The structure of afferent neurons is pseudounipolar. Studies suggest that they relay action potentials (APs) to both directions of the T-junctions to reach the cell body and the spinal cord. Moreover, the somata are electrically excitable and shown to be able to transmit the signals to associated satellite cells. Our study demonstrates that this transmission can go further and pass onto passive neighbouring somata, if they are in direct contact with same satellite cells. The neurons activate the satellite cells by releasing ATP. This triggers the satellite cells to exocytose acetylcholine to the neighbouring neurons. In addition, the ATP inhibits the nicotinic receptors of the neurons by activating P2Y receptors and initiating the G-protein-mediated pathway, thus reducing the signals that return to the neurons that initiated the signals. This “sandwich synapse” represents a unique pathway by the ectopic release between the somata and the satellite cells.

Neurologically normal individuals show a leftward spatial bias and tend to collide with objects on the right side more frequently than on the left. The upper visual field is associated with extrapersonal space, and mediated by the ventral stream through parvocellular projections. The lower visual field is associated with peripersonal space, and mediated by the dorsal stream through magnocellular projections. Upper and lower visual field differences have been observed in perceptual asymmetries but results have been mixed. Object- and space-based coordinates also both influence the leftward bias; however their relative contributions are unknown as similar spatial conditions are often collapsed across. More left-side collisions emerged on a route following task in the lower visual field and more right-side collisions were seen in the upper visual field (Thomas, Stuckel, Gutwin, & Elias, 2009). Left-handers made more right-side collisions in the central condition, whereas right-handers showed no bias. Leftward biases on the greyscales task were stronger in the lower visual field; however no distance-based differences were observed (Thomas & Elias, 2010). A stronger spatial bias was found on the greyscales task, whereas a stronger object-based bias was found on the object luminosity task (Thomas & Elias, in press). When individual spatial conditions were examined, the image chosen most often was always located in the lower field. Stimulus type and spatial location interacted to determine which coordinate type contributes more strongly to leftward biases. We also found that the leftward bias on the greyscales task was stronger in the lower visual field during prolonged presentation and in the upper visual field during brief presentation. A global motion task was created to preferentially engage magnocellular projections to the dorsal stream. Isoluminant red/green and blue/yellow colour tasks, which preferentially engage parvocellular projections to the ventral stream, were also created. Leftward biases were seen on the greyscales and motion tasks. On an isoluminant colour task, biases were significantly weakened, suggesting leftward biases exhibited by neurologically normal people are mediated by magnocellular projections to the dorsal stream and this preferential processing leads to a lower visual field advantage on the greyscales task.

Peripheral nerve injury induces a robust regenerative state in sensory neurons that includes elevated expression of injury/regeneration-associated genes. The molecular signal(s) underlying the transition to the regenerating state are largely unknown. Brain-derived neurotrophic factor (BDNF) is the sole identified neurotrophin that is upregulated in sensory neurons following peripheral nerve injury. As members of the neurotrophin family exert a profound influence on the intact phenotype of sensory neurons, I hypothesize that injury-associated alterations in BDNF expression play a similar role in the injured/regenerating response. Antagonizing endogenous BDNF with a function-blocking antibody prevented increases in injury/regeneration-associated gene expression and decreased the growth capabilities of the injured sensory neurons. However, BDNF was not important for maintaining this cell body response in injured neurons. The elevation of BDNF expression in injured sensory neurons either through intrathecal infusion or electrical stimulation was associated with increased injury/regeneration-associated gene expression in a dose dependent manner and the latter corresponded to increased sensory axonal regeneration. Though BDNF was able to induce and enhance the intrinsic cell body response of injured sensory neurons, exogenous BDNF was not sufficient to induce an injury phenotype in intact sensory neurons. Thus, additional signals are likely induced by the injury response. In conclusion, BDNF plays a critical role in inducing the regenerative state in sensory neurons following injury and strategies aimed at elevating levels of BDNF available to the injured sensory neuron during the inductive phase improve the cell body response.

Peripheral nerve injury induces a robust regenerative state in sensory neurons that includes elevated expression of injury/regeneration-associated genes. The molecular signal(s) underlying the transition to the regenerating state are largely unknown. Brain-derived neurotrophic factor (BDNF) is the sole identified neurotrophin that is upregulated in sensory neurons following peripheral nerve injury. As members of the neurotrophin family exert a profound influence on the intact phenotype of sensory neurons, I hypothesize that injury-associated alterations in BDNF expression play a similar role in the injured/regenerating response. Antagonizing endogenous BDNF with a function-blocking antibody prevented increases in injury/regeneration-associated gene expression and decreased the growth capabilities of the injured sensory neurons. However, BDNF was not important for maintaining this cell body response in injured neurons. The elevation of BDNF expression in injured sensory neurons either through intrathecal infusion or electrical stimulation was associated with increased injury/regeneration-associated gene expression in a dose dependent manner and the latter corresponded to increased sensory axonal regeneration. Though BDNF was able to induce and enhance the intrinsic cell body response of injured sensory neurons, exogenous BDNF was not sufficient to induce an injury phenotype in intact sensory neurons. Thus, additional signals are likely induced by the injury response. In conclusion, BDNF plays a critical role in inducing the regenerative state in sensory neurons following injury and strategies aimed at elevating levels of BDNF available to the injured sensory neuron during the inductive phase improve the cell body response.

Neurologically normal individuals show a leftward spatial bias and tend to collide with objects on the right side more frequently than on the left. The upper visual field is associated with extrapersonal space, and mediated by the ventral stream through parvocellular projections. The lower visual field is associated with peripersonal space, and mediated by the dorsal stream through magnocellular projections. Upper and lower visual field differences have been observed in perceptual asymmetries but results have been mixed. Object- and space-based coordinates also both influence the leftward bias; however their relative contributions are unknown as similar spatial conditions are often collapsed across. More left-side collisions emerged on a route following task in the lower visual field and more right-side collisions were seen in the upper visual field (Thomas, Stuckel, Gutwin, & Elias, 2009). Left-handers made more right-side collisions in the central condition, whereas right-handers showed no bias. Leftward biases on the greyscales task were stronger in the lower visual field; however no distance-based differences were observed (Thomas & Elias, 2010). A stronger spatial bias was found on the greyscales task, whereas a stronger object-based bias was found on the object luminosity task (Thomas & Elias, in press). When individual spatial conditions were examined, the image chosen most often was always located in the lower field. Stimulus type and spatial location interacted to determine which coordinate type contributes more strongly to leftward biases. We also found that the leftward bias on the greyscales task was stronger in the lower visual field during prolonged presentation and in the upper visual field during brief presentation. A global motion task was created to preferentially engage magnocellular projections to the dorsal stream. Isoluminant red/green and blue/yellow colour tasks, which preferentially engage parvocellular projections to the ventral stream, were also created. Leftward biases were seen on the greyscales and motion tasks. On an isoluminant colour task, biases were significantly weakened, suggesting leftward biases exhibited by neurologically normal people are mediated by magnocellular projections to the dorsal stream and this preferential processing leads to a lower visual field advantage on the greyscales task.

<p>Spinal cord injury (SCI) and other neurological diseases and disorders can cause urinary dysfunction that can cause serious health problems and reduce an individual's quality of life. Current methods for treating urinary dysfunction have major limitations or provide inadequate improvement in urinary symptoms. Pudendal nerve stimulation is a potential means of restoring control of bladder function in persons with neurological disease or spinal cord injury. Bladder contraction and relaxation can be evoked by pudendal afferent stimulation, and peripheral pudendal afferent branches may be ideal targets for a bladder control neural prosthesis. This dissertation investigates control of bladder function by selective activation of pudendal afferents.</p> <p>This study investigated the ability to improve both urinary continence and micturition by both direct and minimally-invasive electrical stimulation of selected pudendal afferents in &#945;-chloralose anesthetized male cats. Direct stimulation of the pudendal afferents in the dorsal nerve of the penis (DNP), percutaneous DNP stimulation, and intraurethral stimulation were used to investigate the bladder response to selective activation of pudendal afferents. Finite element modeling of the cat lower urinary tract was used to investigate the impact of intraurethral stimulation location and intraurethral electrode configuration on activation of pudendal afferents. Also, the impact of pharmacological and surgical block of sympathetic activity to the bladder on the bladder reflexes evoked by DNP stimulation was investigated to determine the role of the sympathetic bladder innervation on the mechanism of bladder activation by pudendal afferent stimulation.</p> <p>The DNP is an ideal target for restoring urinary function because stimulation at low frequencies (5-10 Hz) improves urinary continence, while stimulation at high frequencies (33-40 Hz) improves urinary voiding. Intraurethral stimulation is a valid method for clinical investigation of the ability to evoke bladder inhibition and activation via selective activation of the DNP or cranial sensory branch (CSN) of the pudendal nerve. In the cat, intraurethral stimulation can activate the bladder via two distinct neural pathways, a supraspinal pathway reflex activated by the CSN and a spinal reflex activated by the DNP. Finite element modeling revealed the importance of urethral location for selective pudendal afferent activation by intraurethral stimulation. Finally, the sympathetic bladder pathway does not play a significant role in the mechanism mediating bladder activation by DNP stimulation. These findings imply that selective pudendal afferent stimulation is a promising approach for restoring control of bladder function to individuals with SCI or other neurological disorders.</p> / Dissertation

Η μελέτη της φυλοσύνδετης μετάλλαξης wisertsl (1-21.7, 7E) της Drosophila melanogaster αποκάλυψε ότι υπεύθυνο για τους φαινότυπους φαγωμένα/τσιμπημένα φτερά, ελαφρώς ανώμαλα μάτια και τη θερμοευαισθησία είναι το γονίδιο CG32711, που ονομάσαμε wiser (wings scalloped-eyes rough). Το γονίδιο wiser είναι απαραίτητο για τη σωστή ανάπτυξη της Drosophila melanogaster. Η μετάλλαξη wisertsl χαρτογραφείται στη 5΄ ρυθμιστική περιοχή του γονιδίου wiser. Στην ίδια περιοχή χαρτογραφείται και η θανατογόνος μετάλλαξη wiserPL26. Παραπέρα μελέτη του γονιδίου wiser με τη χρήση αυτών των δυο μεταλλάξεων και του διαγονιδίου UAS-wiser αποκάλυψε ότι: α) Οι μεταλλάξεις wisertsl και wiserPL26 ενισχύουν το φαινότυπο των μεταλλάξεων Beadex1 και Serrate1. Το γονίδιο wiser αλληλεπιδρά με τα γονίδια Beadex και Serrate, τα οποία εμπλέκονται στην ενεργοποίηση του Notch μονοπατιού σηματοδότησης κατά μήκος του ραχιοκοιλιακού άξονα του φτερού. Η παρατήρηση αυτή δείχνει, ότι το wiser εμπλέκεται στον προσδιορισμό του ραχιοκοιλιακού άξονα. β) Η μετάλλαξη wisertsl σε ομοζυγωτία μειώνει σημαντικά την έκφραση των διαγονιδίων fringe-lacZ, m8-lacZ, wingless-lacZ, vestigial-lacZ και Distalless-lacZ, αλλάζει το πρότυπο έκφρασης του mβ-lacZ και δεν επηρεάζει την έκφραση του apterous-lacZ στους εμβρυικούς δίσκους του φτερού προνυμφών 3ου σταδίου. Τα αποτελέσματα αυτά έδειξαν, ότι το γονίδιο wiser δρα μετά το γονίδιο apterous και πριν το γονίδιο fringe, που είναι τροποποιητής του υποδοχέα Notch. Επομένως, η δράση του Notch εξαρτάται και από το wiser. γ) Εκτοπική έκφραση του διαγονιδίου UAS-wiser με οδηγό το ap-Gal4, έδειξε ότι διασώζει μερικώς το φαινότυπο apterous- αλλά όχι το φαινότυπο Serrate1. δ) Εκτοπική έκφραση του UAS-wiser με οδηγό το dpp-Gal4, επηρεάζει την έκφραση του wingless-lacZ αλλά όχι των apterous, fringe, mβ, m8, vestigial και Distalless στους εμβρυικούς δίσκους φτερού. ε) Η δημιουργία μιτωτικών κλώνων με το σύστημα FRT/FLP, σε θηλυκά άτομα wiserPL26/+ οδήγησε στη δημιουργία κλώνων +/+ και wiserPL26/wiserPL26 διαφορετικού μέγεθος στους εμβρυικούς δίσκους του φτερού. Οι πρώτοι (+/+), έχουν σημαντικά μεγαλύτερο μέγεθος από τους δεύτερους όταν συμβαίνουν στη περιοχή του εμβρυικού δίσκου που θα δώσει το φτερό του ακμαίου ατόμου. Στα ακμαία θηλυκά οι σωματικοί κλώνοι εκδηλώνονται με το φαινότυπο φαγωμένα φτερά. Σωματικοί κλώνοι παρατηρήθηκαν και στα μάτια των ακμαίων. Τα αποτελέσματα των μιτωτικών κλώνων δείχνουν ότι το γονίδιο wiser εμπλέκεται στον πολλαπλασιασμό των κυττάρων. Όλα τα παραπάνω αποτελέσματα, δείχνουν ότι το γονίδιο wiser είναι απαραίτητο για την ανάπτυξη του φτερού, καθώς εμπλέκεται στο σχηματισμό του ραχιοκοιλιακού άξονα και επηρεάζει τον πολλαπλασιασμό των κυττάρων. / The analysis of the X-linked wisertsl (1-21.7, 7E) mutation in Drosophila melanogaster has shown that responsible for the scalloped phenotype and the temperature sensitivity is the CG32711 gene, which we name wiser (wings scalloped-eyes rough). The gene wiser is essential for Drosophila development. The wisertsl mutation is mapped at the 5′ regulatory region of the gene CG32711. The wiserPL26 lethal mutation is mapped in the same region. Using these two mutations and a UAS-wiser transgene we have shown that: a) The wisertsl and wiserPL26 mutations increase the wing scalloping (phenotype) of the mutations Beadex1 and Serrate1. The genes Beadex and Serrate are implicated in the activation of Notch signaling pathway along the dorsal-ventral axis of the wing. This observation indicates that the wiser gene is involved in determination of dorsal-ventral axis. b) The wisertsl mutation in homozygous condition reduces substantially the expression of fringe-lacZ, m8-lacZ, wingless-lacZ, vestigial-lacZ and Distalless-lacZ transgenes, alters the expression pattern of mβ-lacZ and does not affect the expression of apterous-lacZ transgene in the wing imaginal disc. This indicates that the expression of fringe (a modifier of Notch receptor) is regulated by wiser too. c) Ectopic expression of UAS-wiser by the ap-Gal4 driver partially rescues apterous- but not Serrate1 phenotype. d) Ectopic expression of UAS-wiser by the dpp-Gal4 driver affects the expression of wingless and does not affects the expression of apterous, fringe, mβ, m8, vestigial and Distalless in the wing imaginal disc (revealed by the corresponding -lacZ strains). e) Induction of somatic clones with the FRT/FLP system in wiserPL26/+ mutants led to mitotic +/+ and wiserPL26/wiserPL26 clones of different sizes. The first clones were much larger than the second ones in the territory of wing pouch. Adult females with scalloped wings were also produced. These results indicate that the wiser gene is involved in cell proliferation. All the above findings suggest that the wiser gene is essential for wing development and cell proliferation.

Luman (CREB3, LZIP) is a basic leucine zipper transcription factor involved in regulation of the unfolded protein response (UPR), dendritic cell maturation, and cell migration. But despite reported expression in primary sensory neurons, little is known about its role in the nervous system. Luman mRNA from rat sensory neurons was amplified and its coding sequence was determined. The rat Luman cDNA contains a full-length open reading frame encoding 387 amino acids, and the recombinant protein generated from this clone activated transcription from UPR elements. Quantitative RT-PCR revealed rat Luman transcripts in a variety of rat tissues with the highest levels in nervous system tissue. In situ hybridization confirmed the findings and demonstrated that the Luman mRNA hybridization signal localizes to neurons and satellite glial cells in dorsal root ganglia (DRG), the cytoplasm of hepatocytes in liver, and the hippocampal pyramidal cell layers in CA1 and CA3 and the granular cell layer of the dentate gyrus. Luman protein localizes with axonal endoplasmic reticulum (ER) components along the axon length within the sciatic nerve and is activated by sciatic nerve injury. Adult sensory axons also contain Luman mRNA which is translated within the axon and transported to the cell body via the importin-mediated retrograde transport system in response to nerve injury. Further, creation of an N-terminal, C-terminal dual fluorescence-tagged Luman adenoviral construct allowed visualization of the cleavage and retrograde translocation of the N-terminal portion of Luman to the nucleus in real time in vivo and in vitro. Neuronal or subcellular axonal knockdown of Luman significantly impaired the intrinsic ability of injury-conditioned, but not naïve, sensory neurons to extend the regeneration-associated elongating form of neurites. Sciatic nerve crush injury also induced activation of the UPR in axotomized DRGs, including genes linked to cholesterol biosynthesis. Knockdown of Luman decreased the activation of UPR and cholesterol biosynthesis, and axotomy-inducted increases in neurite outgrowth, which could be largely rescued with either mild UPR inducer treatment or cholesterol supplementation. Together these findings provide novel insights linking remote injury-associated axonal ER responses to the regenerative growth capacity of adult sensory neurons via axonal activation and synthesis of Luman and reveal a role for the UPR in regulation of axotomy-induced neurite outgrowth that is critically dependent on Luman.

Microvascular permeability is a pharmacologic indicator of tumor response to therapy, and it is expected that this biomarker will evolve into a clinical surrogate endpoint and be integrated into protocols for determining patient response to antiangiogenic or antivascular therapies. The goal of this research is to develop a method by which microvascular permeability (Ktrans) and vascular volume (vp) as measured by DCE-MRI were directly compared to the same parameters measured by intravital fluorescence microscopy in an MRI-compatible window chamber model. Dynamic contrast enhanced-MRI (DCE-MRI) is a non-invasive, clinically useful imaging approach that has been used extensively to measure active changes in tumor microvascular hemodynamics. However, uncertainties exist in DCE-MRI as it does not interrogate the contrast reagent (CR) itself, but the effect of the CR on tissue water relaxivity. Thus, direct comparison of DCE-MRI with a more quantitative measure would help better define the derived parameters. The combined imaging system was able to obtain both dynamic contrast-enhanced MRI data high spatio-termporal resolution fluorescence data following injection of fluorescent and gadolinium co-labeled albumin. This approach allowed for the cross-validation of vascular permeability data, in relation tumor growth, angiogenesis and response to therapy in both imaging systems.

No description available.