by Sze, Lung Yam. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1994. / Includes bibliographical references (leaves 106-112). / Abstract --- p.i / Acknowledgements --- p.iv / Contents --- p.v / Chapter Chapter 1 --- General Introduction / Chapter 1.1. --- Morphology of the Mammalian Somite and Limb --- p.1 / Chapter 1.1.1. --- The Somite --- p.1 / Chapter 1.1.2. --- The Limb --- p.3 / Chapter 1.2. --- Somite-Limb Relationship --- p.5 / Chapter 1.2.1. --- Somite Contribution to the Appendicular Musculature --- p.5 / Chapter 1.2.2. --- Somite Contribution to Limb Morphogenesis --- p.6 / Chapter 1.3. --- Control of Directionality of Somitic Cell Migration in Appendicular Environment --- p.8 / Chapter 1.4. --- Reasons and Objective of the Present Study --- p.10 / Chapter Chapter 2 --- The Origin of the Mammalian Limb Skeletal Muscles / Chapter 2.1. --- Introduction --- p.16 / Chapter 2.2. --- Materials and Methods --- p.19 / Chapter 2.2.1. --- Embryo Collection --- p.19 / Chapter 2.2.2. --- Isolation of Somites --- p.20 / Chapter 2.2.3. --- DiI-labelling of Rat Donor Somites --- p.21 / Chapter 2.2.4. --- Somite Transplantation --- p.21 / Chapter 2.2.5. --- Embryo Culture --- p.22 / Chapter 2.2.6. --- Analysis of Cultured Embryos --- p.22 / Chapter 2.2.7. --- Cryosection --- p.23 / Chapter 2.2.8. --- Limb Explant Cultures --- p.23 / Chapter 2.2.9. --- Immunohistochemistry --- p.24 / Chapter 2.2.10. --- X-gal Staining --- p.25 / Chapter 2.2.11. --- Histology --- p.25 / Chapter 2.3. --- Results --- p.27 / Chapter 2.3.1. --- Gross Morphology of Cultured Embryos --- p.27 / Chapter 2.3.2. --- Distribution of DiI Labelled Somitic Cells in Rat Embryos --- p.27 / Chapter 2.3.3. --- Histogenetic Potential of Labelled Somitic Cells in the Limbs --- p.29 / Chapter 2.3.4. --- Chimeaic Limb Culture --- p.30 / Chapter 2.4. --- Discussion --- p.32 / Chapter 2.4.1. --- Relationship Between the Somites and the Limb Musculature in Rat Embryos --- p.33 / Chapter 2.4.2. --- Myogenic Potential of Somitic Cells in the Mouse Limb Bud --- p.36 / Chapter 2.4.3. --- The Regulatory Potentials of Mammalian Somites --- p.37 / Chapter Chapter 3 --- The Migration of Somitic Cells into the Mammalian Fore- limb Bud / Chapter 3.1. --- Introduction --- p.52 / Chapter 3.2. --- Materials and Methods --- p.57 / Chapter 3.2.1. --- Embryo Collection --- p.57 / Chapter 3.2.2. --- Embryo Culture and Analysis of Cultured Embryos --- p.58 / Chapter 3.2.3. --- Experimental Series I --- p.58 / Chapter A. --- Micro-injection of DiI --- p.58 / Chapter B. --- Explant Cultures of Rat Fore-limb Bud --- p.59 / Chapter C. --- Histology and Immunohistochemistry --- p.60 / Chapter 3.2.4. --- Experimental Series II --- p.61 / Chapter A. --- Preparation of Conditioned and Unconditioned Medium --- p.61 / Chapter B. --- Coating of Nucleopore Membrane with Fibronectin --- p.62 / Chapter C. --- Preparation of Somitic Cells --- p.62 / Chapter D. --- Analysis of Chemotatic Effect --- p.63 / Chapter 3.2.5. --- Experiment Series III --- p.64 / Chapter A. --- Micro-injection of Latex Beads --- p.64 / Chapter B. --- Isolation of Somatopleure and Transplantation --- p.64 / Chapter C. --- "Somite Isolation, Labelling, and Transplantation" --- p.65 / Chapter D. --- Histology --- p.65 / Chapter 3.3. --- Results --- p.66 / Chapter 3.3.1. --- Development of Embryos In vitro --- p.66 / Chapter 3.3.2. --- Experimental Series I --- p.66 / Chapter A. --- Distribution of Somitic Cells in DiI Injected Embryos --- p.66 / Chapter B. --- Histogenetic Potential of Limb Explants Cultured Under the Kidney Capsule --- p.68 / Chapter C. --- Histogenetic Potential of Limb Explants Cultured In vitro --- p.69 / Chapter 3.3.3. --- Experimental Series II --- p.70 / Chapter A. --- Chemotatic Behaviour of Somitic Cells --- p.70 / Chapter 3.3.4. --- Experimental Series III --- p.71 / Chapter A. --- Ability of Latex Beads to Invade the Limb Bud --- p.71 / Chapter B. --- Distribution Pattern of Somatopleural and Somitic Cells --- p.71 / Chapter 3.4. --- Discussion --- p.75 / Chapter 3.4.1. --- Experimental Series I --- p.75 / Chapter A. --- Distribution of Somitic Cells in DiI-Injected Rat Embryos --- p.75 / Chapter B. --- Histogenetic Potential of Rat Fore-limb Bud --- p.77 / Chapter 3.4.2. --- Experimental Series II --- p.80 / Chapter A. --- Chemotatic Behaviour of Somitic Cells --- p.80 / Chapter 3.4.3. --- Experimental Series III --- p.80 / Chapter A. --- Ability of Latex Beads to Invade the Limb Bud --- p.80 / Chapter B. --- Ability of Somatopleure and Somite to Invade Limb Bud --- p.83 / Chapter 3.4.4. --- Conclusion --- p.83 / References --- p.106 / Appendix --- p.113

by Tang Mei Kuen. / "May 1999." / Thesis (Ph.D.)--Chinese University of Hong Kong, 1999. / Includes bibliographical references (p. 134-179). / 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.

Orientador: Lúcia Elvira Alvares / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-17T20:53:58Z (GMT). No. of bitstreams: 1 Peterlini_DennerJefferson_M.pdf: 3626461 bytes, checksum: 754caba3e8964b3c91965dc6f4912b32 (MD5) Previous issue date: 2011 / Resumo: Os membros de vertebrados representam uma aquisição importante do grupo, os quais possibilitaram a expansão destes pela Biosfera. As bases moleculares do desenvolvimento dos membros estão sob intensa investigação, e o papel de diversos genes e moléculas de sinalização começam a ser bem compreendidos tanto no contexto do estabelecimento de seus eixos quanto da padronização dos tecidos e estruturas. A família dos genes Dapper (Dpr) tem sido associada a diversos processos da embriogênese de vertebrados, desde a coordenação de movimentos morfogenéticos durante a gastrulação à morfogênese de estruturas tão distintas quanto encéfalo, olhos e coração. No entanto, pouco se sabe sobre o papel destes genes no desenvolvimento dos membros, um sítio marcante de sua expressão durante a embriogênese de vertebrados. Resultados preliminares obtidos com emprego de hibridação in situ em embrião de galinha no nosso laboratório já haviam mostrado a expressão destes genes nos membros, e isto sugeriu que eles pudessem desempenhar um papel importante na ontogênese destas estruturas. assim, os padrões de expressão dos genes Dpr1 e Dpr2 entre os estádios HH24 e HH34 da ontogênese de aves foram caracterizados por meio de hibridação in situ neste trabalho. Também foram avaliadas a expressão dos marcadores moleculares MyoD para desenvolvimento de músculo esquelético, e Sox9 para desenvolvimento de cartilagem, bem como foi feita coloração dos membros com alcian blue, que evidência matriz extra-celular de tecido cartilaginoso. Os resultados obtidos revelaram que, no estádio HH24, a expressão de Dpr1 está presente no mesênquima proximal e medial dos membros anterior e posterior, e ausente da região distal (autópode). Neste estádio, a expressão de Dpr2 é claramente associada à agregação das células mesenquimais em condensações pré-condrogênicas. No estádio HH25, transcritos de Dpr1 e Dpr2 foram localizados pela primeira vez no autópode, delimitando uma região com o formato dos moldes cartilaginosos dos dígitos em formação. No estádio HH28, o padrão de expressão de Dpr1 ainda acompanha o contorno dos dígitos, além de serem observados altos níveis de expressão nos precursores dos tarsos e carpos. Por sua vez, Dpr2 é expresso fortemente nos dígitos 1 e 5 dos membros anterior e posterior, bem como nos blastemas dos dígitos posteriores. Finalmente, no estádio HH34, transcritos Dpr1 e Dpr2 estão concentrados nas regiões das articulações dos membros em desenvolvimento, enquanto Dpr2 é expresso também em tendões e em anexos ectodérmicos em formação. Este estudo suporta fortemente a hipótese de que os genes Dpr1 e Dpr2 desempenham um papel no processo de condrogênese que antecede a formação dos ossos dos membros de aves, bem como no desenvolvimento de outras estruturas, como articulações, tendões e anexos cutâneos / Abstract: The acquisition of vertebrates limbs represents an important novelty for this group and allowed the expansion of vertebrates through the Biosphere. The molecular basis of limbs development are under intense investigation, and the role of several genes and signaling molecules begin to be understood both within the context of axis determination as well as in the patterning of tissues and structures. The Dapper (Dpr) gene family has been associated with different processes of vertebrates embryogenesis, from the coordination of morphogenetic movements during gastrulation to morphogenesis of structures as different as brain, eyes and heart. However, nothing is known about the role of these genes in limb development, am important domain of Dpr expression during the embryogenesis of vertebrates. Preliminary results of in situ hybridization in chicken embryo obtained in our laboratory had already shown the expression of these genes in limb, suggesting that they could play an important role in the ontogeny of these structures. Thus, in this study the Dpr1 and Dpr2 expression pattern was characterized by in situ hybridization between stages HH24 and HH34 of chicken development. We also determined the expression of the molecular markers MyoD (skeletal muscle) and Sox9 (cartilage) and stained limbs at the different stages with alcian blue, that labels the extracellular matrix of cartilage. The results revealed that, at stage HH24, Dpr1 expression is observed in the proximal and medial mesenchyme in the fore and hindlimb buds but avoids the autopod. At this stage, the expression of Dpr2 is clearly associated with mesenchymal condensations of pre-chondrogenic cells. At stage HH25, Dpr1 and Dpr2 transcripts were found for the first time in the autopod, delimiting a region with the shape of the cartilaginous templates of the developing digits. At stage HH28, Dpr1 is still expressed around the developing digits, and transcripts are found at high levels in the tarsi and carpi precursors. In turn, Dpr2 is expressed strongly in the first and fifth digits of the forelimbs and hindlimbs, as well as in the digit blastemas. Finally, at stage HH34, Dpr1 and Dpr2 transcripts are concentrated in the developing joints, while Dpr2 is also expressed in ectodermal tendons and developing skin appendages. This study strongly supports the hypothesis that Dpr1 and Dpr2 play a role in the process of chondrogenesis before the formation of the limb bones in birds as well as the development of other structures such as tendons and skin appendages / Mestrado / Histologia / Mestre em Biologia Celular e Estrutural

Orientador: Lúcia Elvira Alvares / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-18T00:32:50Z (GMT). No. of bitstreams: 1 Sensiate_LucimaraAparecida_M.pdf: 8827752 bytes, checksum: 66afd653eb6cf0a2ff14cc79c158cfc1 (MD5) Previous issue date: 2011 / Resumo: Dentre as moléculas envolvidas na sinalização molecular durante o desenvolvimento, têm papel de destaque aquelas envolvidas nas vias de sinalização Wnt e TGF-ß. Trabalhos demonstram que as proteínas Dapper (Dpr) são capazes de modular tais vias de sinalização. A família de genes Dpr é constituída por três componentes: Dpr1, Dpr2 e Dpr3 em peixe-zebra, camundongo e humanos e Dpr1 e Dpr2 em aves. Os genes Dpr têm sido associados a movimentos morfogenéticos durante a gastrulação, especificação do mesoderma, morfogênese do encéfalo, coração e olhos. Apesar da grande relevância desta família gênica para o desenvolvimento, pouco se sabe sobre o padrão de expressão de tais genes em mamíferos e aves. Este projeto de pesquisa teve como objetivo principal a caracterização do padrão de expressão dos genes Dpr ao longo do desenvolvimento embrionário de camundongo e com maiores detalhes, a caracterização do padrão de expressão dos genes Dpr durante a ontogênese dos membros em embriões de camundongo e galinha. O padrão de expressão foi determinado através de ensaios de hibridação in situ whole mount e em cortes de parafina. Para permitir comparações, hibridações in situ foram realizadas com marcadores para o desenvolvimento de cartilagem, tendão e músculo. Como resultado, observamos que os genes Dpr são expressos em seis domínios chave (tubérculo genital, membros, focinho, somitos, hérnia umbilical fisiológica e encéfalo) durante o desenvolvimento de camundongo. A análise do padrão de expressão sugere fortemente o envolvimento dos genes Dpr na ontogênese dos membros em embriões de galinha e camundongo. Contudo, a expressão destes genes é bastante diferente entre estes dois organismos. O gene Dpr1 parece estar envolvido com a formação de elementos da articulação e do pericôndrio durante o desenvolvimento dos membros em embriões de camundongo e galinha. O gene Dpr2 possui expressão bastante difusa durante o desenvolvimento dos membros em embriões de camundongo. Em contrapartida, na galinha, Dpr2 possui expressão localizada no blastema dos dígitos e articulações do autópode. O gene Dpr3 possui expressão difusa durante o desenvolvimento dos membros. Apesar disto, os domínios de expressão identificados sugerem que Dpr3 esteja relacionado com o desenvolvimento dos dígitos e articulações / Abstract: Among the molecules involved in molecular signaling during development, those involved in the Wnt and TGF-ß pathways are particularly important. Studies have shown that the Dapper protein family (Dpr) can modulate Wnt and TGF-ß signaling. The Dpr gene family consists of three members: Dpr1, Dpr2 and Dpr3 in zebrafish, mice and humans, and only two: Dpr1 and Dpr2, in birds. Dpr genes have been associated with morphogenetic movements during gastrulation, mesoderm specification, morphogenesis of the brain, heart and eyes. Despite the great relevance of this gene family during development, little is known about the expression pattern of the Dpr genes in mammals and birds. This research project had as main aims to characterize the expression pattern of the Dpr genes during mouse embryonic development and, in more detail, their expression during limb ontogenesis in mouse and chicken embryos. The expression pattern was determined by in situ hybridization in whole mount and in paraffin sections. To allow comparisons, were performed in situ hybridization with markers for the development of cartilage, tendon and muscle. Our results indicate that Dpr genes are expressed in six key areas (genital tubercle, limbs, nose, somites, brain and physiological umbilical hernia) during development of mouse. The expression pattern in the limbs strongly suggests that the Dpr genes work in limbs development in chicken and mouse embryos. However, the expression pattern of these genes is different in these two organisms. Dpr1 seems to be involved in joint and perichondrium formation during limb development in mouse and chicken embryos. For Dpr2, the expression was diffuse during limb development in mouse embryos. In contrast, chicken Dpr2 has localized expression in the digits and joints. Dpr3 gene has diffuse expression during limb development. However, its expression domains suggest that Dpr3 is related to digits and joints development / Mestrado / Biologia Celular / Mestre em Biologia Celular e Estrutural

Wong, Wan Man. / Thesis submitted in: December 2006. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 85-98). / Abstracts in English and Chinese. / Abstract --- p.i / Abstract in Chinese --- p.iii / Acknowledgements --- p.v / Lists of Figures and Tables --- p.vi / Table of Abbreviations --- p.xi / Table of Contents --- p.xv / Chapter Chapter I --- Introduction / Chapter 1.1 --- Brain and Reproductive Organ Expressed Gene --- p.1 / Chapter 1.2 --- Programmed cell death --- p.4 / Chapter 1.3 --- Limb development in mouse --- p.8 / Chapter 1.4 --- Role of BRE in apoptosis --- p.12 / Chapter 1.5 --- Role of programmed cell death in interdigital tissue regression --- p.14 / Chapter 1.6 --- Aim of study --- p.17 / Chapter Chpater II --- Materials and methods / Chapter 2.1 --- Mice --- p.18 / Chapter 2.2 --- In-situ hybridization / Chapter 2.2.1 --- Histology --- p.18 / Chapter 2.2.2 --- Preparation of riboprobe for in-situ hybridization --- p.19 / Chapter 2.2.3 --- In-situ hybridization --- p.20 / Chapter 2.3 --- Interdigital tissue culture --- p.21 / Chapter 2.4 --- Gene interference / Chapter 2.4.1 --- Construction of Bre-siRNA --- p.22 / Chapter 2.4.2 --- siRNA transfection of cultured interdigital cells --- p.23 / Chapter 2.5 --- Semi-quantitative RT-PCR / Chapter 2.5.1 --- Sample collection of interdigital cells and explants --- p.23 / Chapter 2.5.2 --- RNA isolation and extraction --- p.24 / Chapter 2.5.3 --- Reverse-transcription and cDNA synthesis --- p.25 / Chapter 2.5.4 --- Polymerase chain reaction --- p.26 / Chapter 2.6 --- Assay of cell viability by MTT --- p.28 / Chapter 2.7 --- Comparative proteomics --- p.30 / Chapter 2.7.1 --- Collection of interdigital cells --- p.30 / Chapter 2.7.2 --- Preparation of cell lysate --- p.31 / Chapter 2.7.3 --- Assay of protein concentration in cell lysate --- p.31 / Chapter 2.7.4 --- Two-dimensional gel electrophoresis --- p.33 / Chapter 2.7.5 --- Protein identification by mass fingerprinting --- p.36 / Chapter 2.8 --- Statistical Method --- p.38 / Chapter Chapter III --- Results / Chapter 3.1 --- Spatial and temporal expression of Bre in murine embryonic hindlimbs --- p.39 / Chapter 3.2 --- Expression of Bre isoforms in interdigital tissues --- p.45 / Chapter 3.3 --- Silencing of Bre expression by siRNA in interdigital cells --- p.49 / Chapter 3.4 --- Effect on viability of Bre-silenced interdigital cells by siRNA --- p.51 / Chapter 3.5 --- Comparative proteomic profile of Bre-silenced interdigital cultured cells --- p.53 / Chapter 3.6 --- Identification of proteins that were differentially expressed by MALDI- TOF --- p.71 / Chapter 3.7 --- The mRNA levels of proteins identified that were differentially expressed --- p.74 / Chapter Chapter IV --- Discussion --- p.77 / References --- p.85 / Appendices --- p.99 / Publication --- p.108