to examine the regulation of extracellular glutamate two days after diffuse brain injury. First, we examined which brain regions were vulnerable to post-traumatic increases in extracellular glutamate. We detected significant increases in extracellular glutamate in the dentate gyrus and striatum, which correlated to the severity of brain injury. Second, we examined the regulation of extracellular glutamate by neurons and glia to determine the mechanisms responsible for post-traumatic increases in extracellular glutamate. In the striatum of brain-injured rats, we detected significant disruptions in release of glutamate by neurons and significant decreases in the removal of glutamate from the extracellular space by glia. Third, we examined if a novel therapeutic strategy, a viral-vector mediated gene delivery approach, could improve the regulation of extracellular glutamate. Infusion of an adeno-associated virus expressing a glutamate transporter into the rat striatum produced significant improvements in glutamate clearance, identifying a novel strategy to reduce excitotoxicity. Lastly, we examined the translational potential of MEAs as novel neuromonitoring device for clinical TBI research. Overall, these studies have demonstrated the translational potential of MEAs to aid in the diagnosis and treatment of TBI survivors.

The underlying nociceptive mechanisms in temporomandibular joint (TMJ) and masticatory muscles in many pain conditions are still unclear, largely due to the limited study of peripheral and central neural mechanisms affecting craniofacial musculoskeletal tissues. This study provided evidence in support of Hypothesis 1: Peripheral glutamatergic and capsaicin-sensitive mechanisms modulate the properties of primary afferents and brainstem neurons processing deep craniofacial nociceptive information. Effects of glutamate and capsaicin injected into the receptive field of deep craniofacial nociceptive afferents or TMJ of TMJ-responsive nociceptive neurons in trigeminal subnucleus caudalis/upper cervical cord (Vc/UCC) were studied in halothane-anesthetized rats. When injected alone, glutamate and capsaicin activated and induced peripheral sensitization in many afferents. Following glutamate injection, capsaicin-evoked activity was greater than that evoked by capsaicin alone, whereas following capsaicin injection, glutamate-evoked responses were similar to those of glutamate alone. When injected alone, glutamate and capsaicin also activated and induced central sensitization in most Vc/UCC neurons. Following glutamate injection, capsaicin evoked greater activity and less sensitization compared with capsaicin alone, whereas following capsaicin, glutamate was less effective in activating and sensitizing most Vc/UCC neurons. This apparent desensitizing effect of capsaicin on glutamate-evoked excitability of Vc/UCC neurons contrasts with the lack of capsaicin-induced modulation of glutamate-evoked afferent excitability, suggesting that peripheral and central sensitization may be differentially involved in the nociceptive effects of glutamate and capsaicin applied to deep craniofacial tissues. Further evidence of glutamate-capsaicin interactions was documented in the attenuation by TMJ pre-injection of glutamate receptor antagonists of jaw muscle activity reflexly evoked by TMJ injection of capsaicin. Moreover, additional findings support Hypothesis 2: Surgical cutaneous incision modulates the properties of brainstem neurons processing deep craniofacial nociceptive information. TMJ-responsive nociceptive Vc/UCC neurons could be activated by surgical incision of the skin overlying the TMJ and this incision-induced afferent barrage caused nociceptive neurons to be temporarily refractory to further capsaicin-induced central sensitization. These novel findings suggest that peripheral glutamate and capsaicin receptor mechanisms as well as surgical cutaneous incision may be involved in the nociceptive processing of deep craniofacial afferent inputs and may interact to modulate both activation as well as sensitization evoked from these tissues.

The underlying nociceptive mechanisms in temporomandibular joint (TMJ) and masticatory muscles in many pain conditions are still unclear, largely due to the limited study of peripheral and central neural mechanisms affecting craniofacial musculoskeletal tissues. This study provided evidence in support of Hypothesis 1: Peripheral glutamatergic and capsaicin-sensitive mechanisms modulate the properties of primary afferents and brainstem neurons processing deep craniofacial nociceptive information. Effects of glutamate and capsaicin injected into the receptive field of deep craniofacial nociceptive afferents or TMJ of TMJ-responsive nociceptive neurons in trigeminal subnucleus caudalis/upper cervical cord (Vc/UCC) were studied in halothane-anesthetized rats. When injected alone, glutamate and capsaicin activated and induced peripheral sensitization in many afferents. Following glutamate injection, capsaicin-evoked activity was greater than that evoked by capsaicin alone, whereas following capsaicin injection, glutamate-evoked responses were similar to those of glutamate alone. When injected alone, glutamate and capsaicin also activated and induced central sensitization in most Vc/UCC neurons. Following glutamate injection, capsaicin evoked greater activity and less sensitization compared with capsaicin alone, whereas following capsaicin, glutamate was less effective in activating and sensitizing most Vc/UCC neurons. This apparent desensitizing effect of capsaicin on glutamate-evoked excitability of Vc/UCC neurons contrasts with the lack of capsaicin-induced modulation of glutamate-evoked afferent excitability, suggesting that peripheral and central sensitization may be differentially involved in the nociceptive effects of glutamate and capsaicin applied to deep craniofacial tissues. Further evidence of glutamate-capsaicin interactions was documented in the attenuation by TMJ pre-injection of glutamate receptor antagonists of jaw muscle activity reflexly evoked by TMJ injection of capsaicin. Moreover, additional findings support Hypothesis 2: Surgical cutaneous incision modulates the properties of brainstem neurons processing deep craniofacial nociceptive information. TMJ-responsive nociceptive Vc/UCC neurons could be activated by surgical incision of the skin overlying the TMJ and this incision-induced afferent barrage caused nociceptive neurons to be temporarily refractory to further capsaicin-induced central sensitization. These novel findings suggest that peripheral glutamate and capsaicin receptor mechanisms as well as surgical cutaneous incision may be involved in the nociceptive processing of deep craniofacial afferent inputs and may interact to modulate both activation as well as sensitization evoked from these tissues.

Thesis (Ph. D.)--University of Alabama at Birmingham, 2007. / Title from first page of PDF file (viewed Feb. 7, 2008). Includes bibliographical references.

Diss. (sammanfattning) Stockholm : Karol. inst., 2004. / Härtill 5 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2003. / Härtill 7 uppsatser.

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2010.