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RNA interference (RNAi) is a conserved mechanism in a wide range of eukaryotes. Introduction of synthetic dsRNA could specifically target suppression of a gene or could result in off-target silencing of another gene due to sequence similarity. To verify if the observed phenotype in an RNAi transgenic line is due to silencing of a specific gene or if it is due to another nontarget gene, a synthetic gene complementation approach could be used. Synthetic gene complementation described in this method uses the technology of synthesizing a variant of a native gene (used in RNAi silencing) to maximize the difference in DNA sequences while coding for the exact same amino acids as the original native gene. This is achieved through the use of alternate codons. The new variant gene is expressed in the original RNAi transgenic lines and analyzed for complementation of the RNAi phenotype. Complementation of the RNAi-induced phenotype will indicate gene-specific silencing and not off-target silencing.

A prematuridade se caracteriza por ser qualitativamente distinta do nascimento de termo, em função das intercorrências neonatais, do impacto do período de permanência na incubadora e da influência da ação da gravidade no desenvolvimento postural, do equilíbrio e da locomoção. Este estudo teve como OBJETIVOS: 1- comparar a coordenação motora axial de lactentes pré-termo (RNPT) saudáveis, com a de lactentes de termo (RNT), por meio dos escores da Alberta Infant Motor Scale (AIMS), segundo a idade cronológica (ICr) e corrigida (ICo), a partir do termo até a utilização da marcha independente como principal meio de locomoção; 2- verificar período de aquisição, entre RNPT e RNT, segundo ICo, de 7 itens avaliados pela AIMS (transição de quatro apoios para sentado; engatinhar; segurar o(s) pé(s) em supino; sentar-se independente; transição de sentado para quatro apoios; ficar em pé sem apoio; marcha independente); 3- determinar a influência de variáveis biológicas e sóciodemográficas para obtenção da marcha em RNPT. MÉTODOS: estudo coorte, prospectivo, observacional, mensal e comparativo entre 101 RNPT (grupo de estudo) e 52 RNT (grupo controle). Foram ajustados modelo de regressão beta e equações de estimação generalizada para curvas de ICr e ICo, a fim de comparar a média mensal dos escores entre os grupos. Para comparar as idades de aquisição dos sete itens da AIMS entre os RNT e RNPT, foram utilizadas estimativas de Turnbull da distribuição da idade e modelo de taxa de falhas proporcionais de Cox para censuras intervalares. Para análise estatística dos resultados das médias mensais entre os grupos e as idades de aquisição dos itens, recorreu-se à construção de intervalo de confiança (IC). Para análise das variáveis prognósticas no tempo da aquisição da marcha nos RNPT, análise de sobrevivência para censuras intervalares e modelo de regressão Weibull foram utilizados. O nível de significância considerado foi < 5%. RESULTADOS: Finalizaram o estudo 77 RNPT e 49 RNT. Os escores segundo ICr dos RNPT demonstraram ser estatisticamente inferiores em relação aos dos RNT. Segundo a ICo, os escores dos RNPT tornaram-se equivalentes aos dos RNT, não demonstrando diferença estatística significativa. Dos 7 itens analisados, somente em pé sem apoio e a marcha foram adquiridos mais tardiamente pelos RNPT. As variáveis biológicas com influência significante no tempo de aquisição da marcha foram: peso ao nascimento (PN), estatura ao nascimento (EN) e permanência no hospital após nascimento. CONCLUSÃO: A coordenação motora axial dos RNPT deve ser avaliada segundo ICo para não ser subestimada. A aquisição da marcha e a posição em pé sem apoio se desenvolvem mais tardiamente nos RNPT. Além disto, a aquisição da marcha em RNPT foi tão mais tardia quanto menor o PN e EN, assim como o longo período de internação após o nascimento retardaram a marcha nos RNPT. / The prematurity characterizes to be qualitatively different from term delivery, due to neonatal morbidities, impact of the long period in the incubator and the influence of gravidity in the postural development. This study has as OBJECTIVES:1- to compare the gross motor development in healthy preterm infants (PT) with term infants (T), using Alberta Infant Motor Scale (AIMS) scores, according to corrected and chronological ages, from term to walking independently; 2- To compare the age attainment of seven AIMS items, according to corrected age (four-point kneeling to sitting; reciprocal creeping; hands to feet in supine; sitting independently; sitting to four-point kneeling; stand alone; early stepping) between PT and T infants; 3- To study biologic and sociodemographic factors that affect walking attainment in PT. METHODS: cohort, prospective, observational monthly and comparative study between 101 PT and 52 T. In order to compare descriptively mean monthly scores, beta regression models and general estimated equations were used to adjust the chronological and corrected age graphics and for statistics purposes, the confidence interval of monthly mean scores were used. Turnbull estimation of age distribution and Cox´s proportional hazards model were used to compare the age in each seven items between groups. For prognostic factors of age walking attainment in PT, methods of Kaplan-Meyer and Weibull regression model were used. The level of significance was considered significant, if p < 5%. RESULTS: 77 PT and 49 T infants finished the study. Chronological age scores of PT were statistically lower in comparison with T scores. According to corrected age, PT monthly mean scores were not statistically different from T scores. Of seven analysed items, only standing alone and early stepping were attained later in PT infants. The variables that seemed to influence statistically in age of walking attainment were: birth weight and stature and duration of hospitalization. CONCLUSION: Gross motor development of PT infants should be assessed according to corrected age, in order not to be underestimated. The both milestones standing alone and early stepping developed later in PT infants. The lower the birth stature and weight were, the longer it took for the PT infants to attain walking, while the longer the PT newborns stayed hospitalized, the later they started walking.

The poultry-to-human transmission of the influenza virus and the recent H1Nl influenza pandemic have become major concerns worldwide. The nucleoprotein (NP) of influenza virus binds the RNA genome and plays essential role in transcription and replication during the virus life cycle. / The study leads to a better understanding towards the RNP organization of influenza virus and provides information for the future design of anti-influenza agents. / We have also shown, by RNP reconstitution assay and co-immunoprecipitation, that the interaction between NP and PB2 is crucial for the proper functioning of the RNP. The functional association of NP and PB2 requires either the PB2 host-determining residue lysine-627 or arginine-630 with the latter involving NP arginine-150 also. Using SPR, we have demonstrated that both residues take part in the direct protein-protein interaction, without the involvement of RNA. These results suggest a dual interaction mechanism between NP and PB2. This may confer replication advantages to the virus, as either one can give an active RNP and explains the increased virulence of avian influenza viruses carrying the E627K mutation in mammalian cells. In addition, our findings identify the NP-PB2 interacting surface, with the PB2 627/630 region facing the RNA binding groove of NP. / We have determined the 3.3 A crystal structure of H5N1 NP, which is composed of head and body domains and a tail loop. Using surface plasmon resonance (SPR), we found the basic loop (residues 73-91) and arginine-rich groove, but mostly a protruding element centering at R174 and R175, to be important in RNA binding. Ribonucleoprotein (RNP) reconstitution assay with these multiple-point and deletion mutants indicate their functional importance towards the transcription-replication activities of the virus polymerase. Single-point mutations at these concerned regions do not have a significant effect on their RNP activities, suggesting that NP mediates RNA-binding through multiple residues. / Ng, Ka Leung. / Adviser: Pang Chui Shaw. / Source: Dissertation Abstracts International, Volume: 73-06, Section: B, page: . / Thesis (Ph.D.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 121-136). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [201-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

by Lau Hoi Lun. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1998. / Includes bibliographical references (leaves 83-96). / Abstract also in Chinese. / Abstract --- p.i / Acknowledgements --- p.iv / Abbreviations --- p.v / Table of contents --- p.vi / Chapter Chapter 1 --- Introduction / Chapter 1.1 --- Endocrine control of normal and abnormal growth of prostate --- p.1 / Chapter 1.1.1 --- Androgen regulation of prostate gland --- p.1 / Chapter 1.1.2 --- Estrogen regulation of prostate gland --- p.4 / Chapter 1.2 --- Gonadotropin-releasing hormone plays a central role in reproduction --- p.6 / Chapter 1.2.1 --- GnRH gene --- p.7 / Chapter 1.2.2 --- GnRH receptor --- p.9 / Chapter 1.3 --- Therapeutic strategies using GnRH analogs to treat prostate cancer --- p.12 / Chapter 1.4 --- Expression of GnRH or its receptor in reproductive tissues --- p.12 / Chapter 1.4.1 --- Expression of GnRH in reproductive --- p.13 / Chapter 1.4.2 --- Expression of GnRH and its receptor in pituitary and reproductive tissues --- p.13 / Chapter 1.5 --- Animal models for the study of prostate cancer --- p.15 / Chapter 1.5.1 --- Nobel rat inducible model --- p.15 / Chapter 1.5.2 --- Androgen dependent rat Dunning prostatic tumor --- p.16 / Chapter 1.5.3 --- Androgen-independent prostatic carcinoma line of Noble rat --- p.18 / Chapter 1.6 --- Aim of study --- p.18 / Chapter Chapter 2 --- Materials and Methods / Chapter 2.1 --- Origin and supply of Nobel rat --- p.20 / Chapter 2.2 --- Induction of dysplasia in Nobel rat prostate gland by long-term treatment with steroids --- p.20 / Chapter 2.2.1 --- Chemicals --- p.20 / Chapter 2.2.2 --- Preparation of steroid hormone-filled Silastic tubings --- p.20 / Chapter 2.2.3 --- Surgical implantation of Silastic® tubings --- p.21 / Chapter 2.2.4 --- Protocols of hormonal treatments --- p.21 / Chapter 2.3 --- Androgen- dependent Dunning rat prostatic adenocarcinoma --- p.22 / Chapter 2.4 --- Androgen- independent prostatic carcinoma line (ALT) of Noble rat --- p.22 / Chapter 2.5 --- Detection of mRNA expression of gonadotropin- releasing hormone (GnRH) in normal and neoplastic rat prostates --- p.23 / Chapter 2.5.1 --- Preparation of tissue for total RNA extraction --- p.23 / Chapter 2.5.2 --- Total RNA extraction --- p.24 / Chapter 2.5.3 --- Reverse-transcription Polymerase Chain Reaction (RT-PCR) --- p.25 / Chapter 2.5.4 --- Purification of DNA fragments from agarose gels --- p.27 / Chapter 2.5.5 --- Subcloning of DNA into vector --- p.27 / Chapter 2.5.6 --- Nucleotide sequencing --- p.30 / Chapter 2.5.7 --- Southern blot analysis --- p.32 / Chapter 2.5.7.1 --- Southern blotting --- p.32 / Chapter 2.5.7.2 --- Preparation of α-32P-dCTP labelled GnRH probe --- p.32 / Chapter 2.5.7.3 --- Hybridization --- p.33 / Chapter 2.6 --- Detection of mRNA expression of gonadotropin-releasing hormone receptor (GnRH-R) in normal and neoplastic rat prostates --- p.34 / Chapter 2.6.1 --- Cloning of GnRH-R cDNA and synthesis of its probe --- p.34 / Chapter 2.6.2 --- Detection of GnRH receptor mRNA expression in normal and dysplastic Nobel rat prostates by Southern blot --- p.36 / Chapter 2.6.3 --- Detection of GnRH receptor mRNA expression in Dunning tumor --- p.37 / Chapter 2.6.4 --- Detection of the GnRH receptor mRNA expression in AIT tumor by RT-PCR --- p.37 / Chapter Chapter 3 --- Results / Chapter 3.1 --- Detection of mRNA expression of gonadotropin-releasing hormone (GnRH) in normal and neoplastic rat prostates --- p.38 / Chapter 3.1.1 --- Reverse -transcription Polymerase Chain Reaction (RT-PCR) --- p.38 / Chapter 3.1.2 --- Purification of DNA fragments amplified by PCR from the agarose gel --- p.38 / Chapter 3.1.3 --- Subcloning of DNA into vector --- p.39 / Chapter 3.1.4 --- Nucleotide sequencing --- p.39 / Chapter 3.1.5 --- Southern-blot analysis --- p.39 / Chapter 3.2 --- Detection of gonadotropin-releasing hormone receptor mRNA expression in normal and neoplastic rat prostates --- p.40 / Chapter 3.2.1 --- Cloning of gonadotropin-releasing hormone receptor (GnRH) cDNA and synthesis of probe from the normal Noble rat pituitary gland --- p.40 / Chapter 3.2.2 --- Detection of GnRH receptor mRNA expression in normal and dysplastic Nobel rat prostates --- p.42 / Chapter 3.2.3 --- Detection of GnRH receptor mRNA expression in rat Dunning tumor by PCR --- p.43 / Chapter 3.2.4 --- Detection of GnRH receptor mRNA expression in AIT tumor --- p.43 / Chapter Chapter 4 --- Discussion / Chapter 4.1 --- Detection of mRNA expression of gonadotropin-releasing releasing hormone(GnRH) in normal and neoplastic rat prostates --- p.69 / Chapter 4.1.1 --- Expression of GnRH mRNA in normal Nobel rat prostate gland --- p.69 / Chapter 4.1.2 --- Expression of GnRH mRNA in dysplastic Nobel rat prostate --- p.71 / Chapter 4.1.3 --- Expression of GnRH mRNA in androgen-dependent rat Dunning prostatic tumor --- p.72 / Chapter 4.1.4 --- Expression of GnRH mRNA in AIT tumor --- p.74 / Chapter 4.2 --- Detection of GnRH receptor in normal and dysplastic rat prostates --- p.75 / Chapter 4.2.1 --- Negative expression of GnRH receptor in normal and dysplastic Nobel in rat prostates --- p.75 / Chapter 4.2.2 --- Positive expression of GnRH receptor mRNA in rat Dunning tumor --- p.77 / Chapter 4.2.3 --- Negative expression of GnRH receptor mRNA in ALT tumor --- p.78 / Chapter Chapter 5 --- Summary and Conclusions --- p.80 / References --- p.83

Su Huan Xing. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 79-89). / Abstracts in English and Chinese. / Abstract --- p.i / 中文摘要 --- p.iii / Acknowledgements --- p.iv / Abbreviations Frequently Used --- p.v / Table of contents --- p.vi / Chapter Chapter1 --- General Introduction --- p.1 / Chapter Chapter2 --- Effects of an intravitreal optic nerve graft on the sprouting and regeneration of axotomized retinal ganglion cells --- p.17 / Chapter Chapter3 --- Effects of an intravitreal pre-injured optic nerve graft on the sprouting and regeneration of axotomized retinal ganglion cells --- p.44 / Chapter Chapter4 --- Effects of co-transplantation of an optic nerve graft and a peripheral nerve graft into the vitreous body on the sprouting and regeneration of axotomized retinal ganglion cells --- p.60 / Chapter Chapter5 --- General discussion --- p.74 / References --- p.79 / Tables --- p.90

Wang, Chengdong. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2013. / Includes bibliographical references (leaves 113-134). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

Law Sui-Lin. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 99-109). / Abstracts in English and Chinese. / CONTENTS --- p.i / ABSTRACT --- p.v / 撮要 --- p.vii / ACKNOWLEDGEMENTS --- p.ix / LIST OF FIGURES & TABLES --- p.xi / ABBREVIATIONS --- p.xv / Chapter Chapter 1 --- INTRODUCTION --- p.1 / Chapter 1.1 --- The Transition of Hyperplastic to Hypertrophic Growth During Heart Development --- p.1 / Chapter 1.2 --- The Controversial Capability of Heart Regeneration --- p.3 / Chapter 1.3 --- Challenges in Treating Heart Diseases --- p.5 / Chapter 1.4 --- A New Insight Behind Traditional Chinese Medicine (TCM) for Treating Heart Diseases --- p.7 / Chapter 1.5 --- The Potential Mitogenic TCMs on Cardiomyocytes --- p.10 / Chapter 1.5.1 --- Radix Astragali --- p.11 / Chapter 1.5.2 --- Radix Ophiopogonis --- p.12 / Chapter Chapter 2 --- MATERIALS & METHODS --- p.14 / Chapter 2.1 --- Materials --- p.14 / Chapter 2.2 --- Cell Culture --- p.16 / Chapter 2.2.1 --- Primary neonatal rat cardiomyocytes cell culture --- p.16 / Chapter 2.2.1.1 --- Mayer's hemalum-eosin staining --- p.17 / Chapter 2.2.2 --- Primary rat fibroblasts cell culture --- p.18 / Chapter 2.2.3 --- H9C2 cardiac cell culture --- p.18 / Chapter 2.3 --- TCMs Preparation and Treatment --- p.19 / Chapter 2.3.1 --- Preparation of TCMs powder from aqueous extracts --- p.19 / Chapter 2.3.2 --- Preparation of culture medium with TCMs powder --- p.19 / Chapter 2.3.3 --- Pre-treatment of undifferentiated and differentiated H9C2 cardiac cells with TCMs --- p.20 / Chapter 2.3.4 --- Post-treatment of differentiated H9C2 cardiac cells with TCMs --- p.20 / Chapter 2.4 --- Assessment of DNA Synthesis and Proliferation --- p.21 / Chapter 2.4.1 --- Tritiated thymidine incorporation assay --- p.21 / Chapter 2.4.2 --- 5-Bromo-2'-deoxy-uridine (BrdU) assay --- p.22 / Chapter 2.4.3 --- Cell counting --- p.23 / Chapter 2.4.4 --- Statistical analysis --- p.23 / Chapter 2.5 --- Screening of Differentially Expressed Genes in H9C2 Cells after TCM Treatment by cDNA Microarray --- p.25 / Chapter 2.5.1 --- Total RNA extraction --- p.25 / Chapter 2.5.2 --- RNA labeling --- p.26 / Chapter 2.5.2.1 --- Synthesis of fluorescence labeled probe --- p.26 / Chapter 2.5.2.2 --- Purification of fluorescence labeled probe --- p.27 / Chapter 2.5.3 --- Microarray hybridization --- p.28 / Chapter 2.5.3.1 --- Concentration of fluorescence labeled probe --- p.28 / Chapter 2.5.3.2 --- Hybridization --- p.28 / Chapter 2.5.3.3 --- Post-hybridization treatment --- p.29 / Chapter 2.5.4 --- Data collection --- p.29 / Chapter 2.5.4.1 --- Scanning of slide --- p.29 / Chapter 2.5.4.2 --- Image processing: spots finding and quantification --- p.30 / Chapter 2.5.5 --- Data normalization and analysis --- p.30 / Chapter 2.6 --- Confirmation of Differentially Expressed Genes in H9C2 Cells after TCM Treatment by RT-PCR --- p.32 / Chapter 2.6.1 --- DNase I digestion of total RNA sample --- p.32 / Chapter 2.6.2 --- First-strand cDNA synthesis --- p.32 / Chapter 2.6.3 --- RT-PCR of the candidate genes --- p.33 / Chapter Chapter 3 --- RESULTS --- p.36 / Chapter 3.1 --- Neonatal Primary Rat Cardiomyocytes --- p.36 / Chapter 3.1.1 --- Preparation of high-purity neonatal primary rat cardiomyocytes --- p.36 / Chapter 3.1.2 --- Neonatal primary rat cardiomyocytes ceased to undergo DNA replication after 6-day in vitro culturing --- p.38 / Chapter 3.1.3 --- Both MD and HQ promoted the growth of day 1 primary rat cardiomyocytes in dose- and time-dependent manners --- p.40 / Chapter 3.1.4 --- HQ is more potent than MD in promoting the growth of day 7 primary rat cardiomyocytes --- p.43 / Chapter 3.2 --- H9C2 Cardiac cells --- p.45 / Chapter 3.2.1 --- Proliferative effect of MD and HQ on undifferentiated H9C2 cardiac cells --- p.45 / Chapter 3.2.2 --- Pre-treatment of HQ on H9C2 cardiac cells during differentiation --- p.50 / Chapter 3.2.3 --- Pre-treatment of MD and HQ on differentiated H9C2 cardiac cells --- p.52 / Chapter 3.2.4 --- Post-treatment of MD on differentiated H9C2 cardiac cells…… --- p.55 / Chapter 3.3 --- Primary Rat Fibroblasts --- p.57 / Chapter 3.3.1 --- Proliferative effect of MD and HQ on primary rat fibroblasts --- p.58 / Chapter 3.4 --- Screening of Differentially Expressed Genes in H9C2 Cells after HQ Treatment by cDNA Microarray --- p.60 / Chapter 3.4.1 --- Differentially expressed genes in undifferentiated H9C2 cardiac cells after HQ treatment --- p.60 / Chapter 3.4.2 --- Differentially expressed genes in differentiated H9C2 cardiac cells after HQ treatment --- p.66 / Chapter 3.4.3 --- Comparison of differentially expressed genes in both undifferentiated and differentiated H9C2 cardiac cells after HQ treatment --- p.72 / Chapter 3.5 --- Confirmation of Differentially Expressed Genes in H9C2 Cells after HQ Treatment by RT-PCR --- p.73 / Chapter 3.5.1 --- "Preferential up-regulation of N-G, N-G-dimethylarginine dimethylaminohydrolase mRNA expression level in undifferentiated H9C2 cardiac cells after HQ treatment " --- p.74 / Chapter 3.5.2 --- Preferential up-regulation of heme oxygenase-3 mRNA expression level in undifferentiated H9C2 cardiac cells after HQ treatment --- p.75 / Chapter 3.5.3 --- Preferential up-regulation of cyclin B mRNA expression level in differentiated H9C2 cardiac cells after HQ treatment --- p.76 / Chapter Chapter 4 --- DISCUSSION --- p.77 / Chapter 4.1 --- HQ Being a More Effective Mitogenic TCM than MD on Cardiomyocytes Exerted its Effect in Dose- and Time Dependent --- p.79 / Chapter 4.2 --- Mitogenic Effect of Both MD and HQ might Possibly Due to the Regulation of Intrinsic Factors --- p.82 / Chapter 4.3 --- HQ Rather Than MD Showed a Higher Specificity in Promoting DNA Synthesis in Cardiomyocytes --- p.83 / Chapter 4.4 --- The Differentially Expressed Genes were Supported by The Clinical Functions of HQ --- p.85 / Chapter 4.5 --- Relating the Differentially Expressed Genes with Cardiac Growth and Development --- p.87 / Chapter 4.6 --- The Hypothetic Mechanisms of Action that HQ Exerted on Cardiac Growth and Development --- p.92 / Chapter 4.7 --- Future Prospect --- p.94 / Chapter 4.7.1 --- In vivo study of HQ on the proliferation of rat cardiomyocytes from neonatal to postnatal development --- p.94 / Chapter 4.7.2 --- The study of transgenic mice carrying the target gene regulated by HQ on cardiac growth and development --- p.96 / Chapter 4.7.3 --- The determination of active component of HQ on cardiac growth and development --- p.97 / REFERENCES --- p.99 / APPENDIX --- p.110

Dong Ming. / Thesis submitted in: December 2005. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (leaves 118-152). / Abstracts in English and Chinese. / Abstract (English) --- p.i / Abstract (Chinese) --- p.iii / Acknowledgement --- p.iv / Table of Contents --- p.v / Abbreviation list --- p.xi / Chapter Chapter 1 --- General introduction / Chapter 1.1 --- Preamble --- p.1 / Chapter 1.2 --- Neural Crest Cells (NCCs) --- p.2 / Chapter 1.3 --- Enteric Nervous System (ENS) and Vagal Neural Crest Cells (Vagal NCCs) --- p.4 / Chapter 1.4 --- Sacral Neural Crest Cells (Sacral NCCs) --- p.7 / Chapter 1.5 --- Signalling Mechanisms of Sacral Neural Crest Cells --- p.17 / Chapter 1.6 --- Hirschsprung's Disease (HSCR) --- p.20 / Chapter 1.7 --- Objective of the Study and Contents of the Following Chapters --- p.21 / Chapter Chapter 2 --- Migration from the dorsal neural tube to the pelvic mesenchyme / Chapter 2.1 --- Introduction --- p.24 / Chapter 2.2 --- Materials and Methods --- p.32 / Chapter 2.2.1 --- Animal --- p.32 / Chapter 2.2.2 --- Isolation of embryos from pregnant mice at E9.5 to --- p.32 / Chapter 2.2.3 --- Histological preparation of the caudal segments --- p.33 / Chapter 2.2.4 --- p75 immunohistochemical staining --- p.33 / Chapter 2.2.5 --- Preparation of rat serum --- p.34 / Chapter 2.2.6 --- Preparation of the culture medium --- p.34 / Chapter 2.2.7 --- Preparation of wheat germ agglutinin-gold conjugates (WGA-Au) --- p.35 / Chapter 2.2.8 --- Preparation of CMFDA --- p.35 / Chapter 2.2.9 --- Preparation of DiI --- p.36 / Chapter 2.2.10 --- "Microinjection of WGA-Au, DiI and CMFDA" --- p.36 / Chapter 2.2.11 --- Whole embryo culture --- p.37 / Chapter 2.2.12 --- Examination of cultured embryos --- p.37 / Chapter 2.2.13 --- Histological preparation of WGA-Au labelled embryos --- p.38 / Chapter 2.2.14 --- Silver enhancement staining of the sections of WGA-Au labelled embryos --- p.39 / Chapter 2.2.15 --- Cryosectioning of the embryos labelled with DiI --- p.39 / Chapter 2.2.16 --- p75 immunohistochemical staining of DiI-labelled cells --- p.40 / Chapter 2.3 --- Results --- p.41 / Chapter 2.3.1 --- Observations on embryos developed in vivo --- p.41 / Chapter 2.3.2 --- Closed yolk sac culture vs open yolk sac culture --- p.42 / Chapter 2.3.3 --- Neural crest cell labelling in the caudal part of embryos --- p.43 / Chapter 2.3.4 --- Neural crest cell labelling with DiI in the caudal part of the neural tube followed by in vitro culture from E9.5 to E11.0 --- p.45 / Chapter 2.3.5 --- Neural crest labelling with DiI in the caudal part of the neural tube followed by in vitro culture from E10.5 to E11.5 --- p.46 / Chapter 2.3.6 --- Focal labelling at the levels of the 26th and 29th somites followed by in vitro culture --- p.48 / Chapter 2.3.7 --- p75 immunohistochemical staining on the caudal part of the embryo at E10.5 --- p.49 / Chapter 2.3.8 --- p75 immunohistochemical staining on embryos labelled with DiI --- p.50 / Chapter 2.4 --- Discussion --- p.51 / Chapter 2.4.1 --- Embryos at E9.5 cultured with an intact yolk sac membrane grew better than those with the yolk sac membrane cut open --- p.52 / Chapter 2.4.2 --- Migration at the levels of the 24th to 28th somite --- p.53 / Chapter 2.4.3 --- Migration at the levels of the 29th to 33th somite --- p.58 / Chapter 2.4.4 --- Sacral NCCs migrate along a straight dorsolateral pathway --- p.60 / Chapter 2.4.5 --- "Most of the DiI positive cells are p75 positive, but not all of the p75 positive cells are DiI positive" --- p.62 / Chapter Chapter 3 --- Migration from the pelvic mesenchyme to the hindgut / Chapter 3.1 --- Introduction --- p.65 / Chapter 3.2 --- Materials and Methods --- p.73 / Chapter 3.2.1 --- Isolation of hindguts with or without adjacent tissues from embryos at E10.5 to E13.5 --- p.73 / Chapter 3.2.2 --- Microinjection of DiI into the pelvic mesenchymal tissue of the h indguts --- p.74 / Chapter 3.2.3 --- Preparation of the culture medium --- p.74 / Chapter 3.2.4 --- Preparation of the culture dish --- p.74 / Chapter 3.2.5 --- Gut culture --- p.75 / Chapter 3.2.6 --- Cryosections of the hindguts after in vitro culture --- p.75 / Chapter 3.3 --- Results --- p.76 / Chapter 3.3.1 --- "Hindguts isolated from embryos at E10.5, E11.5, E12.5 and E14.5" --- p.76 / Chapter 3.3.2 --- p75 immunohistochemical staining of the serial sections through the caudal part of the embryos --- p.78 / Chapter 3.3.3 --- Observations on hindgut without pelvic plexus cultured from E11.5 to E14.5 --- p.81 / Chapter 3.3.4 --- "Culture of hindguts with pelvic mesenchyme cultured from E11.5 to E14.25, E14.5 and E15.5" --- p.82 / Chapter 3.3.5 --- Culture of the whole length of the gut tube without pelvic mesenchyme from E11.5 to E15.5 --- p.84 / Chapter 3.3.6 --- Culture of the whole length of the gut tube with pelvic mesenchyme from E11.5 to E15.5 --- p.84 / Chapter 3.3.7 --- "Culture of hindguts with Dil labelling in the pelvic mesenchyme from E11.5 to E14.0, E14.2 5, E14.5 and" --- p.85 / Chapter 3.3.8 --- Culture of the whole length of the gut tube with DiI labelling in the pelvic mesenchyme from E11.5 to E14.5 --- p.86 / Chapter 3.4 --- Discussion --- p.88 / Chapter 3.4.1 --- Development of the hindgut and the urogenital system from E10.5 to E14.5 --- p.88 / Chapter 3.4.2 --- No p75 positive cells were found in the hindgut before E13.5 --- p.89 / Chapter 3.4.3 --- The sacral neural crest cells migrate into the hindgut at around E14.5 --- p.91 / Chapter 3.4.4 --- "The sacral neural crest cells migrated in the serosa, and entered the myenteric plexus prior to populating the submucosal plexus" --- p.95 / Chapter 3.4.5 --- Most of DiI labelled sacral neural crest cells in the hindgut also expressed p75 --- p.98 / Chapter Chapter 4 --- Migration from the neural tube to the hindgut / Chapter 4.1 --- Introduction --- p.101 / Chapter 4.2 --- Materials and Methods --- p.104 / Chapter 4.3 --- Results --- p.106 / Chapter 4.3.1 --- Morphology observations on the hindguts isolated from DiI-labelled embryos --- p.106 / Chapter 4.3.2 --- Distribution of the DiI-labelled cells and p75 positive cells before the culture of the hindgut explant --- p.106 / Chapter 4.3.3 --- Culture of hindgut explanted from Dil-labelled embyos for 3.5 days from E11.0 to E14.5 --- p.107 / Chapter 4.4 --- Discussion --- p.109 / Chapter Chapter 5 --- General discussion and conclusions --- p.113 / References --- p.118 / Figures and Legends --- p.153 / Tables and Graphs --- p.203 / Appendix --- p.209