Call number: LD2668 .T4 1979 G34 / Master of Science

Call number: LD2668 .T4 1979 R617 / Master of Science

Call number: LD2668 .T4 1984 S326 / Master of Science

<p>The phylum Priapulida is a small group of marine worms that is allied with the nematodes, kinorhynchs, loriciferans and nematomorphs in a clade called the Cycloneuralia or Introverta. Together with the arthropods they are generally considered to comprise the Ecdysozoa, a clade of moulting animals. A number of recent priapulid species possess features that resemble the predicted Ecdysozoan ancestor. In addition, recent molecular studies have also shown that they are basal within the Ecdysozoa/Cycloneuralia (Garey 2001, Webster et al. 2006). Their putative basal position thus makes priapulids highly interesting research objects for understanding the evolution of Ecdysozoa. </p><p>Earlier investigations of the early embryology of the priapulid <i>Priapulus caudatus</i> are critically revised with the aid of modern techniques and equipment, confirming earlier studies that the early cleavages are highly symmetrical, total, subequal, radial and stereotypical. New results show that up to the sixth cleavage, the spindles are oriented along the animal/vegetal axis at both poles. This unique cleavage pattern has only limited similarities to other animals. During the sixth cleavage two cells move inwards and gastrulation commences. If the mesoderm is derived from both cells, its origin differs from that of many other protostomes.</p><p>Two previously undescribed larval stages of <i>P. caudatus</i>; the light bulb shaped hatchling and the first lorica larva are described. The second lorica larva superficially resembles the previously described type 2 lorica larva (Higgins et al 1993). Differences between the second lorica larva and the type 2 lorica larva, with respect to possible ecophenotypical variation and sub-specialization, are described. </p><p>Preliminary data are presented on musculature development of <i>P. caudatus</i>. Preliminary data have also been obtained on the early development of a second priapulid, <i>Halicryptus spinulosus</i>. Comparison of <i>Halicryptus</i> and <i>Priapulus</i> may help to resolve developmental ground pattern of the priapulids.</p>

O Tubarão azul (Prionace glauca), popularmente conhecido como cação-azul dentre todas as espécies de tubarão é a mais abundante no ambiente marinho, podendo ser encontrado em todos os mares. Com a diversidade das espécies, a descrição de qualquer fígado especifico, dificilmente poderá ser utilizada como um modelo. Juntamente com essa variabilidade, algumas características fisiológicas dos peixes contribuem para ampliar seu polimorfismo hepático, entretanto pode ser considerado o ponto inicial para os estudos comparativos e filogenéticos entre os vertebrados. O fígado dos peixes aparece como em todos os outros vertebrados, como um órgão chave que vai controlar muitas funções vitais e realizar um papel proeminente na fisiologia dos peixes, tanto no anabolismo (proteínas, lipídios e carboidratos) e no catabolismo (nitrogênio, glicogenólises e desintoxicação). Por outro lado, deve ser considerado como um órgão alvo para muitos parâmetros biológicos e ambientais que podem alterar a estrutura e o metabolismo do fígado, como por exemplo, a alimentação, toxinas, parasitas, microrganismos e metais pesados acumulados. Nos peixes o fígado é localizado ventralmente na cavidade celomática, ajustando-se ao espaço disponível na cavidade do corpo. Foi realizada através da microscopia de luz e eletrônica de varredura a morfologia estrutural do desenvolvimento do fígado do tubarão azul nos 33 espécimes, divididos em diferentes tamanhos de embriões e fetos de 4 cm até 45 cm, comparados com um indivíduo adulto fêmea de 2 metros. A contagem de hepatócitos e vacúolos de gordura foi pela morfometria, através da técnica de pontos em fotomicrografias aleatórias. Nos resultados obtidos, pode-se notar que o fígado ocupava 20% do tamanho do animal. Microscopicamente, observou a presença de diferentes tamanhos de vacúolos de armazenamento de lipídio nos hepatócitos, diferença nas proporções de hepatócitos, linfócitos e vasos sanguíneos que diminui à medida que aumenta a estocagem de lipídios, consequentemente diminuindo a visibilidade da estrutura do fígado. Maior visualização de vacúolos translúcidos intracitoplasmáticos microgoticular aumentando gradativamente para macrogoticulares. Sugerindo assim que a presença de lipidios seja para manutenção dos filhotes, flutuabilidade e reserva energéticado animal, indicando que ele armazena gordura em seu fígado desde o início da embriogênese. / The blue shark (Prionace glauca), popularly known as blue dogfish among all shark species is the most abundant in the marine environment, and can be found in all seas. With the diversity of species, the description of any specific liver can hardly be used as a model. Together with this variability, some physiological characteristics of the fish contribute to increase its hepatic polymorphism, however, it can be considered the starting point for comparative and phylogenetic studies among vertebrates. Fish liver appears as in all other vertebrates as a key organ that will control many vital functions and play a prominent role in fish physiology, both in anabolism (proteins, lipids and carbohydrates) and in catabolism (nitrogen, glycogenolysis and detoxification). On the other hand, it should be considered as a target organ for many biological and environmental parameters that can alter the structure and metabolism of the liver, such as food, toxins, parasites, microorganisms and accumulated heavy metals. In fish, the liver is located ventrally in the coelomic cavity, adjusting to the available space in the body cavity. The structural morphology of blue shark liver development in 33 specimens divided into different sizes of embryos and fetuses from 4 cm to 45 cm was compared to a female adult of 2 meters, using light microscopy and scanning electron microscopy. The counts of hepatocytes and fat vacuoles were by morphometry, using the technique of points in random photomicrographs. In the results obtained, it can be noted that the liver occupied 20% of the size of the animal. Microscopically, it observed the presence of different sizes of lipid storage vacuoles in hepatocytes, a difference in the proportions of hepatocytes, lymphocytes and blood vessels that decreases as lipid storage increases, consequently decreasing the visibility of the liver structure. Greater visualization of microcyticular intracytoplasmic translucent vacuoles gradually increasing for macrogoticulares. Thus suggesting that the presence of lipids is for the maintenance of the puppies, buoyancy and energy reserve of the animal, indicating that it stores fat in its liver from the beginning of embryogenesis.

Due to an emergency status dune restoration project following Subtropical Storm Andrea in 2007 on Singer Island, Florida, nests needed to be moved during early to mid-incubation. Nesting success was compared between those moved mid-incubation, moved within 12h to either a native sand incubation area or a renourished sand incubation area, and those left in-situ. Nests moved within 12h to the native sand had a significantly larger proportion of pipped hatchlings. Nests moved mid-incubation had a significantly lower proportion of hatched eggs as well as emergent hatchlings. The stage in which embryonic development was arrested corresponded to the stage the embryos were in during the time of movement; indicating movement was the cause of death. When comparing nests moved within the initial 2.5 weeks of development to those moved after 2.5 weeks of development, there was no significant difference in hatching success. / by Natasha M. Ahles. / Vita. / Thesis (M.S.)--Florida Atlantic University, 2009. / Includes bibliography. / Electronic reproduction. Boca Raton, Fla., 2009. Mode of access: World Wide Web.

by Chan Wood-yee. / Thesis (Ph.D.)--Chinese University of Hong Kong, 1987. / Includes bibliographical references.

by Lam, Sau Hing. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1998. / Includes bibliographical references (leaves 141-158). / Abstract also in Chinese. / Content / Acknowledgements / Abbreviation / Project Objectives / Abstract / Chapter Chapter One --- Introduction / Chapter 1.1 --- The definition of homeobox genes / Chapter 1.2 --- Homeobox genes as a transcription factor / Chapter 1.3 --- Homeobox in Drosophila / Chapter 1.3.1 --- The development of Drosophila / Chapter 1.3.2 --- Maternal genes / Chapter 1.3.3 --- Segmentation genes / Chapter 1.3.4 --- Homeobox genes / Chapter 1.4 --- Family of Hox genes in the mammalian system / Chapter 1.5 --- Some possible chemical mechanism in the cascade system of the Hox genes in the vertebrate / Chapter 1.6 --- Hox (Antp-Class homeobox gene) in mammal / Chapter 1.6.1 --- Labial Like homeobox genes / Chapter 1.6.2 --- Proboscipedia Like homeobox genes / Chapter 1.7 --- Divergent homeobox genes / Chapter 1.7.1 --- Paired (prd) Class / Chapter 1.7.2 --- Even-Skipped (Eve) Class / Chapter 1.7.3 --- Distal-less (Dll) Class / Chapter 1.7.4 --- Muscle-Specific Homeobox (Msx) Class / Chapter 1.8 --- Orphan homeobox gene / Chapter 1.8.1 --- The characteristic of Hox 11 sequence in human and mouse / Chapter 1.8.2 --- Novel homeobox genes related to hox 11 gene family / Chapter 1.8.3 --- The mechanism of HOX 11 inducing gene regulation and signal transduction pathways / Chapter 1.8.4 --- HOX 11 in human / Chapter 1.8.5 --- Hox11 in mouse / Chapter 1.8.6 --- Hox11 L1 in mouse / Chapter 1.9 --- Homeobox gene involved in haematopoiesis / Chapter 1.10 --- Some translocations of homeobox genes in blood lineage / Chapter 1.11 --- The development of mouse / Chapter 1.11.1 --- Early organogenesis / Chapter 1.11.2 --- Nervous system development / Chapter 1.11.3 --- Somite development / Chapter 1.11.4 --- Eye development / Chapter 1.11.5 --- Neural crest cell migration / Chapter 1.11.6 --- Branchial arches development / Chapter Chapter Two --- Materials and methods / Chapter 2.1 --- Mouse Embryos / Chapter 2.2 --- RNA extraction / Chapter 2.3 --- Large plasmid preparation / Chapter 2.4 --- The synthesis of cDNAs using Reverse Transcription / Polymerase Chain Reaction (RT-PCR) and ligation into Bluescript® II KS / Chapter 2.4.1 --- The synthesis of RT-PCR products / Chapter 2.4.2 --- The formation of blunt ends of cDNA / Chapter 2.4.3 --- The ligation of cDNA with plasmid vectors / Chapter 2.4.4 --- Transformation / Chapter 2.4.5 --- The miniprep plasmid purification / Chapter 2.5 --- T7 sequencing / Chapter 2.6 --- Double stranded DNA cycle sequencing of plasmid / Chapter 2.6.1 --- Gel electrophoresis / Chapter 2.7 --- Northern blot / Chapter 2.7.1 --- Preparation of Northern blot / Chapter 2.7.2 --- Hybridization of Northern blot / Chapter 2.8 --- DIG labeled probes in whole mount in situ hybridization / Chapter 2.8.1. --- Preparation of linear DNA to generate riboprobes / Chapter 2.8.2 --- Preparation of DIG labeled riboprobe / Chapter 2.8.3. --- Preparation of embryo powder / Chapter 2.8.4 --- Absorption of antibody / Chapter 2.8.5 --- Embryo preparation / Chapter 2.8.6 --- Embryos staining / Chapter 2.9 --- Sections from whole mount in situ hybridization / Chapter 2.10 --- The radiolabeled section in situ hybridization / Chapter 2.10.1 --- The preparation of paraffin wax block and sample sections / Chapter 2.10.2 --- Slide pretreatment / Chapter 2.10.3 --- Preparation of probe / Chapter 2.10.4 --- Hybridization / Chapter 2.10.5 --- Washing / Chapter 2.10.6 --- Emulsification and development / Chapter 2.11 --- DIG-label in situ hybridization of frozen / Chapter 2.12 --- H&E staining / Chapter Chapter Three --- Results and Discussion / Chapter 3.1 --- Synthesis of Tlx3 probes for use in the section and whole mount in situ hybridization / Chapter 3.1.1 --- Synthesis of the Tlx cDNA by RT-PCR method / Chapter 3.1.2 --- The Tlx3 genomic clone for detecting the developmental expression of Tlx3 by Northern Blot / Chapter 3.1.3 --- The characterization of Tlx3 cDNAs and the sonic hedgehog cDNA / Chapter 3.2 --- Section in situ hybridization using different probes / Chapter 3.2.1 --- Section in situ hybridization using the transcribed riboprobes of Pax2 cDNA / Chapter 3.2.2 --- The transcribed riboprobes of Tlx genomic clones were used to hybridize the section in situ hybridization / Chapter 3.2.3 --- The in situ hybridization of frozen sections of mouse embryos using the transcribed riboprobes from Tlx3 cDNA subclone / Chapter 3.3 --- Expression pattern of Tlx3 on whole mount embryos using both cDNA and genomic probes / Chapter 3.3.1 --- The expression of Tlx3 on whole mount in situ hybridization of the mouse embryos using the antisense probes from the Tlx3 genomic clone / Chapter 3.3.2 --- Whole mount in situ hybridization of mouse embryo using the transcribed riboprobes of Tlx3 cDNA / Chapter 3.3.3 --- The expression pattern of sonic hedgehog on embryos at 8.5 to9.5 dpc / Conclusion / Future prospective / Appendix / Reference / Acknowledgement

by Tse Kam Wah. / "September 1998." / Thesis (Ph.D.)--Chinese University of Hong Kong, 1998. / Includes bibliographical references (p. 123-145). / 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.

Forkhead domain基因家族編碼了很多對於胚胎發育至關重要的轉錄因子，而Foxp4則屬於p-subtype forkhead轉錄因子其中一員。Foxp4在胚胎發育期間的表達十分活躍，在發育中的腦部的不同地方表達，但其於中樞神經系統發育中的調控角色並不清楚。Foxp4基因剔除小鼠在出生前死於心臟的缺陷表型(心二分支) ，在此時間段，腦部的發育才剛剛開始，因此我們無法利用Foxp4基因剔除小鼠作為研究中樞神經系統發育的動物模型。最近，我們的團隊利用小腦組織培養技術及siRNA發佈的研究顯示，Foxp4在小鼠小腦中的蒲金氏細胞(Purkinje cell)中擔當著重要的維持作用。這項研究結果加深了我們對研究Foxp4在中樞神經系統發育中的調控角色的決心。 / 本論文旨在利用斑馬魚作為實驗模型，研究foxp4在斑馬魚中樞神經系統發育中的表達及調控角色。RT-PCR結果顯示foxp4在斑馬魚發育中的bud stage開始表達，並在及後的階段維持其表達水平。利用原位雜交技術 (whole mount in-situ hybridization)，我們發現foxp4表達的地區主要集中於發育中的腦部。在成年斑馬魚中，foxp4表達在不同組織和器官，包括腦部，眼睛和心臟。成年斑馬魚腦部切片原位雜交 (sectioned in-situ hybridization)則顯示，foxp4在小腦的蒲金氏細胞和視頂蓋(optic tectum)的periventricular gray zone表達。 / 為了進一步探究foxp4對於胚胎發育過程中的功能，我們利用微注射技術，把反義嗎啉 (morpholino) MO1注射到斑馬魚胚胎中，大幅度抑制foxp4的表達水平。胚胎受精後48小時，MO1注入的胚胎顯示出第四腦室腦積水的缺陷表型。組織學分析顯示，第四腦室以下的延髓被壓縮致形態異常。此外，利用原位雜交技術及不同的分子標記，我們發現胚胎的中後腦邊界也會出現輕度畸形，而後腦的神經元數量及排列亦受到影響。 / 本項研究展示foxp4在胚胎中樞神經系統的發展的重要性，亦提供了新的見解。我們認為foxp4可能是調控腦室發育的重要成員，但在此方面與foxp4相關的分子機制仍須作更深入的研究。 / The forkhead domain gene family encodes a large group of transcription factors that play essential roles in development. Foxp4 is one of the members in the Foxp subfamily that expressed in different parts of developing central nervous system (CNS) and its function is less characterized. Previous study on Foxp4-knockout mice resulted in early embryonic lethality due to defective heart tube development that hindered the functional study of Foxp4 in CNS development. Recently, our laboratory reported that Foxp4 functions as a maintenance role in the Purkinje cell in the mouse cerebellum. Nevertheless, the role of foxp4 in CNS development was still unclear. / In this study, we used zebrafish as a model to study the expression pattern and functional study of foxp4 in the developing CNS. RT-PCR analysis showed that foxp4 transcript was expressed at the bud stage and maintained in the later embryonic stages. Whole-mount in-situ hybridization showed that foxp4 expressed in the cephalic region during embryonic development. In adult zebrafish, foxp4 expresses in different tissues and organs including brain, eye and heart. Sectioned in-situ hybridization of the adult zebrafish brain showed that foxp4 was specifically expressed in the Purkinje cell and the periventricular gray zone of optic tectum. / To further investigate the function of foxp4 during embryonic development, we injected antisense morpholino, MO1 into the zebrafish embryo to knockdown foxp4. By 48 hour post fertilization (hpf), MO1-injected embryos displayed hydrocephalus in the 4th ventricle. Histological analysis revealed that the medulla oblongata below the 4th ventricle was compressed by the edema resulting in abnormal morphology of medulla oblongata in the MO1-injected morphant. In addition, a mild malformation of the mid-hindbrain boundary, disrupted hindbrain patterning was observed in MO1-injected morphant. / Our findings provide new insight into the function of foxp4 in embryonic CNS development. We suggested that foxp4 may be essential in regulating the brain ventricle development while the molecular mechanism underlying the functional role of foxp4 requires further investigation. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Wong, Wai Kei. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 92-102). / Abstracts also in Chinese. / Abstract --- p.iii / 摘要 --- p.v / Acknowledgement --- p.vii / Figure and table list --- p.xi / Abbreviation --- p.xii / Chapter Chapter 1 --- General Introduction / Chapter 1.1 --- Zebrafish as a developmental model --- p.1 / Chapter 1.2 --- Zebrafish development with highlights --- p.3 / Chapter 1.2.1 --- CNS development --- p.3 / Chapter 1.3 --- Forkhead domain gene in development --- p.5 / Chapter 1.3.1 --- History of forkhead domain gene --- p.5 / Chapter 1.3.2 --- Functional roles of forkhead domain genes in development --- p.6 / Chapter 1.4 --- Foxp subfamily --- p.8 / Chapter 1.4.1 --- Diverse functions of Foxp1, 2, 3 and 4 --- p.8 / Chapter 1.4.2 --- Relationship between Foxp subfamily members --- p.10 / Chapter 1.5 --- Foxp4 --- p.11 / Chapter 1.5.1 --- Genomic organization and protein structure of mFoxp4 --- p.11 / Chapter 1.5.2 --- Previous studies of mFoxp4 --- p.14 / Chapter 1.5.3 --- Foxp4 studies in other model organisms --- p.14 / Chapter 1.5.3.1 --- Rat --- p.15 / Chapter 1.5.3.2 --- Xenopus --- p.16 / Chapter 1.5.3.3 --- C. elegans --- p.16 / Chapter 1.5.4 --- Zebrafish foxp4 --- p.17 / Chapter 1.5.5.1 --- Genomic organization and protein structure of foxp4 --- p.17 / Chapter 1.5.5.2 --- Sequence alignment of foxp4 with other models --- p.19 / Chapter 1.6 --- Hypothesis, aim and strategy of the study --- p.22 / Chapter Chapter 2 --- Expression of foxp4 in zebrafish embryo and adult zebrafish brain / Chapter 2.1 --- Introduction --- p.24 / Chapter 2.2 --- Materials and methods --- p.25 / Chapter 2.2.1 --- Animals --- p.25 / Chapter 2.2.2 --- Materials --- p.26 / Chapter 2.2.3 --- Semi-quantitative PCR --- p.35 / Chapter 2.2.3.1 --- cDNA of zebrafish embryo --- p.35 / Chapter 2.2.3.2 --- Isolation of adult zebrafish organs --- p.36 / Chapter 2.2.3.3 --- RNA extraction and reverse transcription --- p.36 / Chapter 2.2.3.4 --- Polymerase chain reaction --- p.37 / Chapter 2.2.4 --- Subcloning of DNA fragment / Chapter 2.2.4.1 --- Preparation of cloning vectors --- p.40 / Chapter 2.2.4.2 --- Subcloning of DNA fragments --- p.40 / Chapter 2.2.4.3 --- Transformation of DNA into competent cells --- p.40 / Chapter 2.2.4.4 --- Preparation of recombinant plasmid DNA --- p.41 / Chapter 2.2.5 --- Whole mount in-situ hybridization of zebrafish embryo --- p.45 / Chapter 2.2.5.1 --- Preparation of equipment --- p.45 / Chapter 2.2.5.2 --- Preparation of zebrafish embryos --- p.45 / Chapter 2.2.5.3 --- Preparation of RNA probe --- p.46 / Chapter 2.2.5.4 --- Whole-mount in-situ hybridization --- p.48 / Chapter 2.2.6 --- Sectioned in-situ hybridization of adult zebrafish brain --- p.49 / Chapter 2.2.6.1 --- Histology of adult zebrafish brain --- p.49 / Chapter 2.2.6.2 --- Sectioned in-situ hybridization --- p.50 / Chapter 2.3 --- Results --- p.51 / Chapter 2.3.1 --- Expression profile of foxp4 in different stages of zebrafish embryo --- p.51 / Chapter 2.3.2 --- Expression pattern of foxp4 in different stages of zebrafish embryo --- p.54 / Chapter 2.3.3 --- Expression profile of foxp4 in different zebrafish organs and tissues --- p.57 / Chapter 2.3.4 --- Expression pattern of foxp4 in adult zebrafish brain --- p.59 / Chapter 3.4 --- Discussion --- p.61 / Chapter Chapter 3 --- Functional analysis of foxp4 in zebrafish embryonic development / Chapter 3.1 --- Introduction --- p.63 / Chapter 3.2 --- Materials and methods --- p.64 / Chapter 3.2.1 --- Materials --- p.64 / Chapter 3.2.2 --- Design of morpholino --- p.68 / Chapter 3.2.3 --- Sequencing of morpholino target regions of foxp4 --- p.70 / Chapter 3.2.4 --- Microinjection --- p.70 / Chapter 3.2.4.1 --- Preparation of materials and equipment --- p.70 / Chapter 3.2.4.2 --- Preparation of injection needle --- p.70 / Chapter 3.2.4.3 --- Preparation of morpholinos --- p.70 / Chapter 3.2.4.4 --- Calibration of injection volume --- p.71 / Chapter 3.2.4.5 --- Microinjection of zebrafish embryo --- p.71 / Chapter 3.2.5 --- Western blotting to assay foxp4 translation inhibition --- p.72 / Chapter 3.2.5.1 --- Preparation of protein extracts --- p.72 / Chapter 3.2.5.2 --- Coomassie blue staining --- p.73 / Chapter 3.2.5.3 --- Western blotting --- p.74 / Chapter 3.2.6 --- Whole mount in-situ hybridization --- p.74 / Chapter 3.3 --- Results --- p.75 / Chapter 3.3.1 --- MO1 knockdown efficiency assayed by Western blotting --- p.75 / Chapter 3.3.2 --- General morphology of morphants --- p.77 / Chapter 3.3.3 --- Histology at the hindbrain region showing the phenotype --- p.79 / Chapter 3.3.4 --- Whole mount in-situ hybridization of different molecular markers --- p.81 / Chapter 3.4 --- Discussion --- p.85 / Chapter Chapter 4 --- Future directions and conclusion / Chapter 4.1 --- Future directions --- p.89 / Chapter 4.2 --- Conclusion --- p.91 / Reference --- p.92