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.     

Immunohistochemical localization of insulin-like growth factor-I receptor (IGF-IR) in the developing and mature rat testes

It has been suggested that insulin-like growth factor-I (IGF-I) plays an important role in the regulation of spermatogenesis in the testes. Its signal is mediated predominantly by the IGF-I receptor (IGF-IR). Signalling through IGF-IR has been shown to have a potent survival function. IGF-IR, a transmembrane tyrosine kinase, is widely expressed across many cell types. In this study, we demonstrated the distribution of IGF-IR in testes of differently aged rats. Anti-IGF-IR is a rabbit polyclonal antibody raised against a peptide mapping at the carboxy terminus of the IGF-IR of human origin. Testicular specimens were fixed in Bouin's solution and embedded in paraffin. The paraffin-embedded sections were processed for standard immunohistochemistry by the labelled streptavidin-biotin technique. At postnatal day 19, IGF-IR immunoreactivity was seen moderately in spermatogonia, and slightly both in leptotene and zygotene primary spermatocytes. At postnatal day 35, immunoreactivity was seen slightly both in the pachytene primary spermatocytes and Leydig cells. Although there was intense immunoreactivity in the Leydig cells and in the elongated spermatids on days 50 and 70, the intensity of reaction was decreased in the elongated spermatids in the 10th month. Our results suggest that IGF-IR may play significant roles in testicular function and germ cell development.     

Immunohistochemical identification and fiber type specific localization of protein kinase C isoforms in equine skeletal muscle

OBJECTIVE: To investigate whether protein kinase C (PKC) isoforms are expressed in equine skeletal muscle and determine their distribution in various types of fibers by use of immunofluorescence microscopy. ANIMALS: 5 healthy adult Dutch Warmblood horses. PROCEDURE: In each horse, 2 biopsy specimens were obtained from the vastus lateralis muscle. Cryosections of equine muscle were stained with PKC isoform (alpha, beta1, beta2, delta, epsilon, or zeta)-specific polyclonal antibodies and examined by use of a fluorescence microscope. Homogenized muscle samples were evaluated via western blot analysis. RESULTS: The PKC alpha, beta1, beta2, delta, epsilon, and zeta isoforms were localized within the fibers of equine skeletal muscle. In addition, PKC alpha and beta2 were detected near or in the plasma membrane of muscle cells. For some PKC isoforms, distribution was specific for fiber type. Staining of cell membranes for PKC alpha was observed predominantly in fibers that reacted positively with myosin heavy chain (MHC)-IIa; PKC delta and epsilon staining were more pronounced in MHC-I-positive fibers. In contrast, MHC-I negative fibers contained more PKC zeta than MHC-I-positive fibers. Distribution of PKC beta1 was equal among the different fiber types. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that PKC isoforms are expressed in equine skeletal muscle in a fiber type-specific manner. Therefore, the involvement of PKC isoforms in signal transduction in equine skeletal muscle might be dependent on fiber type.     

Breeding versus survival: proximate causes of abrupt population decline under environmental change in a desert rodent,the midday gerbil (Meriones meridianus)

Studying abrupt ecological shifts under gradual environmental change caused, in particular, by human activity is important for understanding the fundamental aspects and underlying mechanisms of ecological resilience. One of the rare well‐documented examples of an abrupt ecological shift is the delayed step transition of the population of a desert rodent, the midday gerbil (Meriones meridianus), from high‐abundance (1994–2002) to low‐abundance (2003–2017) regimes. This was in response to landscape transformation from desert to steppe caused by the drastic reduction of livestock in the rangelands of southern Russia after the collapse of the USSR in the early 1990s. In this study, we tested whether demographic parameters were correlated with the observed abrupt downward population shift. We found that reproductive activity (the percentage of breeding females, the number of litters, fecundity and the number of young recruited per female) showed no trend over time and did not differ between periods of high and low abundance. In contrast, the adult sex ratio (SR = males: females) decreased significantly with time and was as much as twice more female‐biased for the low‐abundance population regime. However, SR was not related to any reproductive parameter, including the percentage of breeding females. We conclude that proximate reasons for an abrupt population decline in M. meridianus are not associated with the changes in breeding patterns or mate limitation caused by the Allee effect but relate to the increased mortality as a result of the desert landscape being fragmented by steppezation. The mortality is expected to be higher for males as the mobile and dispersing sex.     

Immunohistochemical localization of oestrogen receptor alpha in the various cell categories of chick embryo ovary

The immunohistochemical (IHC) localization of oestrogen receptor alpha (ERα) was studied in the developing left ovary of 14.5-day-old chick embryos. The study was focused in particular on distinguishing in cortex and medulla the different cell categories that proved positive to the reaction, in order to gain further understanding of gonadal cell interactions during ovarian development. Immunostained cells were observed in both the cortex and medulla, but the reactivity for ERα was discontinuous, probably due to variable cell requirements. In the cortex, positivity was observed in cells of the ovarian surface epithelium, in germ cells and in prefollicular cells. In the medulla, positivity was found in the following cell categories: interstitial cells, poorly differentiated somatic cord cells, including those delimiting lacunae, germ cells and their accompanying cells of epithelial origin. Furthermore, the IHC results showed that the intracellular localization of the antigen was cytoplasmic, nuclear, or both. The significance of ERα presence and intracellular localization was discussed in relation and as supplementary to previous research by various Authors. In particular, as regards the unusual cytoplasmic immunoreactivity, a gradual shift of ERα localization from cytoplasmic to nuclear during the embryonic period is suggested.     

Immunohistochemical identification and localization of orexin A and orexin type 2 receptor in the horse gastrointestinal tract

The aim of the present study was to investigate the presence and the distribution of cells containing orexin A and orexin type 2 receptor in the horse stomach and gut, by means of immunohistochemical techniques.Orexin A was identified in the stomach fundic and pyloric regions and in the duodenum. In the same stomach regions, a large subset of orexin A-positive cells also showed orexin type 2 receptor-like immunoreactivity. Moreover, in the duodenum, many of them, seemed to store serotonin.Characteristically, enteric neurons or ganglia also displayed orexin A and, sometimes, orexin type 2 receptor immunoreaction.Orexin A and orexin type 2 receptor immunoreactivity was also found in the nerve fibers in the enteric submucosal layer.Our results, together with data present in the literature, could contribute to the understanding of complex mechanisms regulating the horse gut functionality that are depending very likely on the consequence of the co-operation of both a central and a peripheral control.     

Comparison of localization of the neurokinin 1 receptor with AMPA-type glutamate receptors in the rat spinal cord

With the cloning of the alpha-amino-3-hydroxy-5-methyl-4-isaxole propionic acid (AMPA)-type receptor subunits, it is now possible to localize these receptor subunits in the spinal cord. Comparison of the neurokinin 1 receptor distribution with that of non-N-methyl-D-aspartate glutamate receptor subunits (GluR1-4), considered to be AMPA-type, was investigated in rat spinal cord by immunocytochemical methods. Different patterns of immunolabelling were observed with the antibodies to the GluR1, GluR2/3 and GluR4 subunits in the lumbar spinal cord. Immunolabelling with antibodies to both GluR1 and GluR2/3 revealed intensive staining in the dorsal horn, while staining for GluR2/3 and GluR4 was dense in the motor neurons of the ventral horn. These results suggest that in the rat spinal cord AMPA-type receptors vary their composition according to the region where they are expressed. Neurokinin-1-receptor-expressing neurons in the dorsal horn do not appear to express the GluR4 subunit, however, whether they express the GluR1, GluR2/3 receptors subunits could not be determined.     

Immunohistochemical localization of beta defensins in the endometrium of rat uterus during the postpartum involution period

β-Defensins are small cationic molecules that have antimicrobial actions against bacteria, fungi and viruses and contribute to mucosal immune responses at epithelial sites. The female reproductive tract is an important site of defensin production. This study was conducted to determine the possible changes in proportions and localization of β-defensin 1-4 in the rat uterus at the 1st, 3th, 5th, 10th and 15th days of postpartum and at the period of diestrus using immunohistochemical techniques. In the present study, it was determined that β-defensin 1-4 were generally found in all structural components of the endometrium (luminal and glandular epithelium, stromal cells and blood vessels) in both the nucleus and the cytoplasm of cells during the involution period and diestrus. Suprisingly, immunoreaction of β-defensin 2 was also observed in the lateral membrane of the luminal and glandular epithelial cells on the 10th day of involution and immunostaining of β-defensin 4 was also localized in the apical membrane of the luminal and glandular epithelial cells. The current study demonstrated β-defensin 1-4 immunoreactivities in the endothelium of blood vessels were stronger throughout the involution period. Although β-defensins 2 and 3 were localized in both the nuclei and the cytoplasm of endothelial cells, β-defensins 1 and 4 were present in only cytoplasm. These results show that the most component of rat endometrium expresses human β-defensin 1-4 in a involution-dependent manner. Therefore it may be asserted that these molecules constitute a organised protection to prevent uterus from probable infections during the involution process.     

Immunohistochemical study of caveolin-1 and -2 in the rat retina

The expression of caveolin-1 and -2 in the retina was examined; Western blot analysis showed that both were present. Immunohistochemistry indicated that caveolin-1 was expressed in the majority of retinal layers, including the ganglion cell layer, inner plexiform layer, outer plexiform layer, and in the vascular endothelial cells of the retina. Caveolin-2 was primarily immunostained in the vessels, but in a few other elements as well. This is the first demonstration of caveolin differential expression in the retina of rats, and suggests that caveolin plays an important role in signal transduction in glial cells and neuronal cells.     

Comparison of localization of the neurokinin 1 receptor and nitric oxide synthase with calbindin D labelling in the rat spinal cord

A comparison of the localization of the neurokinin 1 (NK1) receptor and nitric oxide synthase with calbindin D labelling in the lumbar spinal cord was carried out in the rat using immunocytochemistry. Considerable regional variations were observed. Application of the antibody to calbindin D resulted in dense staining in laminae I and II and light staining in the other laminae. Occasional scattered cells were seen in the deep laminae and in the lamina X, the ventral horn and the lateral spinal nucleus. The results indicate that neurones expressing calbindin D, NK1 receptor and NOS are three separate populations in the dorsal horn of the lumbar spinal cord.     

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 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.     

In vitro glucuronidation of the angiotensin II receptor antagonist telmisartan in the cat: a comparison with other species

Ebner, T., Schänzle, G., Weber, W., Sent, U., Elliott, J. In vitro glucuronidation of the angiotensin II receptor antagonist telmisartan in the cat: a comparison with other species. J. vet. Pharmacol. Therap.  36 , 154–160. Glucuronidation of telmisartan comprises nearly its entire metabolic clearance in several mammalian species including human. However, data were lacking for the cat, a species noted for its inability to glucuronidate some drugs. Therefore, the glucuronidation of telmisartan was investigated using feline liver microsomes and compared to liver microsomes of rats, dogs, and human, intestinal human microsomes and cell lines expressing human UDP‐glucuronosyltransferases (UGT). Incubation of telmisartan with cat liver microsomes readily yielded telmisartan glucuronide, and pooled (N = 3 for each gender) cat liver microsomes even showed the highest glucuronidation rate (cat > dog >> human > rat). Michaelis Menten kinetics were observed with K_m of 7.5 and 10 μm and V_max of 3.9 and 3.3 nmol/min/mg for male and female cats, respectively. Confirming the in vitro data, telmisartan glucuronide was detected as the major circulating metabolite in cat plasma. To elucidate which UGT enzymes are involved, telmisartan was incubated with cell lines expressing human UGTs. The highest glucuronidation activity was observed for UGT1A8, UGT1A7, and UGT1A9. In conclusion, telmisartan was effectively glucuronidated in cats. Defects of the UGT1A6 gene in cats do not affect the glucuronidation of telmisartan as it is not a substrate of human UGT1A6.     

Ghrelin在成年皖西白鹅大、小脑内的免疫组化定位

采用免疫组化技术链酶亲和素-生物素-过氧化物酶复合物(StreptAvidin-Biotin-Complex,SABC)法,对成年皖西白鹅大脑和小脑中ghrelin神经元的定位和分布进行了研究。结果显示,大脑分泌ghrelin的神经元和神经纤维主要分布在大脑皮质内,其中锥体层阳性神经元数量最多,多为锥体细胞;小脑分泌ghrelin的神经元主要分布在小脑皮质内,其中蒲肯野细胞层和分子层免疫阳性神经元数量最多,颗粒层阳性神经元数量较少;而大脑和小脑白质内均未见分布。这表明ghrelin在成年皖西白鹅大脑和小脑皮质中分布广泛,可能以自分泌/旁分泌的形式调节中枢神经系统。     

兔圆小囊及肠道组织中抗菌肽的免疫组织化学和免疫电镜细胞化学定位

采用免疫组织化学和免疫电镜细胞化学技术观察了兔圆小囊及其他肠道组织中产抗菌肽细胞的分布定位。免疫组织化学观察发现,产抗菌肽细胞在兔圆小囊的黏膜上皮、圆顶上皮以及淋巴组织的滤泡生发中心、圆顶区和帽区均有分布;在兔感染多杀性巴氏杆菌后,圆小囊及其他肠道组织中的抗菌肽免疫组织化学阳性细胞数量明显增多,阳性反应增强。免疫电镜细胞化学观察结果显示,除肠道黏膜上皮细胞、异嗜性白细胞、巨噬细胞中存在抗菌肽免疫组织化学阳性反应信号外,在淋巴细胞内也发现阳性反应信号,尤其是在圆小囊淋巴组织中的DNES细胞内发现抗菌肽免疫组织化学阳性反应信号。表明圆小囊的DNES细胞和淋巴细胞均可产生抗菌肽。     

Expression of progesterone receptor membrane component 1, serpine mRNA binding protein 1 and nuclear progesterone receptor isoforms A and B in the bovine myometrium during the estrous cycle and early pregnancy

The aim of this study was to investigate the (1) expression of progesterone membrane component 1 (PGRMC1), serpine mRNA binding protein 1 (SERBP1) and progesterone receptor (PR) mRNA and (2) protein expression levels of PGRMC1, SERBP1 and PR isoforms A and B in the bovine myometrium during the estrous cycle and early pregnancy. Uteri from cows on days 1-5, 6-10, 11-16 and 17-21 of the estrous cycle and weeks 3-5, 6-8 and 9-12 of pregnancy were used (n=5-6 per period). There were no changes (P>0.05) in PGRMC1 mRNA expression during the estrous cycle, while expression of SERBP1 and PR mRNA was the lowest (P<0.05) on days 11-16 relative to other days of the cycle. The highest mRNA expression of PGRMC1, SERBP1 and PR was found during pregnancy. There were no changes (P>0.05) in SERBP1 protein expression in cycling and pregnant cows, while the highest (P<0.05) PGRMC1 protein expression was found during weeks 3-5 of pregnancy. Similar protein expression profiles for PRA and PRB were found, and protein levels were highest on days 1-5 of the estrous cycle. From day 6 of the cycle, PRA and PRB protein expression decreased and were maintained at this lower level during pregnancy. In conclusion, our study assessed mRNA and protein expression levels of PGRMC1, SERBP1 and PR in the bovine myometrium during the estrous cycle and the first trimester of pregnancy. It is possible that progesterone (P4) affects myometrial function in a genomic and nongenomic manner.     

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.     

基于MapGIS平台的阿拉善荒漠区啮齿动物群落专家系统中推理机的研究与实现

本研究在前期研发阿拉善荒漠区啮齿动物群落专家系统的基础上,对开垦、轮牧和过牧3种不同类型生境中啮齿动物优势种子午沙鼠(Meriones meridianus)、三趾跳鼠(Dipus sagitta)和小毛足鼠(Phodopus roborovskii)相对数量与8个不同植物因子动态关系进行定量非线性分析,借助MapGIS平台,利用3种优势啮齿动物种群相对数量与8个不同植物因子组成的数据库系统、3种优势啮齿动物种的图像信息组成的知识库系统、3种不同生境下19种不同生态模型组成的模型库系统,构成啮齿动物群落中种群相对数量与植物因子动态关系的推理机,通过程序设计语言VB(Visual Basic)进行MapGIS二次开发,在专家系统中予以设计并实现.     

基于MapGIS平台的阿拉善荒漠区啮齿动物群落专家系统中推理机的研究与实现

本研究在前期研发阿拉善荒漠区啮齿动物群落专家系统的基础上,对开垦、轮牧和过牧3种不同类型生境中啮齿动物优势种子午沙鼠(Meriones meridianus)、三趾跳鼠(Dipus sagitta)和小毛足鼠(Phodopus roborovskii)相对数量与8个不同植物因子动态关系进行定量非线性分析,借助MapGI...