Expression of conserved mucin domains by epithelial tissues in various mammalian species

Mucins are related to infectious and non-infectious diseases in Veterinary and Human Medicine. MUC1 mucin is a transmembrane glycoprotein expressed on the apical surface of human epithelia while MUC5AC is the predominant secreted mucin expressed in human gastric epithelium and goblet cells of lung and eyes. MUC5AC C-terminus cysteine rich regions and the cytoplasmic tail of MUC1 domains are conserved among several mammalian species. Objective: to compare the expression of MUC1 and MUC5AC mucins in mammalian epithelia. CT33 anti-MUC1 cytoplasmic tail (MUC1CT) polyclonal antibody and 45M1 anti-MUC5AC monoclonal antibody were employed. By immunohistochemistry, MUC1CT was expressed in most tissues while MUC5AC was restricted to gastric surface epithelium and goblet cells from trachea and lung. By western blot, MUC1CT showed a band at approximately 35 kDa in most tissues; MUC5AC revealed bands at >180 kDa in stomach and lung secretions from rat, cat, pig and cow. When rat MUC5AC was immunoprecipitated, a band at about 180 kDa was obtained.     

Goblet Cell Types in Intestine of Tiger Barb and Black Tetra (Cyprinidae,Characidae: Teleostei)

Histochemical properties of goblet cells in intestine of a stomach‐less teleost, tiger barb (Puntius tetrazona), and a stomach‐containing teleost, black tetra (Gymnocorymbus ternetzi), are described and compared. The intestine goblet cells were mostly wide in both species, but in tiger barb, some of them were markedly thinner. In black tetra, all the intestine goblet cells displayed magenta colour after PAS, whereas in the tiger barb, only the thinner goblet cells displayed such affinity. The latter cell type was coloured strongly magenta when the tissue was treated with alcian blue (pH 2.5) followed by PAS, whereas the wide goblet cells in tiger barb and all goblet cells in black tetra displayed mainly a blue colour after such treatment. Further, the goblet cells in both species were coloured cleanly blue after high iron diamine followed by alcian blue (pH 2.5). The intestine goblet cells in both species displayed a moderate affinity to WGA and concanavalin A lectins and no affinity to DBA. Most of the goblet cells displayed no affinity to PNA, but some of them in the tiger barb displayed a moderate or strong affinity to this lectin. The affinity to WGA was somewhat strengthened after pre‐treatment with neuraminidase. These results suggest that tiger barb contains two types or variants of intestinal goblet cells: high numbers of wide cells filled by acidic, non‐sulphated mucin and some thinner cells filled by neutral mucin. The intestine goblet cells in black tetra were filled by variable amounts of neutral and acidic mucin, but the total number of such cells is much less than in tiger barb. The present lectin and neuraminidase results suggest that the intestinal mucins in both species contain significant amounts of N‐acetylglucosamine, sialic acid and glucose/mannose, but seem to lack N‐acetylgalactosamine. However, some of these cells in tiger barb contain moderate to large amounts of galactose. Together, these results suggest significant species‐specific features of the intestine goblet cells and mucin types in tiger barb and black tetra. In conclusion, the present results suggest that the diet and feeding habits in stomach‐less teleosts compared with stomach‐containing teleosts, greatly influence the number of intestine goblet cells and type of mucin in these cells.     

An immunohistochemical study of S-100 protein in the intestinal tract of Chinese soft-shelled turtle, Pelodiscus sinensis

The present work describes the distribution of S-100 protein in the intestinal tract of a Chinese soft-shelled turtle specimen (Pelodiscus sinensis). S-100 protein positive cells were located in the intestinal tract, from the proximal small to distal large intestine. S-100 protein positive dendritic cells had irregular shape and were positive in both cytoplasm and nucleus. Most of them were located both lamina propria and submucosa in the small intestine, while few were found in the large intestine. S-100 protein, C-kit positive ICCs and Silver staining glial cells were predominantly observed in three locations: (1) in the interspace between the submucosa and circular muscle layer; (2) in the circular muscle layer; and (3) between the circular and longitudinal muscle layers of the intestine. Fewer were found in the large intestinal lamina propria and submucosa. Three types of positive cells (S-100 protein positive cells, C-kit positive ICCs and Silver staining glial cells) with 1–2 long or 2–3 short processes were distributed as lace-like or surrounding blood vessels in the different locations mentioned above. In the lamina propria, all the positive cells with irregular processes were connected with each other and formed a network. In the submucosa, all the positive cells were found surrounding the blood vessels.     

Characterization of mucin gene expression and goblet cell proportion in inflammatory colorectal polyps in miniature dachshunds

Hyperplastic goblet cells and abundant mucus are significant characteristics of inflammatory colorectal polyps (ICRPs) in miniature dachshunds. In this study, selected mucin gene expressions and goblet cell proportions were evaluated in miniature dachshunds with ICRPs and in healthy dogs. Mucin 2 (MUC2) gene expression was not significantly different among the groups, whereas mucin 5AC (MUC5AC) gene expression was significantly higher in the polypoid lesions than in healthy colonic mucosa. Although the percentage of goblet cells in the upper crypt regions did not significantly differ between the groups, that in the lower crypt regions was significantly decreased in polypoid lesions. In conclusion, increased MUC5AC gene expression and goblet cell proportion changes may be associated with the pathogenesis of ICRPs.     

Enteric campylobacter infection in gnotobiotic calves and lambs

Gnotobiotic calves and lambs were infected orally with Campylobacter jejuni, C coli or C hyointestinalis to assess pathogenicity. All animals were successfully colonised and excreted mucoid faeces but showed no other clinical signs. Campylobacters colonised the large intestine better than the small intestine, in which bacterial numbers decreased with time after infection. Campylobacters were found occasionally in the lumen of crypts in close proximity to epithelial cells and included in a mucus-like material. Lesions were mostly in the large intestine in calves whereas in lambs they were present in the ileum. In animals inoculated with C jejuni or C coli scattered crypt abscesses, focal inflammatory infiltrates in the lamina propria and goblet cell discharge were found. In lambs inoculated with C hyointestinalis only minor changes were found in the small intestine. Serum antibody response was either absent or present at a low level only from the 19th day after infection.     

Gut-associated lymphoid tissue in the large intestine of calves. II. Electron microscopy

Scanning electron microscopy of lymphoid tissue in the large intestine of three germfree calves (age 3, 6, and 7 days) revealed two different units: propria nodules and lymphoglandular complexes (LGC). Propria nodules had lymphoid tissue predominantly in lamina propria and were covered by distinct follicle-associated epithelium which lacked goblet cells; nodules were surrounded by wide crypts, which were also lined by follicle-associated epithelium towards the luminal side. Lymphoglandular complexes had lymphoid follicles in the tunica submucosa; epithelial diverticulae extended through the muscularis mucosae branching into the lymphoid nodule. In centers of lymphoglandular complexes, protrusions of lymphoid tissue were covered with distinct follicle-associated epithelium. By transmission electron microscopy cells compatible with M cells in the small intestine of calves and cells with characteristics of both enteroabsorptive and M cells were found. Follicle-associated epithelium of propria nodules and lymphoglandular complexes differed only in the relative frequency of cell types.     

The Prenatal Development and Histochemistry of the Ileal Mucins in the Bovine Fetuses

Few studies exist regarding the distribution of intestinal mucins in fetuses of mammalians such as cattle and sheep. In this study, we aimed to describe the changes in the mucin production by ileal epithelium of bovine fetuses during their prenatal development. The goblet cells showed heterogeneity in mucins and the apical cytoplasm of the enterocytes demonstrated Periodic acid Schiff‐positive reaction which declined gradually towards the birth. Moreover, the number of the goblet cells containing acidic and mixed mucins augmented, whereas those containing neutral mucins decreased with advancing gestational age. After sixth month of gestation, with the initiation of the ileal Peyer patches and follicle‐associated epithelium development, a gradual increase in the number of goblet cells containing sulfomucins was also noticed towards the birth. The presence of different mucins in the ileum of bovine fetuses throughout prenatal development might play a role in the protection of the intestinal mucosa against urinary waste products in swallowed amniotic fluid and bile. Furthermore, mucins can also contribute for the formation of meconium in intra‐uterine life and building of strong intestinal barrier with predominating sulfomucins, protecting the intestine against potential pathogens and digestive enzymes after birth.     

Expression of secreted and membrane-bound mucins in the airways of piglets experimentally infected with Mycoplasma hyopneumoniae

Immunohistochemistry was used to demonstrate secreted mucins MUC2, MUC5AC and MUC5B and membrane-bound mucin MUC4 in the pulmonary bronchioles of piglets experimentally infected with Mycoplasma hyopneumoniae. Conventional status, Landrace-Duroc cross-bred piglets, 13 days of age, were randomised to two groups. One group (n=20) was infected by the intra-tracheal route with the SNU98703 strain of M. hyopneumoniae, and a group of 12 animals acted as uninfected controls. Five infected and three uninfected piglets were euthanased on the day of infection and at 7, 21, and 35 days post-inoculation (PI). Membrane-bound MUC4 and secreted MUC5AC were the predominant mucins produced in the bronchioles of the piglets in response to M. hyopneumoniae infection, but by day 35 PI, all labelled mucins had returned to pre-infection levels, contemporaneous with reduced pulmonary lesion scores. The increased mucin production may result from direct stimulation of the epithelium by mycoplasmal infection, or may arise indirectly following M. hyopneumoniae-induced ciliostasis.     

贝氏莫尼茨绦虫感染绵羊对小肠局部黏膜淋巴组织的影响

为了研究贝氏莫尼茨绦虫自然感染绵羊对小肠黏膜免疫组织的影响,分别从宏观、微观及亚微观水平对自然感染贝氏莫尼茨绦虫的成年绵羊(感染组)肠道进行了细致地观察,并与正常成年绵羊(正常组)进行了比较.结果显示,感染组肠道所见虫体平均长度为1.5m,头节主要吸附在空肠淋巴集结分布丰富的部位,一般寄生数量为1～2条.眼观,虫体寄生部位黏膜增厚,表面有大量灰白色黏液附着,其间可见点状出血.镜下,局部黏膜上皮脱落,而在完整的黏膜上皮处,其上皮细胞、上皮内淋巴细胞、杯状细胞的数量都明显增多;固有层内毛细血管充血,淋巴细胞、浆细胞、弥散淋巴组织以及肠腺杯状细胞均有不同程度的增生,头节寄生处部分肠腺坏死;黏膜下层淋巴小结、淋巴集结显著增生,部分增生凸入固有层形成新的圆顶区;固有层与黏膜下层以及黏膜肌层可见大量嗜酸性粒细胞浸润.扫描电镜下,感染组肠黏膜上皮脱落;贝氏莫尼茨绦虫头节呈椭球状,有4个吸盘,无顶突,小沟,表面覆盖一层致密的微绒毛.研究结果表明,肠黏膜增厚,主要是局部黏膜免疫相关细胞在寄生虫虫体表面覆盖的微绒毛的不断刺激下,机体抗感染自身组织增生所致.成年绵羊对抗贝氏莫尼茨绦虫的感染可能是通过黏膜免疫相关组织增生来加强局部免疫力而实现的.     

Histology and Mucin Histochemistry of The Digestive Tract of Yellow Catfish, Pelteobagrus fulvidraco

The histology and characteristics of mucins secreted by epithelial mucous cells of the digestive tract in yellow catfish, Pelteobagrus fulvidraco were investigated using light microscope and transmission electron microscope. The digestive tract was divided into a pharynx, oesophagus, U-shaped stomach (with a cardiac, fundic and pyloric part) and intestine, composed of anterior intestine, middle intestine and posterior intestine, which consisted of a mucosa (epithelial layer), lamina propria-submucosa, muscularis and serosa. A large number of isolated longitudinal striated muscular bundles were present in the lamina propria-submucosa of pharynx. Goblet cells were observed throughout the digestive tract, except in the stomach. In the cardiac and fundic stomach, a plenty of gastric glands were observed, whereas they were absent in the pyloric part. Numerous mitochondria and endoplasmic reticulum were observed in the columnar epithelial cells of the intestine, especially of the anterior part. The epithelial mucous cells contained neutral or other two mixtures of acid and neutral mucins, the first being the most common. The neutral mucin was the only type of mucins in the stomach, anterior intestine and middle intestine. The results of this study will be helpful for understanding the digestive physiology and diagnosing some gastrointestinal diseases in yellow catfish.     

丁酸钠对肉鸡小肠黏膜免疫相关细胞的影响

本试验通过日粮中添加丁酸钠研究其对肉鸡小肠黏膜免疫相关细胞的影响.将32只1日龄健康AA肉鸡随机分为4个处理组:A组(对照组:基础日粮)、B组(丁酸钠组:基础日粮 500mg/kg丁酸钠)、C组(丁酸钠 芽孢杆菌组:基础日粮 100 mg/kg丁酸钠 200 mg/kg芽孢杆菌)和D组(丁酸钠 酶制剂组:基础日粮 100 mg/kg丁酸钠 500 mg/kg酶制剂).试验7周后,采用组织学技术研究肉鸡小肠黏膜免疫相关细胞的分布变化.结果显示:(1)与对照组比较,小肠肥大细胞、上皮内淋巴细胞及杯状细胞的形态无明显变化,肥大细胞多存在于肠腺周围和肠绒毛固有层,(2)各试验组的小肠肥大细胞、上皮内淋巴细胞及杯状细胞数均显著高于对照组(P<0.05);(3)各试验组和对照组小肠肥大细胞和上皮内淋巴细胞数均从前向后逐渐减少,十二指肠的肥大细胞最多,空肠的次之,回肠的较少;杯状细胞数则与之相反.由此可知,3种添加剂可不同程度地改善肉鸡小肠黏膜结构,且将丁酸钠分别与芽孢杆菌和酶制剂混合饲喂,效果在一定程度上好于单独饲喂丁酸钠.     

Micromorphological studies on the small intestine and caeca in wild and captive willow grouse (Lagopus lagopus lagopus)

The present study describes the micromorphology of the small intestine and caeca of wild and captive willow grouse. The micromorphology of the small intestine was similar in wild and captive birds, while typical differences were apparent in the caeca, Wild grouse had ciliated epithelium without goblet cells in the neck part of the caeca, captive birds had strongly atrophied cilia and a high number of goblet cells. The epithelium of the body part of caeca of wild birds lodged a great number of spiral-shaped microorganisms and amoebae, which were absent in captive birds. Both the caecal villi and the longitudinal folds were much larger in wild than captive grouse. In the captive grouse the caecal lamina propria was heavily infiltrated with mono-nucleated cells and very often also with polymorphonucleated heterophilic leucocytes.Since only first generation captives were used in this study, the differences in gut morphology of captive and wild grouse must be due to different food and environmental conditions and not to genetic selection.     

Goblet Cells and Mucus Types in the Digestive Intestine and Respiratory Intestine in Bronze Corydoras (Callichthyidae: Teleostei)

The structure and histochemical properties of the intestine in bronze corydoras (Corydoras aeneus), a stomach‐containing teleost, are described, with emphasis on goblet cells and mucin types. The proximal intestine displayed a normal structure for teleosts, whereas the distal intestine was wide, translucent, thin‐walled, richly vascularized and constantly filled with air, suggesting an important respiratory role. Goblet cells were common throughout the entire intestine and displayed a variable, but mainly faint metachromatic colour after toluidine blue. They were moderately coloured by alcian blue at both pH 2.5 and 0.2 and displayed no colour after periodic acid followed by Schiff's solution (PAS), but a distinct purple‐brown colour after high iron diamine followed by alcian blue (pH 2.5). Together, these results suggest that the mucin in the intestine goblet cells consists mainly of sulphated proteoglycans. Further, the results from the present lectin and neuraminidase tests suggest that these mucins contain much N‐acetylglucoseamines and some N‐acetylgalactosamines and sialic acid, but seem to lack glucose and mannose. They also contain some galactose‐N‐acetylgalactosamines sequences, normally hidden by sialic acid. The distinct brush border and mucus layer on the epithelial cells in the respiratory intestine may indicate some digestive roles, such as absorption of water, ions and simple carbohydrates. As sulphated proteoglycans are tough and attract much water, this mucus may play important roles in the protection against mechanical and chemical damages and in the defence against micro‐organisms throughout the entire intestine, but in the respiratory intestine it may impede significantly the oxygen uptake. However, as this part of the intestine usually contains no digesta, but is completely filled with air, frequently renewed by dry air from the atmosphere, and the main function of the mucus may be to protect the respiratory epithelium against a destroying and dangerous desiccation.     

Expression of secreted mucins (MUC2, MUC5AC, MUC5B, and MUC6) and membrane-bound mucin (MUC4) in the lungs of pigs experimentally infected with Actinobacillus pleuropneumoniae

The expression patterns of different secreted (MUC2, MUC5AC, MUC5B, and MUC6) and membrane-bound (MUC4) mucins were determined immunohistochemically in the lungs of pigs experimentally infected with Actinobacillus pleuropneumoniae. Forty-seven-week-old colostrum-deprived pigs were randomly allocated to infected (n=20) or control groups (n=20). Five infected and uninfected pigs were euthanized at 0, 6, 12, and 48 h post-inoculation (hpi). In the infected pigs, the expression of both types of mucins, which were invariably observed, was associated with bronchiolar and respiratory bronchiolar lesions. Strong positive mucin signals were seen on the surface of bronchiolar and respiratory bronchiolar epithelium with neutrophil infiltration. The mean mucin-positive area peaked at 6 hpi and decreased significantly to control levels by 48 hpi on the surface of the bronchiolar and respiratory bronchiolar epithelium. Further studies are needed to establish the functional relationship between mucin expression and the host defense mechanism against A. pleuropneumoniae in the lungs of infected pigs.     

Effect of added dietary threonine on growth performance,health, immunity and gastrointestinal function of weaning pigs with differing genetic susceptibility to Escherichia coli infection and challenged with E. coli K88ac

Threonine (Thr) is important for mucin and immunoglobulin production. We studied the effect of added dietary Thr on growth performance, health, immunity and gastrointestinal function of weaning pigs with differing genetic susceptibility to E. coli K88ac (ETEC) infection and challenged with ETEC. Forty‐eight 24‐day‐old weaned pigs were divided into two groups by their ETEC susceptibility using mucin 4 (MUC4) gene as a marker (2 MUC4^?/?, not‐susceptible, and 2 MUC4^+/+, susceptible, pigs per litter). Within genotype, pigs were fed two different diets: 8.5 (LThr) or 9.0 (HThr) g Thr/kg. Pigs were orally challenged on day 7 after weaning and slaughtered on day 12 or 13 after weaning. Before ETEC challenge, HThr pigs ate more (p < 0.05). The diet did not affect post‐challenge growth, but HThr tended to increase post‐challenge feed efficiency (p = 0.087) and overall growth (p = 0.087) and feed efficiency (p = 0.055). Before challenge, HThr pigs excreted less E. coli (p < 0.05), while after challenge, diet did not affect the number of days with diarrhoea and ETEC excretion. MUC4^+/+ pigs responded to the challenge with more diarrhoea, ETEC excretion and anti‐K88 IgA in blood and jejunal secretion (p < 0.001). HThr pigs had a higher increase of anti‐K88 IgA values in jejunal secretion (p = 0.089) and in blood (p = 0.089, in MUC4^+/+ pigs only). Thr did not affect total IgA and IgM values, morphometry of jejunum, goblet cells count in colon, total mucin from jejunum and colon, but varied jejunal goblet cells counts (p < 0.05). In the first two post‐weaning weeks, 8.5 g Thr/kg diet may be not sufficient to optimize initial feed intake, overall feed efficiency and intestinal IgA secretion and to control the gut microbiota in the first post‐weaning week, irrespective of the pig genetic susceptibility to ETEC infection.     

野生黄鼬消化管组织结构的观察

通过组织学观察,探讨野生黄鼬消化管的组织结构特点.将8只野生黄鼬经乙醚麻醉后处死,解剖取食管、胃、小肠、大肠,制作石蜡切片,观察其组织结构.结果显示野生黄鼬食管的黏膜为复层扁平上皮,食管腺发达,肌层以骨骼肌为主.胃贲门部有发达的皱襞和贲门腺;胃底腺有大量的主细胞和壁细胞;胃大弯部的腺体以壁细胞为主,仅有少量主细胞;胃幽门部有发达的幽门腺和大量壁细胞.十二指肠黏膜层有小肠腺,内有潘氏细胞存在,黏膜下层含有十二指肠腺;空肠可见孤立淋巴小结、弥散淋巴组织及集合淋巴小结.结肠无皱襞和肠绒毛,大肠腺排列紧密,其中杯状细胞特别多;直肠固有膜内有发达的大肠腺.所以野生黄鼬消化管的特点是胃各部胃腺发达,壁细胞特别多.     

Structure of the Oesophagus and Morphometric, Histochemical–Immunohistochemical Profiles of the Oesophageal Gland During the Post-hatching Period of Japanese Quails (Coturnix coturnix japonica)

In the oesophagus, mucins, which originate from oesophageal submucosal glands, play an important role in the mucosal protection as a pre-epithelial barrier. In this study, the structure of cervical and thoracic parts of oesophagus of Japanese quail during the post-hatching period was compared, and the contents of carbohydrate and gastric mucin MUC5AC of the oesophageal glands in these parts were analysed at the light microscope levels by applying conventional histochemical and immunohistochemical methods. The oesophageal glands were present at hatching, located in the laminae propria. The numbers of glands were different in the cervical and thoracic parts, but the differences were found to be insignificant. The thoracic part has the oesophageal tonsils which are associated with the glands. Oesophageal tonsil was formed from day 5 after hatching. In quail of all ages, the secretory epithelium of glands contained neutral sialomucins and weakly sulphomucins. The cells in the neck region of secretory units contained sialomucins, while the cells of excretory ducts had strongly sulphomucins. Sialomucin containing cells in the secretory units increased with the advance of age and glandular development. But, in the secretory units, the sulphomucin content of glands was more in the thoracic part. The secretory epithelium of tonsil-associated glands contained mostly sulphomucins and a little sialomucin. From the hatching, MUC5AC mucin was detected in the cells of excretory ducts. Although the lymphoepithelium of the tonsil units exhibited negative reactions to all histochemical methods, it showed positive reaction to MUC5AC mucin antibody. In conclusion, the cervical and thoracic parts may be functionally different and the thoracic part of oesophagus was transformed into an immunological organ following day 5 after hatching.     

An immunohistochemical study of S-100 protein in the intestinal tract of Chinese soft-shelled turtle,Pelodiscus sinensis

The present work describes the distribution of S-100 protein in the intestinal tract of a Chinese soft-shelled turtle specimen (Pelodiscus sinensis). S-100 protein positive cells were located in the intestinal tract, from the proximal small to distal large intestine. S-100 protein positive dendritic cells had irregular shape and were positive in both cytoplasm and nucleus. Most of them were located both lamina propria and submucosa in the small intestine, while few were found in the large intestine. S-100 protein, C-kit positive ICCs and Silver staining glial cells were predominantly observed in three locations: (1) in the interspace between the submucosa and circular muscle layer; (2) in the circular muscle layer; and (3) between the circular and longitudinal muscle layers of the intestine. Fewer were found in the large intestinal lamina propria and submucosa. Three types of positive cells (S-100 protein positive cells, C-kit positive ICCs and Silver staining glial cells) with 1–2 long or 2–3 short processes were distributed as lace-like or surrounding blood vessels in the different locations mentioned above. In the lamina propria, all the positive cells with irregular processes were connected with each other and formed a network. In the submucosa, all the positive cells were found surrounding the blood vessels.     

Mycotoxicosis caused by aerosolized T-2 toxin administered to female mice

Thymus, spleen, adrenal glands, and small intestine of female mice exposed to aerosolized T-2 mycotoxin were examined at postexposure hours (PEH) 0.25, 1, 2, 4, 6, 9, 12, and 24. Lymphocyte necrosis was observed at PEH 1 in the thymus, spleen, and lamina propria and Peyer patches of the small intestine. Necrosis of small intestinal crypt epithelial cells was observed at PEH 2, and necrosis of parenchymal cells and increased number of neutrophils were seen in sinusoids of the adrenal cortex at PEH 4. These results indicated that the earliest microscopic evidence of T-2 mycotoxicosis after aerosol exposure was necrosis of lymphocytes in the thymus, spleen, and lamina propria and Peyer patches of the small intestine.