Immunohistochemical localization of galectin-3 in the granulomatous lesions of paratuberculosis-infected bovine intestine

The presence of galectin-3 was immunohistochemically quantified in bovine intestines infected with paratuberculosis (Johne''s disease) to determine whether galectin-3 was involved in the formation of granulation tissue associated with the disease. Mycobacterium avium subsp. paratuberculosis infection was histochemically confirmed using Ziehl-Neelsen staining and molecularly diagnosed through rpoB DNA sequencing. Galectin-3 was detected in the majority of inflammatory cells, possibly macrophages, in the granulomatous lesions within affected tissues, including the ileum. These findings suggest that galectin-3 is associated with the formation of chronic granulation tissues in bovine paratuberculosis, probably through cell adhesion and anti-apoptosis mechanisms.     

Immunohistochemical localization of IgG1 and IgG2 in prepartum and lactating bovine mammary tissue

The differential distributions of IgG1 and IgG2 were determined in prepartum and lactating bovine mammary tissue by indirect immunofluorescence. IgG1 was found predominately within the alveolar epithelial cells and lumens of prepartum tissue whereas IgG2 was largely confined to the stromal area surrounding the alveoli. Both IgG subclasses were confined predominately to the stroma in lactating tissue. Few IgG containing stromal cells were readily distinguished in any of the mammary tissue used in this study.     

Immunohistochemical localization of 3beta-hydroxysteroid dehydrogenase in the granulosa and theca interna layers of bovine cystic follicles

The aim of the present study was to determine whether the alteration of population of cells containing 3beta-hydroxysteroid dehydrogenase (3beta-HSD) is responsible for the formation of cystic follicles. Paraffin sections of healthy (2 to 5 mm in diameter), atretic (2 to 5 mm) and cystic follicles (more than 25 mm) were immunohistochemically stained with rabbit polyclonal antibody to bovine 3beta-HSD. The 3beta-HSD-positive cells were counted in 4 different regions of the follicles from the apical to the basal side. The frequencies of 3beta-HSD-positive granulosa cells in cystic follicles were significantly higher than those in the healthy follicles (P<0.05), although the number of 3beta-HSD-positive granulosa cells in the cystic follicle were fewer than half the cells (30 to 40%) and was much smaller than that in preovulatory follicles (Conley et al., 1995). The frequencies of 3beta-HSD-positive cells were higher in the granulosa layer and lower in the theca interna layer of the cystic follicles than the atretic follicles. These results suggest that the differentiation of granulosa cells to express 3beta-HSD might be insufficient in cystic follicles and accordingly they fail to ovulate. The differences of frequencies of 3beta-HSD-positive cells in the granulosa and theca interna layers between cystic and atretic follicles may be one of the reasons why regression is delayed in cystic follicles.     

Immunohistochemical localization of haptoglobin in porcine lungs

The extravasation of erythrocytes into the lower respiratory tract occurs in numerous lung injuries and may lead to oxidative damages in lung tissues. Haptoglobin (Hp), the major haemoglobin-binding protein, is known to reduce lung injury associated with exposure to blood in mice. In pigs, Hp is a major acute phase protein and its serum concentrations are elevated in various infections of the respiratory tract. However, information on the porcine Hp response towards inflammatory stimuli is restricted to blood. We herein investigated the presence of Hp in lung tissues from pigs with acute and chronic bronchopneumonia via immunohistochemistry. Hp was localized in airway epithelial cells and immigrated leucocytes whereas in alveolar epithelial cells there was no distinct signal. Unaltered lungs showed less Hp-positive cells compared with lungs from pigs with acute or chronic bronchopneumonia.     

Immunohistochemical localization of Pasteurella haemolytica A1-derived endotoxin, leukotoxin, and capsular polysaccharide in experimental bovine Pasteurella pneumonia

Six, 5- to 10-week-old male Holstein calves were inoculated intratracheally with 5 x 10(9) logarithmic growth phase Pasteurella haemolytica biotype A serotype 1 (A1). Immunohistochemical techniques in conjunction with the use of monoclonal antibodies directed against P. haemolytica A1-derived lipopolysaccharide (LPS), capsular polysaccharide, and a polyclonal rabbit anti-leukotoxin antibody were used to localize their respective antigens in tissue sections of pneumonic lung at the light and electron microscopic levels. We found the following: 1) LPS, capsular polysaccharide, and leukotoxin were released into the inflammatory exudate; 2) LPS was found within the cytoplasm of neutrophils (located in the alveolus and alveolar wall), alveolar macrophages, endothelial cells, pulmonary intravascular macrophages, and on epithelial cell surfaces; 3) capsular polysaccharide was found in the alveolus and alveolar macrophages but not in cells of the alveolar wall; and 4) leukotoxin was associated with cell membranes of degenerating inflammatory cells located in the alveolus. This is the first study that demonstrates the presence of leukotoxin in the pulmonary inflammatory lesions caused by P. haemolytica A1 and implicates endotoxin as an important factor in the genesis of the pulmonary lesions.     

GnRH受体在山羊结状神经节的分布

取山羊结状神经节5对,采用免疫组织化学SP法观察促性腺激素释放激素(Gonadotropin releasing hormon,GnRH)受体在结状神经节的分布特点。结果显示,GnRH受体在结状神经节上广泛分布,神经元、卫星细胞、神经纤维、血管内皮细胞均有不同程度的免疫阳性染色。在神经元胞体中,细胞膜和细胞质有GnRH受体强阳性产物分布,核不着色;卫星细胞、血管内皮细胞也有强阳性产物分布;神经纤维存在弱阳性产物分布。图像分析结果表明,神经元中GnRH受体的相对表达量与其他非神经结构相比差异性显著(P<0.05)。结果表明,山羊结状神经节中的GnRH受体主要存在于神经元中,山羊结状神经节中的内脏感觉传入神经元对GnRH具有反应性,提示GnRH结状神经节内脏感觉传入神经元的GnRH受体可能作为GnRH对内脏器官的内分泌调节和自主神经对内脏器官的神经调节这2种途径协调作用的节点。     

GnRH受体在山羊结状神经节的分布

取山羊结状神经节5对,采用免疫组织化学SP法观察促性腺激素释放激素（Gonadotropin releasing hormon,GnRH）受体在结状神经节的分布特点。结果显示,GnRH受体在结状神经节上广泛分布,神经元、卫星细胞、神经纤维、血管肉皮细胞均有不同程度的免疫阳性染色。在神经元胞体中,细胞膜和细胞质有GnRH受体强阳性产物分布,核不着色;卫星细胞、血管内皮细胞也有强阳性产物分布;神经纤维存在弱阳性产物分布。图像分析结果表明,神经元中GnRH受体的相对表达量与其他非神经结构相比差异性显著（P〈0．05）。结果表明,山羊结状神经节中的GnRH受体主要存在于神经元中,山羊结状神经节中的内脏感觉传入神经元对GnRH具有反应性,提示GnRH结状神经节内脏感觉传入神经元的GnRH受体可能作为GnRH对内脏器官的内分泌调节和自主神经对内脏器官的神经调节这2种途径协调作用的节点。     

催产素受体在雌性山羊颈胸神经节的分布

为探讨催产素(OT)是否可以影响雌性山羊颈胸神经节的活动.采用免疫组化SP法观察催产素受体在雌性山 羊颈胸神经节的分布特点.催产素受体在颈胸神经节分布广泛,节内的神经细胞、卫星细胞和过路纤维均有OTR 免疫阳性产物分布;OTR主要在神经细胞中表达,相对表达量与其他非神经结构相比差异性显著(P＜0.05);在神经 细胞中,OTR免疫阳性产物主要存在于胞膜、胞质、核仁,核膜不着色.雌性山羊颈胸神经节神经元对OT具有反应 性,提示OT可能通过影响颈胸神经节神经元的活动,从而经其发出的交感节后神经这一途径调节其所支配的靶器 官如心血管、汗腺、呼吸等的生理活动.     

Immunohistochemical localization of alpha 2-beta 1-glycoprotein in horses

Alpha 2-beta 1-glycoprotein may be found free in horse serum or complexed with alpha-1-proteinase inhibitor to form pre-alpha 2-elastase inhibitor. There has been little information published concerning alpha 2-beta 1-glycoprotein and its possible tissue sources in horses. A peroxidase-antiperoxidase technique was used to identify alpha 2-beta 1-glycoprotein in buffy coat and bone marrow neutrophils of healthy horses. Macrophages and neutrophils in bronchoalveolar lavage samples from clinically normal horses and from horses with chronic pulmonary disease also were positive for alpha 2-beta 1-glycoprotein. Alpha 2-beta 1-glycoprotein was identified in some instances in normal equine hepatocytes of formalin-fixed liver sections. In formalin-fixed lung sections from horses with chronic, small-airway disease and chronic bronchointerstitial pneumonia, alpha 2-beta 1-glycoprotein was observed in some airway secretions and in macrophages.     

Immunohistochemical localization of CB1 receptor in canine salivary glands

CB1 is a member of the G-protein-linked receptor superfamily that is present in the central nervous system as well as in certain peripheral neuronal and non-neuronal tissues. Recently, the presence of CB1 was found in the ductal system of the major salivary glands of laboratory animals, but no data are available for domestic mammals. Thus, in the present study, we examined the presence and distribution of CB1 in the major salivary glands of dogs using immunohistochemical techniques. CB1 was found in the parotid and mandibular glands of adult dogs; positive immunoreaction was localized to the cells of the striated ducts, with a peculiar localization on or near the apical membrane. This particular localization may be explained by the characteristics of this receptor as membrane-associated. The acinar structures were completely negative for CB1. We conclude that CB1 is involved in the control of dog salivary secretion via endogenous substances, likely endocannabinoids. The localization of CB1 highlights that endocannabinoids promote qualitative and/or quantitative changes of the primary saliva in the ductal system.     

青年期广西本地水牛下丘脑催产素的免疫组化定位

为研究青年期广西本地水牛催产素能神经元的表达特点,试验采用免疫组织化学技术中的PicTureTM二步法对广西本地水牛催产素神经元的定位及分布进行了研究.结果表明:下丘脑分泌催产素的神经元主要分布在室旁核、视上核和弓状核,在视前内侧核、视交叉上核、下丘脑前核、下丘脑后核、背内侧核、腹内侧核和乳头体核等核团也有一定数量的阳性神经元;阳性神经纤维仅见于室旁核、弓状核、乳头体核,在正中隆起第三脑室室管膜附近有较多数量的阳性神经纤维.观察结果提示:下丘脑催产素经神经垂体、第三脑室、血管3条途径释放;不同种属动物下丘脑催产素存在差异.     

Immunohistochemical localization of inhibin subunits in the testis of the bull

The differential localization of the inhibin beta subunits betaA and betaB in the testis of adult bull was studied using specific monoclonal and polyclonal primary antibodies. Inhibin betaA- and betaB-subunits were localized only in the Sertoli cells. The inhibin betaA-subunit was observed in the cytoplasm while the betaB-subunit was localized in the nucleus. No specific findings depending on spermatogenic stages were observed among the seminiferous tubules. Moreover, the inhibin alpha-subunit was not detected in the testis of the bulls. In addition, no inhibin subunits were detected in the Leydig cells and spermatogenic cells. These findings indicate the presence of betaA- and betaB-subunits in the bull, which may suggest a possibility that activin is produced and/or stored in the Sertoli cells and regulates spermatogenesis in an autocrine/paracrine manner. Moreover, the inhibin betaB-subunit may be produced in the nucleus but the functional meaning of this is not yet clear.     

Cell-specific localization of the cholecystokininA receptor in the porcine pancreas

Cholecystokinin (CCK) produced in the mucosa of the upper small intestine exerts several biological functions. Its secretion in physiological amounts is modulated by the interaction of extracellular regulators and by binding to intracellular receptors of the target cells. The relative affinity of CCK to its receptor has been characterized in various biological and pharmacological studies and it is now well established that CCK has a higher affinity to the CCKA than to the CCKB receptor. Furthermore CCK influences the secretion of pancreatic enzymes in several species but very little is known about the relationship between CCK and the islet hormone-producing cells in the pig pancreas. The localization of this receptor at the cellular level showed conflicting results in animal studies and has not been described in pigs. The aim of the present study was to characterize the precise cellular location of the CCKA receptor in the porcine pancreas. Polyclonal antiserum was raised against the N-terminal epitope of the CCKA receptor molecule and used for localization studies. Using immunohistochemistry on methanol/acetic acid-fixed, paraffin-embedded pancreas, the CCKA receptor could successfully be localized in islet cells. Parallel staining of serial sections with antibodies directed against insulin and glucagon revealed colocalization with glucagon in alpha cells. No immunoreaction was found in the exocrine pancreas. Our results support the concept that in the porcine species the stimulation of the exocrine pancreas is mediated by the CCKB rather than the CCKA receptor, as it is known for the rat species.     

Immunohistochemical localization of S-100 protein in the pig pituitary gland

Immunohistochemical localization of S-100 protein was studied in anterior, intermediate and posterior lobe of the pig pituitary gland. Two immunopositive cells for S-100 protein were identified: the folliculo-stellate cells (FSc) in the glandular lobes and the pituicytes in the neural lobe. In the anterior lobe, immunoreactive folliculo-stellate cells were scattered among secretory cells. In the area where the secretory cells form strands and follicle-like groups the positive cells were concentrated in groups. In the intermediate lobe, S-100 protein-positive cells were located sparsely among secretory cells and next to secretory follicles and the pituitary cleft. These FSc were more voluminous and displayed fewer cytoplasmic processes. In the neurohypophysis, positive reaction for S-100 protein was seen in the pituicytes. These cells were distributed singly or concentrated in groups. The distribution, and morphologic characteristics of the FSc in the glandular lobes and the pituicytes in the neural lobe in the pig indicate different origin of both S-100 protein-positive cells.     

Immunohistochemical localization of endometrial oestrogen and progesterone receptors in the cow

The presence and distribution of oestrogen and progesterone receptors have been investigated by means of immunohistochemical procedures in the uterus of two groups of cows: the first group underwent superovulatory hormonal treatment while the second group was used as a control group. After the immunohistochemical study no differences regarding the presence and distribution of hormone receptors seemed to be apparent between the two groups, so it was concluded that the topographical distribution and staining intensity of hormone receptors seem to be unaffected by hormonal superovulatory treatment.     

Immunohistochemical localization of chromogranin A in normal tissues from laboratory animals

To analyze the distribution of Chromogranin A in endocrine cells of various species of laboratory animals (dog, gerbil, guinea pig, hamster, monkey, mouse, and fetal, neonatal, and adult rats), normal tissues were stained immunohistochemically with polyclonal anti-bovine Chromogranin A antiserum (SP-1). Selected tissues (pituitary, adrenal, thyroid, parathyroid, pancreas, brain, peripheral nerve, stomach, small and large intestine, bone marrow, spleen, thymus, lymph node, and liver) from these species and from the rabbit were stained with two monoclonal anti-human Chromogranin A antibodies (LK2H10 and PHE5) to compare the immunoreactivities of the monoclonal antibodies and polyclonal antiserum. Staining with the polyclonal antiserum (SP-1) resulted in a broader spectrum of immunoreactivity but had more nonspecific background staining than either monoclonal antibody. Immunoreactivity and staining intensity with SP-1 varied between species, but most endocrine tissues (pituitary cells in the anterior and intermediate lobes, thyroid "C" cells, adrenal medulla, parathyroid, pancreatic islets, and enterochromaffin cells) from most species stained positively. In some species, pancreatic alpha cells stained more intensely, and two populations of adrenal medullary cells with different staining intensities were observed. Sciatic nerve (axonal area) was immunoreactive with monoclonal antibodies and/or the polyclonal antiserum in several species. The spectrum of immunoreactive tissues from fetal and neonatal rats increased with age. There was good cross-reactivity between species with SP-1, but not with either LK2H10 or PHE5. These results indicate that many endocrine cells with secretory granules in laboratory animals express Chromogranin A and that a polyclonal antiserum, such as SP-1, is more sensitive in detecting this protein in various species than monoclonal antibodies such as LK2H10 or PHE5.