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Xenopus laevis is highly suitable for studying the mechanisms of olfactory reception for water-soluble odorants and for airborne odorants. However, the functional differences of cells and component protein molecules in the olfactory receptors of Xenopus have remained obscure. In recent studies, the patterns of sugar residues expressed on the cell surface have been utilized to analyze the characteristics of neurons, because the sugar chains in neurons play very important roles in targeting and cell-to-cell communication. In this study, we have determined the distribution of sugar residues and glycoproteins in the olfactory receptor organs of Xenopus using lectins as labeling agents, and characterized the receptors of water-soluble odorants and of airborne odorants. The results of lectin histochemical analysis show distributional differences of GlcNAc, GalNAc and mannose between the middle chamber and the lateral chamber of the main nasal cavity. Furthermore, a 65 kDa glycoprotein containing mannose, GlcNAc and GalNAc was specifically detected in the medial chamber of the main cavity epithelium in receptor organs of airborne odorants by SDS-PAGE and lectin blotting. The characteristics of the epithelia demonstrated in this study should further our understanding of the functional differences between the receptors of water-soluble odorants and of airborne odorants at the molecular level.
Anat Rec 1999 08 01
PMID:Characterization of olfactory receptor organs in Xenopus laevis Daudin. 1040 15

It has been postulated that carbohydrates are involved in a variety of cell-cell interactions including blastocyst implantation. In primates, there are only limited investigations on the ultrastructural localisation of the cyclic changes in uterine epithelial surface carbohydrates. Our aim was to investigate such changes during the pre-ovulatory and pre-implantation stages of the reproductive cycle in the marmoset monkey. After fixation of endometrial tissues, avidin-ferritin lectin cytochemistry was employed for apical surface glycan detection at the ultrastructural level. Five lectins were used including Canavalia ensiformis (Con A), Lotus tetragonolobus (LTA), Glycine max (SBA), Phytolacca americana (PWM) and Triticum vulgaris (WGA). Morphometry was used to quantitate changes in the intensity of lectin staining by determining the total number of ferritin particles per unit length of membrane. Surface and intra-cytoplasmic vesicles, stained by the lectins, were also examined. Quantitative ferritin assessment showed that 1 day before presumed implantation (days 11 to 12 after ovulation in the marmoset monkey) there was a significant increase in Con A, LTA and SBA staining on the apical uterine epithelial plasma membrane compared to the pre-ovulatory phase and earlier stages of pregnancy (days 4-8 after ovulation). A significant increase in PWM was also detected from early pregnancy to pre-implantation stages. All lectins except WGA produced reproducible staining within reproductive cycle groups. The greatest variation and intensity of epithelial surface staining was observed with WGA and the weakest with LTA. The patchy staining with LTA compared with thick coverage by WGA indicated the complexity of the carbohydrate arrangement in the glycocalyx of the uterine surface plasma membrane. Reduction of WGA reactivity after neuraminidase treatment suggested that the lectin binding might be related to the presence of heavily sialylated apical uterine membrane glycoconjugates. This is the first high-resolution study in primates to report quantitative cyclic changes in fucosyl, galactosyl, glucosyl, and mannosyl sugar residues of the apical uterine epithelial glycocalyx. The findings support the concept that uterine epithelial glycocalyx surface carbohydrates play a role in preparing a receptive uterine surface.
Anat Rec 1999 07 01
PMID:Ultrastructural studies of glycan changes in the apical surface of the uterine epithelium during pre-ovulatory and and pre-implantation stages in the marmoset monkey. 1041 92

Recently we observed that in human embryos and fetuses with a variety of malformations, not only malformed tissues, but also several non-malformed tissues displayed alterations in the glycosylation pattern. It was the aim of this work to investigate this more or less inexplicable phenomenon under experimental conditions. To this end, we studied a well known mouse model, the mouse mutant undulated, which has an exactly defined genetic defect (substitution in the pax-1 gene) leading to a localized malformation in the vertebral column. The glycosylation pattern was studied using lectin histochemistry. Distribution of binding sites for the lectins RCA I, Con A, SNA, SBA, PNA, LTA and WGA was studied during the organogenesis stages (i.e., days 11-18). It was striking that in mutants, changes in the glycosylation pattern were found not only in the malformed organ (i.e., vertebral anlage), but also in other embryonic tissues, which showed normal morphology. This suggests that the altered glycosylation seems to be a part of genetically determined phenomena throughout the entire organism. Our results show that a defect in a gene with a very restricted expression can cause universal changes in the glycosylation pattern during development.
Anat Rec 2000 03 01
PMID:Extensive glycosylation changes revealed by lectin histochemistry in morphologically normal prenatal tissues of the mouse mutant undulated (un/un). 1070 44

Changes in the patterns of glycosylation of canine haptoglobin have recently been demonstrated by isoelectric focusing and immunoblotting. Fucosylated fractions of haptoglobin were identified by selective binding of a fucose-specific lectin. In this study, similar changes were found in the serum of 86 of 137 dogs with various inflammatory, autoimmune and neoplastic diseases. Major changes, including fucosylation, were observed in 40 of the dogs and were most frequent in association with autoimmune haemolytic anaemia. All 40 cases had markedly increased concentrations of haptoglobin and decreased concentrations of haemoglobin. Minor changes were found in the other 46 dogs, whereas no changes in glycosylation were detected in the serum of 40 healthy dogs.
Vet Rec 2001 Jan 06
PMID:Abnormal microheterogeneity of haptoglobin in serum from dogs with various diseases. 1120 Mar 99

The present histochemical and cytochemical study using a lectin panel (WGA, GSI-A4, GSI-B4, PSA UEA-I, PNA, LCA, Con-A, DBA, MPA, BPA) has demonstrated that, in Podarcis sicula, the differentiation of small follicle cells into pyriform cells by means of intermediate cells is accompanied by the appearance of glycoproteins bearing alpha-GalNAc terminated O-linked side chains on the cell surface. The distribution of DBA- and MPA-binding sites over the follicular epithelium changed during the different stages of oocyte growth. DBA- and MPA-binding sites first appeared at the beginning of folliculogenesis within the zona pellucida (ZP) and on the surface of small cells, i.e., the stem cells of pyriform cells. Afterward, labeling was evident on the cell surfaces of intermediate cells and, later on, also of pyriform cells. On the other hand, no labeling was detected on the small cells located under the basal lamina, which, reportedly, do not differentiate into pyriform cells (Filosa et al. J. Embryol. Exp. Morphol., 1979; 15:297-316). Once pyriform cells were differentiated, the distribution of DBA- and MPA-binding sites over the follicular epithelium remained unchanged until intermediate and pyriform cells underwent apoptosis (Motta et al. J. Exp. Zool., 1996; 276:233-241) and the follicular epithelium transformed into a monolayer composed of small follicle cells only (Filosa Mon. Zool. Ital., 1973; 7:151-165). During this stage of oocyte growth, DBA and MPA labeling gradually decreased to completely disappear in the follicular epithelium of vitellogenic follicles. It is noteworthy that the observed changes in the distribution of DBA- and MPA-binding sites represent the first evidence recognized by lectins of a gradual modification of surface glycoprotein distribution over the follicular epithelium in the ovarian follicles of nonmammalian vertebrates so far studied. Finally, the zona pellucida (ZP), characterized by the presence of GalNAc, GluNAc, Man, and Gal, was demonstrated to be first synthetized by the oocyte and later on by the follicle cells.
Anat Rec 2001 05 01
PMID:Pyriform cell differentiation in Podarcis sicula is accompanied by the appearance of surface glycoproteins bearing alpha-galNAc terminated chains. 1133 65

C2 is a serum glycoprotein that is essential for activation of the classical and lectin pathways of the complement system. We reported previously that in transiently transfected COS cells, C2 accumulates in the endoplasmic reticulum-Golgi intermediate compartment (ERGIC). Transfection with a cDNA corresponding to a variant C2 mRNA in which exon 17 is spliced out, C2Delta(17), resulted in retention of the mutant polypeptide in the ER. We now show that calnexin, a lectin-like chaperone, colocalizes with wild-type (wt) C2 and C2Delta(17). Biosynthetic labeling and sequential immunoprecipitation experiments indicated that colocalization is due to a physical association between calnexin and C2. Immunofluorescence analysis indicated that calnexin was upregulated in cells transfected with either C2 species. Upregulation of calnexin was not affected by castanospermine, which inhibits glucosidases I and II. However, castanospermine inhibited translocation of calnexin to the ERGIC in wt C2 transfected cells. Upregulation of calnexin was also observed in cells transfected with the complement protein factor B, a glycoprotein with extensive structural and functional similarities to C2, but not in cells transfected with complement proteins C3 or factor D, which have no structural similarity to C2, and low or no glycan content, respectively. Calnexin upregulation by transfection with C2 or factor B, but not factor D, was also demonstrated by quantitative analysis of calnexin immunoprecipitates from biosynthetically labeled cells. Increased calnexin expression by overexpressed C2 and factor B appears to be triggered either by the high glycan content of these proteins or, since it also occurs in the presence of castanospermine, by shared features of the structure of these two proteins.
Anat Rec 2002 May 01
PMID:Calnexin is associated with and induced by overexpressed human complement protein C2. 1198 87

The purpose of the present study was to investigate the binding patterns of the plant lectin Bandeiraea simplicifolia I-isolectin B(4) (BSI-B(4)) to sensory neurones in seven mammalian species. The dorsal root ganglia and spinal cords of three rats, mice, guinea pigs, rabbits, flying foxes, cats, and marmoset monkeys were screened for BSI-B(4) using lectin histochemistry. BSI-B(4) binding was associated with the soma of predominantly small-diameter primary sensory neurones in the dorsal root ganglia and their axon terminals within laminae I and II of the superficial dorsal horn in all seven species. The similarities of lectin binding patterns in each of these species suggest that the glycoconjugate to which BSI-B(4) binds has a ubiquitous distribution in mammals, and supports the proposal that this lectin may preferentially bind to a subpopulation of sensory neurones with a similar functional role in each of these species.
Anat Rec 2002 Oct 01
PMID:Analysis of the distribution of binding sites for the plant lectin Bandeiraea simplicifolia I-isolectin B4 on primary sensory neurones in seven mammalian species. 1222 16

The sturgeon is an ancient species of fish that thrives in a wide range of ecological environments, from freshwater to seawater. Basic in this process of adaptation is the ability of the kidney to control fluid filtration and urine formation. However, the morphological basis of this process is mostly unknown. The aim of the present study was to use microdissection techniques (scanning electron microscopy (SEM), transmission electron microscopy (TEM), and lectin-binding histochemistry) to examine the structure of the renal corpuscle of the sturgeon Acipenser nacarii in order to reveal morphologic features that could be related to function, phylogeny, and habitat. The renal corpuscles are aligned along the intrarrenal arteries. The urinary pole shows a siphon-like neck segment (NS) in 92% of the nephrons, whose structural characteristics are different from those of other fish. The podocytes have cuboidal cellular bodies, intercellular contacts, and poorly developed cell processes. The podocyte glycocalyx contains N-acetylglucosamine and lacks sialic acid. The structural and lectin-binding patterns are similar to those found in the immature mammalian kidney. The glomerular basement membrane (GBM) is very thick and consists of three layers: a lamina rara externa, a lamina densa, and a thick subendothelial lamina. The latter contains tubular microfibrils, collagen fibers, and long mesangial cell processes. Frequently, the podocyte bodies attach directly to the GBM, and the area occupied by the filtration slits is very small. Furthermore, the GBM shows a glycosylation pattern different from that observed in most vertebrates. Contrary to what would be expected in sturgeons living in freshwater, the A. nacarii renal corpuscle morphology suggests a low glomerular filtration rate.
Anat Rec A Discov Mol Cell Evol Biol 2003 Jun
PMID:Renal corpuscle of the sturgeon kidney: an ultrastructural, chemical dissection, and lectin-binding study. 1274 Sep 51

The heart extracellular matrix protein hLAMP-1 (lectin-associated matrix protein in the heart) is a component of the particulate matrix that activates the AV endothelium prior to its transformation into mesenchyme within the atrioventricular canal and proximal outflow tract of the heart. The role of hLAMP-1 in this process has yet to be determined, in part because of the limited amount of material available for analysis. To overcome this liability, a monoclonal antibody to hLAMP-1 has been used to recognize proteins expressed by cDNA clones. The isolated cDNAs encode an mRNA consistent with previously published immunohistochemical results. Expression profiles of these clones by in situ hybridization revealed staining in areas of the heart that expressed hLAMP-1 by immunocytochemistry. Taken together, these results suggest that these clones, which represent an expressed sequence tag for hLAMP-1, should provide the basis for isolating a full-length cDNA of hLAMP-1, a prerequisite for determining the functional role of this protein in heart development.
Anat Rec A Discov Mol Cell Evol Biol 2004 Apr
PMID:Identification of cDNA clones that encode hLAMP-1, a component of the particulate matrix associated with cardiac mesenchyme formation. 1505 58

Hyperplasia and squamous metaplasia of the prostatic epithelium are conditions induced by oestrogens. Diethylstilbestrol (DES) has been banned from cattle used for beef production because of the health risks. The potential use of molecular markers for the detection of illegal oestrogen administration was evaluated by taking samples of prostatic tissue from control bullocks, bullocks which had been treated with oestrogens, and bullocks sacrificed 21 and 90 days after a single dose of DES. The expression of the glycoconjugates was examined by lectinhistochemistry and the lectin binding pattern was characterised in epithelium and connective tissue. In the animals sacrificed after 21 days there was an increase in the binding of one lectin (JAC) and there was an increase in the binding of one of the other lectins (DBA) in the animals sacrificed after 90 days. An increase in SWGA lectin staining was observed in the bullocks that had probably been treated with oestrogen and in the animals sacrificed 90 days after the inoculation with DES. There were also differences between the binding of SWGA in the control bullocks and the other groups.
Vet Rec 2004 Mar 06
PMID:Lectin binding patterns in hyperplastic and metaplastic bullock prostate tissues after diethylstilbestrol administration. 1505 37


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