Dissertation > Agricultural Sciences > Aquaculture, fisheries > Aquatic basic science > Aquatic Biology > Aquatic Zoology

Studies on the Main Functioning Cytology in the Intestine, Spleen and Pituitary Gland of Chinese Soft-Shelled Turtle, Pelodiscus Sinensis

Author BaoHuiJun
Tutor ChenQiuSheng
School Nanjing Agricultural College
Course Basic Veterinary
Keywords Soft-shelled Turtle Intestinal Spleen Pituitary Cytology
CLC S917.4
Type PhD thesis
Year 2010
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The soft-shelled turtle is important aquaculture animal has a high nutritional value and medicinal value. In biology, soft-shelled turtle belonging Reptilia, has the characteristics of the hibernation phenomenon and variable temperature, digestion, immune and endocrine and other physiological activities and metabolic processes are different from those of other animals. With our captivity scale expanding, soft-shelled turtle popular and spread of infectious diseases is more and more serious. In the past, the more test study on soft-shelled turtle are concentrated in the field of disease-related, the lack of basic research to its special structure, the data were compared. In this paper, the soft-shelled turtle with Chinese characteristics as a model animal, a detailed study of its representative organs and tissues: cytologic features of the digestive system of the gut, spleen of the immune system, the endocrine system, the pituitary gland, and analysis of its functional significance . The thesis includes turtle the intestinal mucosa mechanical barrier and immune barrier cell composition and structural characteristics; interstitial cells of Cajal in intestinal distribution differences and ultrastructural characteristics; spleen ellipsoid endothelial capillaries discovery and cell lymphocyte migration Foundation; anterior pituitary endocrine the cell ultrastructure its seasonal changes. From the above three aspects of the biological characteristics and significance of soft-shelled turtle, laid the theoretical foundation for further study its physiological function, immune function, or hibernating and non-hibernating physiological cycle, as well as artificial breeding, disease prevention and treatment to provide a scientific basis. Experiment Ⅰ China turtle intestinal epithelial the mucosal barrier structure with cells characterized by light and electron microscopy under normal circumstances, the structural features of the soft-shelled turtle intestinal epithelial mucosal barrier, statistical analysis the main component of the intestinal immune barrier of soft-shelled turtle in different months - intraepithelial lymphocytes (IEL) in the distribution pattern of the small intestine and the large intestine. The results showed that: the intestinal mucosa of simple columnar or pseudostratified columnar epithelium. Tightly packed between the top of the epithelial cells of the mucosal epithelium, developed tight junctions, the intermediate connection desmosomes cell junctional complex. Desmosomes and the fitting is also exists between the middle of the mucosal epithelium and the epithelial cells of the base. Most of the cytoplasm of epithelial cells of intestinal mucosa contains a small amount of mitochondria, a small amount of epithelial cell cytoplasm rich in mitochondria and lipid droplets. Colorectal mucosa epithelium pseudostratified columnar epithelium, epithelial cytoplasmic lipid droplets were no distribution. The top of the epithelial cells of the mucosal epithelium tightly packed, central and basal epithelial cells arranged loosely, the gap between the cells are large and obvious. Many mucus cells scattered between distributed throughout the intestinal mucosal epithelial cells, mucus in some cytoplasmic granules electron density is low, and some high electron density. Mucus secreted by the mucous cells covering the surface of the intestinal mucosa. The intestinal epithelial lack Microfole cells. Intestinal mucosal epithelial cells between the diameter of a 1-7gm channel, sometimes visible lymphocyte distribution channel. IEL the IEL (diameter 4-9μm) is located between the intestinal mucosal epithelial cells extend pseudopodia, and between them, showing migration trends. The intestine visible complete free lymphocytes. Central and base the distribution of plasma cells, usually found between the epithelial cells in the intestinal mucosal epithelial plasma cells in two forms: a kind of rough endoplasmic reticulum for the expansion of the state, another flat cystic. IEL cell number gradually decreased from May, September to December. The above test results suggest that the turtle intestinal mucosal epithelial mucosal barrier function. Epithelial cells and its tight junctional complex, mucus secreted by the mucous cells constitute a mechanical barrier; plasma cells, IEL and channel constitute the immune barrier. Turtle intestinal mucosal immune cell distribution is also affected by season. CD3 T II soft-shelled turtle intestinal mucosal immune barrier test, the distribution of CD22 B lymphocytes by immunocytochemistry SABC method to observe the soft-shelled turtle intestinal immune barrier CD3 T CD22 B lymphocytes distribution characteristics, statistics and analysis of soft-shelled turtle bowel CD3 T lymphocytes, CD22 B lymphocyte distribution law. The results showed that: CD3 T lymphocytes are mainly distributed in the intrinsic membrane, a small amount found in the mucosal epithelium. Majority of CD22 B lymphocytes in the mucosal epithelium, the intrinsic membrane is rare. Intraepithelial, CD3 T lymphocytes in the small intestine from front to back, in ascending order, the largest number of the rear section of the small intestine; descending order in the large intestine of CD3 T lymphocytes. Intrinsic membrane, CD3 T lymphocytes in the small intestine, the middle of the highest density in the large intestine descending order. Intraepithelial intrinsic membrane CD3 T lymphocytes each bowel compared, a significant difference between most of the bowel. The number of lamina propria CD3 T lymphocytes intraepithelial 3.11 times the number of the large intestine, small intestine CD3-T lymphocytes. CD22 B lymphocytes in terms of epithelial, CD22 B lymphocytes in the largest number in the posterior segment of the small intestine, and in the large intestine gradually reduced from front to back; lamina propria, CD22 B lymphocytes from the small intestine to the posterior segment of the small intestine in turn reduce in turn reduce the large intestine. Each bowel intraepithelial CD22 B lymphocytes compared to most of the bowel was no significant difference between the intrinsic membrane CD22 B lymphocytes were no significant differences between each bowel. The number of intraepithelial CD22 B lymphocytes of the lamina propria of 7.99 times the number of the small intestine of CD22 B lymphocytes is 1.75 times that of the large intestine. The above test results indicate that the distribution of T, B lymphocytes in the immune barrier turtle intestinal paragraphs differences, suggesting that they play a different biological role in the immune response in the intestinal mucosa. Soft-shelled turtle, test Ⅲ intestinal S-100 protein immunocytochemistry reaction study using immunohistochemistry technology, S-100 protein-positive cells in the soft-shelled turtle distribution pattern of the intestinal tract. The results show: S-100 protein-positive cells can be divided into two types: the first type of S-100 positive cells irregular the cell protrusions thickness varying lengths, mainly located in the small intestine lamina propria and submucosa, large intestine almost no positive cell distribution. The second type of S-100 protein-positive cells divided into bipolar and astrocytes. Bipolar S-100 protein-positive cell processes and some long, some short. They are distributed in the intestine and colon of the submucosa and circular muscle between annular intramuscular, circular muscle and between the longitudinal muscle of the large intestine submucosa. Star S-100 protein-positive cells were only distributed in the lamina propria of the large intestine preceding paragraph, these cells contain a plurality of protrusions, intertwined to form a network-like structure between the protrusions. S-100 protein is a dendritic cell-specific antibodies, but is also expressed in the interstitial cells of Cajal Division, glial cells. A type of S-100 protein-positive cells in this experiment as dendritic cells, according to the shape and location of the second type of S-100 protein-positive cells in interstitial cells of Cajal or glial cells. Studies suggest that dendritic cells with antigen-presenting role of interstitial cells of Cajal the intestinal peristalsis pacemaker, glial cells can maintain the integrity of the intestinal epithelial. Therefore, it is speculated that dendritic cells involved in the immune barrier function in the turtle the small intestine, the interstitial cells of Cajal and glial cells, respectively regulate bowel movement and intestinal barrier function. Test Ⅳ Cajal interstitial cells in China the turtle gut the distribution differences and ultrafine characteristics this paper, c-Kit immunocytochemistry technology and transmission electron microscopy to study the distribution of the interstitial cells of Cajal (ICC) in the intestinal tract of soft-shelled turtle and ultrastructural features . c-Kit chemical reaction of the immune cells, interstitial cells of Cajal divided into bipolar and astrocytes. Bipolar c-Kit the ICC cell processes and some long, some short or obvious. The distribution of these cells in the large intestine submucosa, between the small and large intestine submucosa and circular muscle, between the annular intramuscular circular muscle and longitudinal muscle, serosa. In the submucosa, bipolar c-Kit ICC distribution around the blood vessels. Submucosa and circular muscle between the circular muscle and longitudinal muscle, showing the distribution of linear or in clusters, sometimes around the blood vessels around. Intramuscular, bipolar c-Kit ICC around the blood vessels around or parallel to the smooth muscle cell distribution in the annular. In serous bipolar c-Kit ICC scattered. Star c-Kit ICC cells only distributed intrinsic membrane in the large intestine, these cells have a plurality of projections, between the projections intertwined to form a mesh structure, mainly around in the perivascular distribution. Electron microscope, the the bipolar and star ICC cytoplasm contains mitochondrial membrane invagination vesicles, rough endoplasmic reticulum and more irregular cytoplasmic projections. Between ICC cells and ICC cells, gap junctions between cells and nerves of the ICC, ICC cells and smooth muscle cells. The above test results suggest that the soft-shelled turtle intestinal ICC cells has a unique distribution characteristics and ultrastructure, its morphological characteristics and its consistent with the important role in the maintenance of intestinal motility. Test China turtle spleen ellipsoid V high endothelial capillaries found cytological basis of this article and lymphocyte migration by light and electron microscopy technology, the organizational structure of the soft-shelled turtle spleen white pulp, the soft-shelled turtle spleen ellipsoid vascular other animals, their blood vessels are high endothelial capillary endothelial cells were high columnar or cuboidal, equivalent to mammalian high endothelial venules. Spleen memory the blood - spleen screen structure, on the one hand can prevent foreign antigen (carbon particles) into the ellipsoid peripheral lymphoid sheath (PELS), on the other hand may be the formation of a special temporary channel to allow lymphocyte migration in which to reach PELS. passage structure is made of reticular cells, reticular fibers, migrated cells (lymphocytes, macrophages, RBC), support cells, together constitute a composite structure of the ellipsoid related cells, channel. They form a microenvironment conducive lymphocyte movement. The test found that the the lymphocyte migration process is as follows: the channel is formed between the ellipsoid high the endothelial capillary endothelial cells, lymphocytes along the passage through the the ellipsoid endothelial capillary basement membrane to reach the ellipsoid; lymphocytes in Sertoli cells and its protruding effect along the channel, from the ellipsoid to reach the PELS outside; lymphocytes ellipsoid cells and their projections linked PELS PELS outside to enter. The above test results lymphocyte homing cytological basis for in-depth study of reptiles. Test Ⅵ endocrine cells of the soft-shelled turtle adenohypophysis ultrastructure and seasonal changes using transmission electron microscopy or hibernating and non-hibernating soft-shelled turtle adenohypophysis in various endocrine cell ultrastructure observed. The results show, the adenohypophysis mainly there are five types of endocrine cells: cells of growth hormone (GH), prolactin (PRL) cells, the cells of the thyroid stimulating hormone (TSH), promoting the adrenal cortex hormone (ACTH) cells and gonadotropin (GTH) cells. . They have their ultrastructural features: GH cells were round or oval-shaped, 10-12μm in diameter; distribution of secretory granules, dense rounded. PRL cells oval or irregular in shape, diameter 7-9μm nucleus irregular shape, the cytoplasm of high electron density. TSH cells were oval or irregular polygonal, diameter 8-12μm, oval nuclei, secretory granules in small quantities, are mainly distributed in the cell periphery. ACTH cells irregular diameter of 8-10μm, and the nucleus is oval, less secretory granules. GTH cells round or oval, diameter 12-15μm, the nucleus is round or oval, cytoplasm, large, small two secretory granules. Meanwhile, the turtle adenohypophysis of the structural features of the endocrine cells and secretory granules density existence of certain seasonal changes. Hibernation period, GH cytoplasmic organelles than non-hibernating significantly reduced; TSH cells is higher than the number of non-hibernating period of hibernation; the hibernation period GTH cells within the secretory granules appear larger fusion phenomenon, particle diameter was significantly greater than the non- period of hibernation, the largest up to 2μm. The PRL cells ACTH cell ultrastructure no obvious seasonal variation.

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