Neurotrophins BDNF and NT4 influence the development of the rodent gustatory system. Despite binding to the same receptor, TrkB, they have different roles. BDNF is chemo-attractive for gustatory neurons and regulates gustatory neuron targeting and number during development. NT4 regulates gustatory neuron number earlier in development than BDNF, but it is not chemo-attractive and does not regulate gustatory neuron targeting. To elucidate the mechanisms that regulate these processes we have examined which TrkB intracellular signalling pathways are required for specific aspects of gustatory development by studying the effect of specific point mutations in TrkB docking sites. We found that the TrkB/Shc docking site is involved in regulating the survival of geniculate ganglion neurons as a point mutation in this adaptor site (TrkbS/S) caused large losses of these neurons as early as E12.5. These losses were exacerbated throughout development until after birth. A point mutation in the TrkB/PLCγ (TrkbP/P) docking site did not cause loss of geniculate ganglion neurons at any point during development. Animals with a point mutation in both docking sites (TrkbD/D) caused a further decrease in neuron numbers compared to animals with a mutation in only one of the docking sites, similarly to what has previously been shown in Trkb null animals. We concluded that the TrkB/Shc docking site is crucial for determining the survival of geniculate ganglion neurons during mouse gustatory development, while the TrkB/PLCγ docking site does not affect the neuronal survival directly and likely plays a role in maintenance of these neurons. Examining the targeting of geniculate ganglion afferents into the tongue revealed large deficits in innervated neural bud and taste bud numbers in TrkbS/S animals both before and after birth. This was concluded to be reflecting the lack of neuronal survival in this ganglion, a result that was mirrored in TrkbD/D animals. TrkbP/P animals, on the other hand, exhibited a developmental delay in innervation. This was indicated by a low amount of innervated neural buds following the initial innervation period, which was compensated for by a large increase in the number of innervated taste buds by birth. By adulthood, the numbers of taste buds present on the tongues of TrkbP/P animals reached normal numbers compared to control animals. This suggested that the TrkB/PLCγ docking site is involved primarily in innervation. Finally, we examined the morphology of taste buds in newly born and adult animals. We found that the low amount of geniculate ganglion afferents innervating the tongue in TrkbS/S and TrkbD/D animals caused a decrease in size of taste buds. This effect was seen to be partially rescued by adulthood in TrkbS/S animals but not in TrkbD/D animals due to lack of viability. The morphology of taste buds was unaffected in TrkbP/P animals until adulthood, at which point the size of the taste buds was increased. These results are in agreement with previous findings showing dependency of taste bud morphology on the amount of innervation. Overall, our findings show a differential role of TrkB adaptor sites in gustatory development. Despite activated by the same ligands, the docking sites on this receptor are able to exert different influence on signalling pathways downstream of TrkB affecting neuronal survival, targeting and morphology of taste buds.

Midportion Achilles tendinopathy (tendinosis) is a troublesome painful condition, often characterised by pain, local swelling, tenderness and functional disability. Despite extensive research, the pathogenesis is poorly understood and treatment remains challenging. Features related to the peritendinous connective tissue can be of importance. Recently it has been suggested that the plantaris tendon might be involved in this condition. Furthermore, it has been hypothesised that tendon pain and the tendinosis-related tissue changes in tendinopathy might be mediated by signalling substances such as glutamate and acetylcholine. A clinical observation, not scientifically evaluated, has been that unilateral treatment for bilateral Achilles tendinosis can lead to an effect on the contralateral side.      The aim of this work was to examine the morphology and innervation patterns in the plantaris tendon and the peritendinous connective tissue in between the Achillles and plantaris tendons in midportion Achilles tendinopathy, and to evaluate if plantaris tendon removal has an effect on Achilles tendon structure. Another aim was to determine if unilateral treatment for Achilles tendinopathy targeting the peritendinous connective tissue can result in bilateral recovery. Furthermore the presence of non-neuronal cholinergic and glutamate systems was examined.      Sections of plantaris tendons with adjacent peritendinous connective tissue from patients with midportion Achilles tendinopathy were stained for morphology (H&E), and innervation patterns were evaluated using antibodies against general nerve marker (PGP9.5), sensory (CGRP) and sympathetic (TH) nerve fibres and Schwann cells (S-100β). Furthermore immunostainings against non-neuronal aceylcholine (ChAT) and glutamate signalling components (glutamate, VGluT2, NMDAR1) were performed. Plantaris tendon cells were cultured and also stained for glutamate signalling components, and were stimulated with glutamate and glutamate receptor agonist NMDA. Furthermore, Ultrasound Tissue Characterisation (UTC) was used to monitor the integrity of the Achilles tendon collagen structure after plantaris tendon removal.      Plantaris tendons exhibited tendinosis-like tissue patterns such as hypercellularity, collagen disorganisation and large numbers of blood vessels. The peritendinous connective tissue between the plantaris and Achilles tendons contained large numbers of fibroblasts and blood vessels and to some extent macrophages and mast cells. A marked innervation was found in the peritendinous connective tissue and there were also nerve fibres in the loose connective tissue spaces within the tendon tissue proper. Most nerve fibres were identified as sensory fibres. Some nerve fascicles in the peritendinous connective tissue showed absence of axons but homogenous reactions for Schwann cell marker. Tenocytes and cells in the peritendinous connective tissue expressed ChAT, glutamate, VGluT2 and NMDAR1. Tendon cells in vitro expressed VGluT2, NMDAR1 and glutamate. UTC showed significant improvement of Achilles tendon integrity 6 months after surgical plantaris tendon removal and scraping procedure. Eleven out of thirteen patients reported of a bilateral recovery after unilateral surgical treatment.      The results of this work show that plantaris tendons exhibit tendinosis-like tissue changes, internal innervation and features that suggest occurrence of glutamate and acetylcholine production and signalling. Plantaris removal improves Achilles tendon structure suggesting possible compressive/shearing interference between the Achilles and plantaris tendons in tendinopathy. The peritendinous connective tissue shows marked innervation, which thus might transmit pain when being compressed. The partial absence of axons indicates a possible nerve degeneration. On the whole, the study gives new evidence favouring that the plantaris tendon and the peritendinous connective tissue might be of importance for pain and the tendinopathy process in midportion Achilles tendinopathy.

Determination of the physiologic roles of serotonin (5-HT) has long been hampered by the lack of compounds specific for certain of the 5-HT receptor subtypes. The objective of this dissertation was to characterize vascular serotonin receptors in certain arteries and to develop functional assays for the putative 5-HT₁(A) and 5-HT₂ receptors in vascular tissue to test novel compounds. Although 5-HT₁(A) receptor involvement in the 5-HT contraction of the canine basilar artery was previously reported, the 8-OH-DPAT (5-HT₁(A) specific agonist) EC₅₀ values in the canine, rabbit, guinea pig, and bovine basilar arteries studies were not consistent with the presence of 5-HT₁(A) receptors. Studies examining the 5-HT₂ selective antagonist ketanserin, several novel aryltryptamines with a range of affinities, and enantiomers of spiroxatrine, in the 5-HT-contracted rat aorta showed a good correlation between the aorta affinities and the affinities of these compounds at the [³H] ketanserin binding site (defined as 5-HT₂) in the rat frontal cortex. Comparison of the affinities of several known and novel compounds in the rat aorta and the rabbit femoral artery to the [³H] ketanserin site affinities in the frontal cortices of both species showed that the rabbit femoral artery 5-HT₂-like receptor was similar but not identical to either the rat aorta or the CNS sites from either species. The rabbit aorta and the rat femoral artery were then examined to determine if the 5-HT₂ receptor heterogeneity was species or vascular bed specific. The results from all four vascular tissues showed that no two tissues had identical responses to the compounds studied. The rat aorta appeared unique in the lack of agonist activity of RU24969 and the non-competitive antagonism of 5-HT by methysergide, but correlated to the CNS site for the affinities of all compounds. The major finding of the dissertation was the definitive evidence for vascular 5-HT₂ receptor heterogeneity; this subtype was previously thought to be homogeneous. Development of more selective compounds for 5-HT receptor subtypes may lead to greater understanding of the physiological roles of serotonin.

Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.

Rotator cuff injury is a debilitating condition and when injury cannot be managed through non-operative procedures, surgery is required. To determine explanations for the failure of surgery to restore joint function, human muscle biopsies of supraspinatus were studied compared to deltoid. Histology showed atrophy and a tendency toward fibrosis/fatty infiltration in injured supraspinatus. Findings from AChR-subunit western blot and Sema3A localization around satellite cells suggest supraspinatus denervation. Nucleotide incorporation to quantify satellite cell activation in culture showed a significant increase in BrdU+(active) satellite cells in supraspinatus treated with a nitric oxide-donor drug, but not deltoid muscle. The application of principal component analysis to these data extracted components that suggest variables assaying muscle atrophy, satellite cell activity, and fibrosis contribute strongly to the observed variability. The results suggest supraspinatus muscle of the injured rotator cuff is atrophic, denervated, possibly subject to fibro-fatty infiltration, and support the idea that treatment could promote growth in atrophic supraspinatus to improve functional outcomes. / October 2015

A cirurgia da remoção da mama, associada à linfadenectomia axilar, tem sido o tratamento tradicional do câncer de mama. Apesar dos seus benefícios, a cirurgia está associada a algumas seqüelas e complicações, entre elas, as alterações de sensibilidade no trajeto do nervo intercostobraquial. Objetivos: Analisar, através do estesiômetro de Semmes-Weinstein, as alterações de sensibilidade no trajeto do nervo intercostobraquial, em mulheres que realizaram linfadenectomia axilar, por câncer de mama; identificar os descritores de alteração de sensibilidade, segundo a percepção dessas mulheres; comparar a sensibilidade entre mulheres com e sem linfadenectomia axilar; comparar a percepção da sensibilidade de mulheres com linfadenectomia com a resposta obtida com a utilização do estesiômetro. Métodos: Foi aplicado o estesiômetro de Semmes-Weinstein para a avaliação da sensibilidade no trajeto do nervo intercostobraquial e um questionário com dados pessoais e sociodemográficos, acerca do tratamento e sobre a percepção da sensibilidade no membro superior. Participaram desse estudo, 94 mulheres, divididas em dois grupos: um grupo composto por 47 mulheres que realizaram linfadenectomia axilar por câncer de mama e um grupo composto por 47 mulheres não diagnosticadas por câncer de mama e sem qualquer tipo de cirurgia nas axilas. Em cada participante foi realizado o teste com o estesiômetro, por duas vezes consecutivas, e no grupo com cirurgia, também foi aplicado o questionário. Resultados: A prevalência de alteração de sensibilidade, avaliada por meio do estesiômetro de Semmes-Weinstein foi alta, mostrando que 85,1% das mulheres do grupo com linfadenectomia axilar tiveram respostas abaixo dos valores considerados de normalidade, baseado nas respostas do grupo sem linfadenectomia axilar. Foi confirmada a repetibilidade na aplicação do estesiômetro, no grupo com linfadenectomia, através do teste de Kappa (p = 0,8696). O sintoma mais relatado foi adormecimento (70,2%). Conclusões: Na população desse estudo, a queixa de alterações de sensibilidade teve alta prevalência e consideramos o estesiômetro um aparelho de fácil aplicação na prática clínica, baixo custo e confiável. Porém, enfatizamos a importância do treinamento do profissional que irá aplicá-lo e a padronização do teste, de acordo com as orientações do fabricante. / The surgery of the removal of the breast associated to the axillary lymph node dissection has been the traditional treatment for breast cancer. In spite of their benefits, the surgery is associated some sequels and complications, among them the sensibility alterations in the region of the intercostobrachial nerve. Objective: To analyze the sensibility alterations around intercostobrachial nerve in women who were operated by axillary lymph node dissection, for breast cancer through the Semmes- Weinstein monofilament testing; to identify the subject headings of sensibility alteration according to those women\'s perception; to compare the sensibility among women with and without axillary lymph node dissection; and to compare the perception of the women\'s sensibility with axillary lymph node dissection with the answer obtained by the Semmes-Weinstein monofilament testing. Methods: The Semmes-Weinstein monofilament testing was applied for the evaluation of the sensibility in the region of the intercostobrachial nerve and a questionnaire with personal data and sociologically demographic aspects, concerning the treatment and about the perception of the sensibility in the superior member. In that study, 94 women took part in, divided in two groups. A group composed by 47 women that were operated by axillary lymph node dissection for breast cancer and a group composed by 47 women no diagnosed by breast cancer and without any axillary surgery. In each participant the test was accomplished with the monofilament for twice consecutive and in the group with surgery the questionnaire was applied also. Results: The prevalence of sensibility alteration, appraised through the Semmes-Weinstein monofilament testing it was high, 85,1% of the group with axillary lymph node dissection had answers below the considered values of normality, based on the answers of the group without axillary surgery. The repeatability was confirmed in the application of the monofilament in the group with axillary lymph node dissection through the test of Kappa (p=0,8696). The symptom more told was numbness (70,2%). Conclusions: In the population of that study, the complaint of sensibility alterations had high prevalence and we considered the Semmes-Weinstein monofilament testing appliance of easy application in practice clinic, low cost and reliable. However we emphatize the importance of the professional\'s training that will apply it and the standardization of the test in agreement with the manufacturer\'s orientations.

by Phillip Y.P. Jen. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1994. / Includes bibliographical references (leaves 97-110). / Abstract --- p.iii / Acknowledgements --- p.x / Chapter 1. --- Review of literature --- p.1 / Chapter 2. --- Materials and Methods --- p.8 / Chapter 2.1 --- Collection and preparation of tissues --- p.8 / Chapter 2.2 --- Immunofluorescence --- p.9 / Chapter 3. --- Results --- p.15 / Chapter 3.1 --- Urinary bladder --- p.15 / Chapter 3.1.1 --- Bladder detrusor muscle --- p.15 / Chapter 3.1.2 --- Intramural ureters and superficial trigone --- p.17 / Chapter 3.1.3 --- Bladder mucosa --- p.19 / Chapter 3.1.4 --- The bladder neck --- p.20 / Chapter 3.2 --- Vas deferens and seminal vesicle --- p.22 / Chapter 3.2.1 --- The smooth muscle coat --- p.30 / Chapter 3.2.2 --- The mucosa --- p.24 / Chapter 3.3 --- Prostate --- p.26 / Chapter 3.4 --- Urethra --- p.30 / Chapter 3.4.1 --- Rhabdosphincter --- p.31 / Chapter 3.4.2 --- Smooth muscle coat and lamina propria --- p.32 / Chapter 3.5 --- Autonomic ganglia and paraganglia --- p.34 / Chapter 4. --- Discussion --- p.70 / Chapter 4.1 --- Urinary bladder --- p.70 / Chapter 4.2 --- Vas deferens & seminal vesicle --- p.81 / Chapter 4.3 --- Prostate --- p.84 / Chapter 4.4 --- Autonomic ganglia --- p.87 / Chapter 5. --- Suggestions for further study --- p.93 / Chapter 6. --- References --- p.101

Chow Tsun Cheung, Louis. / "May 2000." / Thesis (M.D.)--Chinese University of Hong Kong, 2000. / Includes bibliographical references. / 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. / Abstracts in English and Chinese.

Philip Y.P. Jen. / Thesis (Ph.D.)--Chinese University of Hong Kong, 1996. / Includes bibliographical references (p. 94-111). / 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.

Bibliography: leaves 152-170.