From molecules to organs : Microscopy and multi-scale nature of development

Morphogenesis is a phenomenon of intricate balance and dynamic interplay between processes occurring at a wide range of scales (spatial, temporal and energetic). During development, a variety of physical mechanisms are employed by tissues to simultaneously pattern, move, and differentiate based on information exchange between constituent cells, perhaps more than at any other time during an organism's life. To fully understand such events, a combined theoretical and experimental framework is required to assist in deciphering the correlations at both structural and functional levels at scales that include the intracellular and tissue levels as well as organs and organ systems. Microscopy, especially diffraction-limited light microscopy, has emerged as a central tool to capture the spatio-temporal context of life processes. Imaging has the unique advantage of watching biological events as they unfold over time at single-cell resolution in the intact animal. In this work I present a range of problems in morphogenesis, each unique in its requirements for novel quantitative imaging both in terms of the technique and analysis. Understanding the molecular basis for a developmental process involves investigating how genes and their products- mRNA and proteins-function in the context of a cell. Structural information holds the key to insights into mechanisms and imaging fixed specimens paves the first step towards deciphering gene function. The work presented in this thesis starts with the demonstration that the fluorescent signal from the challenging environment of whole-mount imaging, obtained by in situ hybridization chain reaction (HCR), scales linearly with the number of copies of target mRNA to provide quantitative sub-cellular mapping of mRNA expression within intact vertebrate embryos. The work then progresses to address aspects of imaging live embryonic development in a number of species. While processes such as avian cartilage growth require high spatial resolution and lower time resolution, dynamic events during zebrafish somitogenesis require higher time resolution to capture the protein localization as the somites mature. The requirements on imaging are even more stringent in case of the embryonic zebrafish heart that beats with a frequency of ~ 2-2.5 Hz, thereby requiring very fast imaging techniques based on two-photon light sheet microscope to capture its dynamics. In each of the hitherto-mentioned cases, ranging from the level of molecules to organs, an imaging framework is developed, both in terms of technique and analysis to allow quantitative assessment of the process in vivo. Overall the work presented in this thesis combines new quantitative tools with novel microscopy for the precise understanding of processes in embryonic development.

Glycocalyx production in teleosts [Translation from: Verhandlungen der Deutschen Zoologischen Gesellschaft, p.286, 1970]

Shielding the organism against harmful effects from the environment is one of the most important tasks of the outer covering of all animals. The epidermis of primarily aquatic organisms and the epithelia of organs which are exposed to water, such as the digestive or the urinary system, possess a film of glycoproteins and mucopolysaccharides, the glycocalyx. This short paper examines the relationship of the mucus cells with the glycocalyx.

Redclaw crayfish aquaculture in Ecuador: the new boom?

Introduction of red claw crayfish (Cherax quadricarinatus) in Ecuador is presented together with the status and future of crayfish culture in the country.

Crayfish survey of the Weaver, Dane, Goyt and Etherow catchments

This is the report on the Crayfish Survey of the Weaver, Dane, Goyt and Etherow catchments from 1998 by the Environment Agency. The aims of this report are: Firstly, to present the findings of the crayfish survey and details of the sites visited. Secondly to present the information on distribution maps with past records so that the current status can be seen and finally to use this information so that recommendations for the conservation of native crayfish can be made in accordance with the national action plan for this species and the Environment Agency’s Species Management Programme. The report contains sections on background, going through legislation, distribution and requirements of both native and non-native crayfish. Sections on methodology, results and discussion, conclusion and recommendations. The appendix I contains maps showing the sampling points locations. Details of sampling sites are summarized in appendix II. Appendix III contains previous crayfish records and Appendix IV shows the field data recording form. Finally, a collection of photographs are displayed in appendix V.

Additional crayfish survey of Checkley Brook, Hollywood End Brook and Black Brook (Goyt) 1999

This is the report on the Additonal Crayfish Survey of Checkley Brook, Hollywood End Brook and Black Brook from 1999 by the Environment agency. The aim of the 1999 survey was to obtain a more complete picture of the crayfish distribution in those areas. It contains sections on the sampling methodology which followed the sampling done in 1998, the results of the sampling indicating the species of crayfish occurring in the sampling areas and some discussion and conclusions for each area. The appendix I contains maps locating the sampling points and past records. The appendix II contains detailed information of the sampling points.

River Goyt and Etherow Crayfish Survey

This is the River Goyt & Etherow Crayfish Survey report from the Environment Agency held between August and November 2000. The report focuses on the need to verify the presence of the non-native American signal crayfish (Pacifastacus leniusculus) in the main River Goyt, and determine its distribution. Signal crayfish have been reported in the River Goyt after spreading from a known population centre in Hollywood End Brook, to the east of Marple Bridge. The report contains sections on results, discussion and a summary. The section on results shows maps of presence or absence of different crayfish species in River Goyt, and the Mersey/ Weaver river systems such American signal crayfish (Pacifastacus leniusculus) and European freshwater crayfish (Austopotamobius pallipes).

Aphanomycosis of crayfish: crayfish plague

This is the Aphanomycosis of crayfish: crayfish plague report produced by the Environment Agency in 2000. Crayfish plague is an extremely virulent fungal disease of European crayfish species, the white clawed or stone crayfish of Western Europe Austropotamobius pallipes, the Noble crayfish of northern Europe Astacus astacus and the narrow clawed crayfish of Eastern Europe, Astacus leptodactylus. The white claw crayfish A. pallipes is the indigenous native crayfish of the British Isles. Until the early 1980s there were extensive healthy populations of this crayfish in almost all suitable alkaline river and lake environments in England and Wales as far north as Northumberland. The conservation importance of this native crayfish is widely recognised. This report provides a general review of the literature of crayfish plague, including an overview of its spread through the British Isles from CEFAS records. Information on current diagnostic methods from the Office International des Epizooties (OIE) Aquatic Disease Manual is provided. Information on the taxonomy, morphology and physiology of the pathogen is reviewed, together with the pathogenicity and pathology of the disease and current means of prevention and control.

Studies on the filtration, feeding and excretion rates in Perna viridis, Marcia cor and Crassostrea gryphoides (Mollusca: Bivalvia) using P³² labelled Ankistrodesmes

The study deals with a series of experiments to investigate feeding and excretion in three species of bivalves: Perna viridis (Linné), Marcia cor (Sowerby) and Cassostrea gryphoides (Gould) from Manora Channel, Karachi. Bivalves were fed with suspensions of Ankistrodesmes labelled with P³². These animals showed a considerable variation in the average filtration rates depending upon species and the body lenght. Exceptionally high content of the P³² introduced with Akistrodesmes, got excreted as pseudofaeces and faeces within first three days following its absorption as a meal. The assimilated P³² is partly released as faecal material and its major proportion is directly transferred to the solution. As expexted the gonad and kidney are the main organs found responsible for excretion as comared to other body parts. Although, the assimilated P³² is mostly concentrated in the digestive glands, the results also show a significant presence of P³² in the gonads. Accumulation of P³² was the least in the foot.

A study of the male reproductive organs of nine species of penaeid prawns (Crustacea: Penaeinae)

This paper deals with the male reproductive organs of nine species of penaeid prawns; Penaeus penicillatus Alcock, P. merguiensis De Man, P. semisulcatus De Haan, Metapenaeus affinis (H.Milne Edwards), M. monoceros (Fabricius), M. stebbingi Nobili, Parapenaeopsis hardwickii (Miers), P. sculptilis (Heller) and P. stylifera (H. Milne Edwards). The male reproductive organs exhibited structural variations, which were more pronounced at generic level. These variations are mainly due to the type of spermatophore they possess. One species of each genus, that is, P. merguiensis, M. affinis and P. sculptilis were also studied histologically to examine the internal structure of the male reproductive organs. Spermatophores of the six species belonging to the genera Penaeus and Parapenaeopsis are also described and illustrated.

Bacterial load in pond water and different organs of a Indian major carp Cirrhinus mrigala Ham.

During the study period (August, 1993 to July,l994) the mean bacterial load in surface water was found to vary from 1.39 xl05 (July'94) to 3.llxl07CFU/ml (September'93), while that of botrom water ranged from l.Olxl06 (May'94) to 5.90xl07CFU/ml (October '93). The mean total number of bacterial load in body slime, liver and kidney was found to vary from 0.58xl03 (July'94) to 2.37xl07CFU/g (March'94),from 0.22xl03(July'94)to 9.64xl06 CFU/g (March'94) from O.l5xl03 (July'94) to 9.36xl06 CFU/g (March'94), respectively. Bacterial load in slime was significantly correlated with bacterial load in liver, bacterial load in slime was significantly correlated with bacterial load in kidney and bacterial load in liver was significantly correlated with bacterial load in kidney.