Gut-associated lymphoid tissue (GALT) of the goldfish,Carassius auratus

The head kidney and spleen are major sites of haemopoiesis in fish; a secondary center is found in loose connective tissue of the intestine. In this study we determined the nature of gut-associated haemopoietic tissue in the goldfish, Carassius auratus, using light and electron microscopy. This tissue is a loose stroma of reticular cells and fibers vascularized by capillaries, venules, and arterioles. The cellular population includes lymphoblasts, small and medium-sized lymphocytes, plasmocytes, macrophages, and various granulocytes. The most abundant granulocyte is the mast cell, whose large granules stain with Alcian blue and toluidine blue. Heterophils are found in the intestinal connective tissue as well as two other granulocytes: one with ovoid granules having dense parallel lamellae and another with granules containing crystalline inclusions. Immature forms of both granulocytes were also noted. Macrophages containing phagocytosed debris were often located close to the epithelium; they were observed forming clusters with lymphocytes. The epithelium contained a number of migrating leucocytes including lymphocytes and lymphoblasts, macrophages, and heterophils. Although many granulocytes were found in the connective tissue, granulopoiesis does not seem to be a major function. Gut-associated haemopoietic tissue in goldfish resembles diffuse lymphoid tissue and may be involved in intestinal immune responses.     

Antibodies against pax6 immunostain amacrine and ganglion cells and neuronal progenitors,but not rod precursors,in the normal and regenerating retina of the goldfish

Pax6 is a developmental regulatory gene that plays a key role in the development of the embryonic brain, eye, and retina. This gene is also expressed in discrete groups of neurons within the adult brain. In this study, antibodies raised against a fusion protein from a zebra fish pax6 cDNA were used to investigate the expression of the pax6 gene in the mature, growing, and regenerating retina of the goldfish. On western blots of retinal proteins, the pax6 antibodies recognize a single band at the approximate size of the zebra fish pax6 protein. In retinal sections, the antibodies label the nuclei of mature amacrine and some ganglion cells. At the retinal margin, where neurogenesis and cellular differentiation continually occur in goldfish, the antibodies label neuronal progenitors and the newly postmitotic neurons. Following injury and during neuronal regeneration, the antibodies label mitotically active progenitors of regenerating neurons. Rod precursors, proliferating cells that normally give rise solely to rod photoreceptors and are the presumed antecedents of the injury-stimulated neuronal progenitors, are not immunostained by antibodies to the pax6 protein. The results of this study document the identity of pax6-expressing cells in the mature retina and demonstrate that in the goldfish pax6 is expressed in neuronal progenitors during both retinal growth and regeneration. © 1996 John Wiley & Sons, Inc.     

Inflammatory response to Dentitruncus truttae (Acanthocephala) in the intestine of brown trout

Brown trout, Salmo trutta L., were infected with the acanthocephalan Dentitruncus truttae with the most affected areas being the anterior (near the pyloric caeca) and middle intestine. The parasite attached with a proboscis which usually penetrated the mucosa, lamina propria, stratum compactum, stratum granulosum and, sometimes, the muscularis layer. Around the parasite's body was an area of inflammatory tissue. At the point of attachment the lamina propria was thickened and the stratum compactum, stratum granulosum and muscularis layer were disrupted by proboscis penetration. Rodlet cells were more numerous in infected fish (P<0.01), and were found in the epithelial layer away from the worm. Infected intestines had larger numbers of mast cells (P<0.01), often in close proximity to, and inside, the blood capillaries and associated with fibroblasts of the muscularis layer and the stratum granulosum. Their migration toward the site of infection was suggested. Intense degranulation of mast cells was encountered in all intestinal layers especially near the parasite's body. Immunohistochemical tests were conducted on sections of intestinal tissue of uninfected and infected fish revealing the presence of met-enkephalin and serotonin (5-HT) in immuno-related cells of the intestine wall. Infected trout had larger numbers of elements positive to met-enkephalin and serotonin antisera. These data provided evidence for the role of the immune system of brown trout in the modulation of the inflammatory response to D. truttae. Results are discussed with respect to host immune response to an intestinal helminth.     

Invasion of microglia/macrophages and granulocytes into the Mauthner axon myelin sheath following spinal cord injury of the adult goldfish,Carassius auratus

Rapid activation of resident glia occurs after spinal cord injury. Somewhat later, innate and adaptive immune responses occur with the invasion of peripheral immune cells into the wound site. The activation of resident and peripheral immune cells has been postulated to play harmful as well as beneficial roles in the regenerative process. Mauthner cells, large identifiable neurons located in the hindbrain of most fish and amphibians, provided the opportunity to study the morphological relationship between reactive cells and Mauthner axons (M-axons) severed by spinal cord crush or by selective axotomy. After crossing in the hindbrain, the M-axons of adult goldfish, Carassius auratus, extend the length of the spinal cord. Following injury, the M-axon undergoes retrograde degeneration within its myelin sheath creating an axon-free zone (proximal dieback zone). Reactive cells invade the wound site, enter the axon-free dieback zone and are observed in the vicinity of the retracted M-axon tip as early as 3 hr postinjury. Transmission electron microscopy allowed the detection of microglia/macrophages and granulocytes, some of which appear to be neutrophil-like, at each of these locations. We believe that this is the first report of the invasion of such cells within the myelin sheath of an identifiable axon in the vertebrate central nervous system (CNS). We speculate that microglia/macrophages and granulocytes that are attracted within a few hours to the damaged M-axon are part of an inflammatory response that allows phagocytosis of debris and plays a role in the regenerative process. Our results provide the baseline from which to utilize immunohistochemical and genetic approaches to elucidate the role of non-neuronal cells in the regenerative process of a single axon in the vertebrate CNS.     

Exocytotic release of vasoactive intestinal polypeptide and serotonin from mucosal nerve fibres and endocrine cells of the intestine of the goldfish (Carassius auratus) and the tilapia (Oreochromis mossambicus): an ultrastructural study

In earlier studies we determined the effect, presence and ultrastructure of vasoactive intestinal polypeptide (VIP) and 5- hydroxytryptamine (5-HT)-containing nerve fibres in the tilapia and goldfish intestinal mucosa. 5-HT-labelled varicosities were found close to the epithelial cells; however, synaptic membrane specializations have never been observed. VIP-like immunoreactive nerve fibres appear to be located less frequently close to the goldfish epithelium, as in the tilapia intestine, in which the distance between the VIP- or 5-HT-labelled varicosities and the epithelial cells was also rather large (more than 2 μm). To establish a possible role of VIP and 5-HT as neurotransmitters involved in the regulation of fish intestinal epithelium, both electron microscopical and immunoelectron microscopical methods were used to visualize the release of 5-HT and VIP from nerve fibres. We found exocytoses from VIP-ergic and serotonergic varicosities in the muscle layers of both fish. Directly underneath the intestinal epithelium of the goldfish, it was demonstrated that 5-HT could be released from scarce varicosities. The release of 5-HT in the tilapia intestinal mucosa could only be observed from endocrine cells     

Exocytotic release of vasoactive intestinal polypeptide and serotonin from mucosal nerve fibres and endocrine cells of the intestine of the goldfish (Carassius auratus) and the tilapia (Oreochromis mossambicus): an ultrastructural study

In earlier studies we determined the effect, presence and ultrastructure of vasoactive intestinal polypeptide (VIP) and 5- hydroxytryptamine (5-HT)-containing nerve fibres in the tilapia and goldfish intestinal mucosa. 5-HT-labelled varicosities were found close to the epithelial cells; however, synaptic membrane specializations have never been observed. VIP-like immunoreactive nerve fibres appear to be located less frequently close to the goldfish epithelium, as in the tilapia intestine, in which the distance between the VIP- or 5-HT-labelled varicosities and the epithelial cells was also rather large (more than 2 μm). To establish a possible role of VIP and 5-HT as neurotransmitters involved in the regulation of fish intestinal epithelium, both electron microscopical and immunoelectron microscopical methods were used to visualize the release of 5-HT and VIP from nerve fibres. We found exocytoses from VIP-ergic and serotonergic varicosities in the muscle layers of both fish. Directly underneath the intestinal epithelium of the goldfish, it was demonstrated that 5-HT could be released from scarce varicosities. The release of 5-HT in the tilapia intestinal mucosa could only be observed from endocrine cells     

Strongyloides ratti: Mast cell and goblet cell responses in the small intestine of infected rats

Kinetics of intestinal mast cells and goblet cells were examined in relation to worm localization at various sites in the small intestine of rats infected with 3000 filariform (stage 3) larvae of Strongyloides ratti. The most marked intestinal mastocytosis was observed on Day 20 at the anterior site of the small intestine where the majority of the worms had concentrated. The number of mast cells in the posterior small intestine increased in parallel with the posterior shift of parasites at the later stage of the infection. In contrast to the intestinal mast cell response, the number of goblet cells was not significantly affected by the infection. These results strongly suggest that intestinal mastocytosis is closely related to the presence of the worms and that mast cells may play an important role for the expulsion of S. ratti.     

尼罗罗非鱼消化道肥大细胞的组化性质

实验采用改良甲苯胺蓝(MTB)、阿利新蓝-沙黄(AB/SO)、甲基绿-派洛宁(MG-P)、天青Ⅱ-伊红-瑞氏混合液和硫堇5种组化染色法,对尼罗罗非鱼(Nile tilapia)消化道组织中的肥大细胞(Mast cell,MC)组化性质进行研究。尼罗罗非鱼的食管、胃及小肠壁内均显示有肥大细胞,在食管和胃的切片标本上肥大细胞主要分布在黏膜固有层和胃腺体之间。在肠道中的肥大细胞主要分布在黏膜固有层和肠上皮下方,少量肥大细胞存在于黏膜下层结缔组织中。细胞呈圆形、椭圆形,也有长梭形的。而且肥大细胞有沿血管分布的特点。5种组化染色结果表明:AB/SO、MTB和MG-P显示的MC效果较好,尤其AB/SO染色效果最好,肥大细胞轮廓清楚,胞质颗粒较清晰;尼罗罗非鱼肥大细胞胞浆颗粒都呈红色,即肥大细胞胞浆主要含肝素,不含组胺。天青Ⅱ-伊红-瑞氏混合液染色效果也很好,但被染的肥大细胞较少;80%乙醇硫堇染色,在尼罗罗非鱼消化道各段组织中均未能鉴定出肥大细胞。尼罗罗非鱼消化道肥大细胞大多分布于浅层的黏膜或血管、腺体周围的结缔组织等易表露于环境抗原的位点。罗非鱼消化道黏膜层结缔组织中的肥大细胞与大多数脊椎动物的肥大细胞一样,具有沿血管分布的特性,说明硬骨鱼的肥大细胞如哺乳动物肥大细胞一样与血管有着密切的关系。       

Mast cells/eosinophilic granule cells of salmonids: staining properties and responses to noxious agents

Observations were made on mast cells/eosinophilic granule cells in swimbladder tissue spreads and sections of gills and intestinal tissues from species of the generaSalmoOncorhynchusSalvelinusCoregonusThymallus. Some individuals had been reared in captivity and others were caught in rivers or lakes, and both apparently healthy fish and fish with persistent inflammation, due to helminths or unknown causative agents, were included in the study. Acute responses to noxious agents were studied in swimbladder tissue spreads after intraperitoneal injections of inactivatedAeromonas salmonicida, compound 48/80 and hydrocortisone. The tissue spreads were fixed in ethanol and stained with thionin. Other tissues were fixed in a solution containing 4% formaldehyde and 5% acetic acid in methanol, and stained with May-Grünwald Giemsa combination dye, haematoxylin and eosin, or Alcian blue. Intestinal tissue histamine was assayed fluorometrically, and vascular responses to histamine and compound 48/80 were studied in perfused gill preparations. The staining properties of salmonid mast cells/eosinophilic granule cells resembled those of mammalian mucosal mast cells and globule leucocytes, with both acidophilic and basophilic components in their granules. May-Grünwald Giemsa staining revealed that in cells found in connective tissues the basophilic character was dominant, whereas the acidophilic character was most marked in those present in epithelia. The granule cells in swimbladder tissue spreads stained metachromatically with alcoholic thionin. Intraperitoneal injections of inactivatedA. salmonicidaproduced acute inflammatory reactions, with degranulation of mast cells/eosinophilic granule cells, in tissues of the swimbladder. Degranulation of the granule cells was also noticed after injection of compound 48/80. Massive degranulation of mast cells/eosinophilic granule cells in the swimbladder wall, followed by an acute inflammatory reaction, was induced by intraperitoneal injections of hydrocortisone. Persistent inflammation, e.g. in tissues infected with helminths, was accompanied by recruitment of mast cells/eosinophilic granule cells. Presence of many or few mast cells/eosinophilic granule cells in tissues of the intestine seemed to have no influence on the content of histamine, which was always low. Compound 48/80 produced increased resistance in the perfused branchial vascular bed, but effects of histamine were slight or completely absent. The responses of mast cells/eosinophilic granule cells of salmonids in acute and persistent inflammation, as revealed in the present investigation, are similar to the known responses of mammalian mast cells. Since their staining properties are also similar, the term ‘mast cell’ should be adequate.     

Degranulation of eosinophilic granule cells induced by capsaicin and substance P in the intestine of the rainbow trout (Oncorhynchus mykiss Walbaum)

Summary Adult rainbow trout (Oncorhynchus mykiss) were injected intraperitoneally with capsaicin, substance P, serotonin, or a control of saline vehicle or bovine serum albumin (0.5 g/g body weight). Fish were sacrificed 30 min and 1,2 and 4 h post-injection, the gut was dissected out, and a small section of the upper intestine was processed for electron microscopy. A significant proportion of eosinophilic granule cells (EGCs) of the intestine were in close association with non-myelinated neuronal bundles in all fish (4 fish per treatment and time period), but there was no significant difference between treatment or time, suggesting that the association was unaffected by these factors. Close examination of EGC ultrastructure showed that fish treated with capsaicin and substance P exhibited limited degranulation of the EGCs in the stratum compactum and extensive crinophagic-like degranulation in the lamina propria. Cells of the lamina propria contained characteristic multivesicular-like bodies. The degranulation was reminiscent of both mast cell degranulation and endocrine cell crinophagy. EGCs of fish treated with serotonin or a control were unaffected, suggesting that the serotoninergic neurons, believed to be involved in gut motility, were not responsible for degranulation. It is apparent that EGCs of the trout intestine may be under nervous control, as has been demonstrated previously for mammalian mast cells.     

禽沙门菌诱导宿主免疫应答的特性

沙门菌病(Salmonellosis)是全世界最普遍的食源性疾病之一,不仅对养殖业造成经济损失,还对人类安全构成威胁。禽沙门菌感染肠道后,可诱导肠上皮细胞表达多种TLRs和炎症反应的发生,在分泌的趋化因子作用下免疫效应细胞迁移到感染部位。细菌通过肠上皮细胞屏障后被巨噬细胞或树突状细胞吞噬,其中巨噬细胞是沙门菌的主要定殖场所。天然免疫系统将抗原递呈给淋巴细胞后,机体能够在2–3周内通过以Th1为主的免疫应答清除在肠道和深层组织中的沙门菌。而宿主特异性血清型鸡白痢沙门菌从肠道侵入后,在肝脾和其他器官中定殖,进而引发全身感染。早期感染阶段不会引起肠道炎症反应,主要诱导以Th2为主的免疫应答,而Th1型应答相对较弱,有利于鸡白痢沙门菌在机体内的持续存在和感染。本文围绕禽沙门菌的致病机理和免疫应答特性进行阐述,尤其对鸡白痢沙门菌免疫逃逸和持续载菌的特性进行深入分析,为禽沙门菌病的防控提供新靶标和新见解。     

Depletion of head kidney neutrophils and cells with basophilic granules during peritoneal inflammation in the goldfish, Carassius auratus

The head kidney morphology of the goldfish with the experimental peritoneal inflammation (on day 2 after i.p. injection with sterile 3% Thioglycollate) is compared with that in the control fish (i.e. on day 2 after i.p. injection with a strile physiological saline PBS) with special emphasis on identification of granulocytes on semithin and consecutive ultrathin sections. The most striking feature of head kidneys of goldfish in the course of peritoneal inflammation is a severe depletion of mature neutrophils and cells with basophilic granules (basophils/mast cells). These observations suggest the involvement of the head kidney, the main hematopoietic organ of teleosts, in the inflammatory process.     

Effect of alimentary deprivation on quantitative and time characteristics of nutrition of the goldfish in norm and under action of adrenalin

Effects of the intraperitoneally administered adrenalin (0.14 mg/kg) on volume and time characteristics (duration) of the individual, grouped, and summarized nutrition was studied in goldfish Carassius auratus (L.) previously kept on normal and restricted (for 10 and 15 days) ration. The fish on restricted ration demonstrated the two-phase alimentary response to the adrenalin injection similar with that in normally feeding fish: a decrease in volume and in the grouped and summarized duration of nutrition in the first phase of the hormone action and an increase-in the second phase; however, the value of the effect in the first phase was lower, while in the second phase-higher than in the fish that obtained before the experiment the sufficient amount of food. Duration of individual nutrition increased in all fish groups, including the control one, the maximal increase being observed in goldfish submitted to alimentary deprivation. The content of glycogen in hepatopancreas of the normally fed goldfish by the end of experiment exceeded that in fish restricted in nutrition 4.5–6 times. The obtained results confirm the suggestion that a decrease of level of reserved substances in the body leads to reduction of susceptibility of fish to the stress hormoneadrenalin and, hence, to a change of the value of its effects on the goldfish food-procuring activity, with preservation of dynamics of parameters.     

肥大细胞在动脉粥样硬化形成中的作用

动脉粥样硬化,是冠心病的病理基础,被认为是一种慢性炎症性疾病,涉及如巨噬细胞和T淋巴细胞等许多炎性细胞。肥大细胞是一种重要的免疫细胞,其功能主要是在超敏反应方面的作用。有病理学研究表明:肥大细胞在动脉粥样硬化斑块周围表达增加,这表明肥大细胞可能与疾病的进展有关。最近的研究表明,肥大细胞在动脉粥样硬化中确实起着重要的作用。本文通过总结肥大细胞在动脉粥样硬化形成中的作用,为在疾病进程中,通过调节肥大细胞功能来改善动脉粥样硬化的这种治疗方式的可能性提供依据。     

The role of interleukin-33 in pathogenesis of bronchial asthma. New experimental data

Interleukin-33 (IL-33) belongs to the IL-1 cytokine family and plays an important role in modulating immune system by inducing Th2 immune response via the ST2 membrane receptor. Epithelial cells are the major producers of IL-33. However, IL-33 is also secreted by other cells, e.g., bone marrow cells, dendritic cells, macrophages, and mast cells. IL-33 targets a broad range of cell types bearing the ST2 surface receptor. Many ST2-positive cells, such as Th2 cells, mast cells, basophils, and eosinophils, are involved in the development of allergic bronchial asthma (BA). This suggests that IL-33 directly participates in BA pathogenesis. Currently, the role of IL-33 in pathogenesis of inflammatory disorders, including BA, has been extensively investigated using clinical samples collected from patients, as well as asthma animal models. In particular, numerous studies on blocking IL-33 and its receptor by monoclonal antibodies in asthma mouse model have been performed over the last several years; IL-33-and ST2-deficient transgenic mice have also been generated. In this review, we summarized and analyzed the data on the role of IL-33 in BA pathogenesis and the prospects for creating new treatments for BA.     

The establishment of an intestinal microflora in developing goldfish (Carassius auratus) of culture ponds

The bacterial flora in the intestinal tract of goldfish (Carassius auratus) was investigated at different stages of fish development. The floras of the diets and the water and sediment of a culture pond were also analyzed. The total counts in the intestine ranged from 2.2 × 10⁶–2.1 × 10⁸ cells g^–1 wet weight.Aeromonas hydrophila, A. punctata, Pseudomonas, Bacteroidaceae andClostridium species were the common components in the intestinal tract of goldfish from larvae to adult stage.Bacteroides type A appeared at 44 days with a density of 10³ cells g^–1 and then predominated with densities of 10⁵–10⁷ cells g^–1. The intestinal microflora of goldfish become relatively stable after 67 days of hatching. These observations suggest that the intestinal microflora of adult goldfish becomes established approximately 2 months after hatching.     

肥大细胞在动脉粥样硬化形成中的作用

动脉粥样硬化,是冠心病的病理基础,被认为是一种慢性炎症性疾病,涉及如巨噬细胞和T淋巴细胞等许多炎性细胞。肥大细胞是一种重要的免疫细胞,其功能主要是在超敏反应方面的作用。有病理学研究表明：肥大细胞在动脉粥样硬化斑块周围表达增加,这表明肥大细胞可能与疾病的进展有关。最近的研究表明,肥大细胞在动脉粥样硬化中确实起着重要的作用。本文通过总结肥大细胞在动脉粥样硬化形成中的作用,为在疾病进程中,通过调节肥大细胞功能来改善动脉粥样硬化的这种治疗方式的可能性提供依据。