Showing papers on "Apical cytoplasm published in 2014"

Mutations in CCNO result in congenital mucociliary clearance disorder with reduced generation of multiple motile cilia

Abstract: Using a whole-exome sequencing strategy, we identified recessive CCNO (encoding cyclin O) mutations in 16 individuals suffering from chronic destructive lung disease due to insufficient airway clearance. Respiratory epithelial cells showed a marked reduction in the number of multiple motile cilia (MMC) covering the cell surface. The few residual cilia that correctly expressed axonemal motor proteins were motile and did not exhibit obvious beating defects. Careful subcellular analyses as well as in vitro ciliogenesis experiments in CCNO-mutant cells showed defective mother centriole generation and placement. Morpholino-based knockdown of the Xenopus ortholog of CCNO also resulted in reduced MMC and centriole numbers in embryonic epidermal cells. CCNO is expressed in the apical cytoplasm of multiciliated cells and acts downstream of multicilin, which governs the generation of multiciliated cells. To our knowledge, CCNO is the first reported gene linking an inherited human disease to reduced MMC generation due to a defect in centriole amplification and migration.

RAB26 coordinates lysosome traffic and mitochondrial localization

Abstract: As they mature, professional secretory cells like pancreatic acinar and gastric chief cells induce the transcription factor MIST1 (also known as BHLHA15) to substantially scale up production of large secretory granules in a process that involves expansion of apical cytoplasm and redistribution of lysosomes and mitochondria. How a scaling factor like MIST1 rearranges cellular architecture simply by regulating expression levels of its transcriptional targets is unknown. RAB26 is a MIST1 target whose role in MIST1-mediated secretory cell maturation is also unknown. Here, we confirm that RAB26 expression, unlike most Rabs which are ubiquitously expressed, is tissue specific and largely confined to MIST1-expressing secretory tissues. Surprisingly, functional studies showed that RAB26 predominantly associated with LAMP1/cathepsin D lysosomes and not directly with secretory granules. Moreover, increasing RAB26 expression – by inducing differentiation of zymogen-secreting cells or by direct transfection – caused lysosomes to coalesce in a central, perinuclear region. Lysosome clustering in turn caused redistribution of mitochondria into distinct subcellular neighborhoods. The data elucidate a novel function for RAB26 and suggest a mechanism for how cells could increase transcription of key effectors to reorganize subcellular compartments during differentiation.

Hepatoma-derived growth factor: from the bovine uterus to the in vitro embryo culture

Abstract: Early in cow embryo development, hepatoma-derived growth factor (HDGF) is detectable in uterine fluid. The origin of HDGF in maternal tissues is unknown, as is the effect of the induction on developing embryos. Herein, we analyze HDGF expression in day 8 endometrium exposed to embryos, as well as the effects of recombinant HDGF (rHDGF) on embryo growth. Exposure to embryos did not alter endometrial levels of HDGF mRNA or protein. HDGF protein localized to cell nuclei in the luminal epithelium and superficial glands and to the apical cytoplasm in deep glands. After uterine passage, levels of embryonic HDGF mRNA decreased and HDGF protein was detected only in the trophectoderm. In fetal fibroblast cultures, addition of rHDGF promoted cell proliferation. In experiments with group cultures of morulae in protein-free medium containing polyvinyl alcohol, adding rHDGF inhibited blastocyst development and did not affect cell counts when the morulae were early (day 5), whereas it enhanced blastocyst development and increased cell counts when the morulae were compact (day 6). In cultures of individual day 6 morulae, adding rHDGF promoted blastocyst development and increased cell counts. Our experiments with rHDGF indicate that the growth factor stimulates embryonic development and cell proliferation. HDGF is synthesized similarly by the endometrium and embryo, and it may exert embryotropic effects by autocrine and/or paracrine mechanisms.

Increased Toll-like receptor 5 expression indicates esophageal columnar dysplasia

Abstract: Toll-like receptor 5 (TLR5) is an immune receptor, which recognizes bacterial flagellin. Increased expression has been reported in various premalignant and malignant lesions indicating a role in carcinogenesis. We assessed the expression of TLR5 in normal esophageal squamous epithelium, Barrett’s esophagus with and without dysplasia, and in esophageal adenocarcinoma. Specimens with normal esophagus (n = 93), gastric (n = 75) or intestinal metaplasia (n = 53) without dysplasia, and low-grade (n = 56) or high-grade dysplasia (n = 33) and esophageal adenocarcinoma (n = 94) were studied. TLR5 immunohistochemical stainings were analyzed for the proportion of positive cells and the intensity of expression. In normal squamous epithelium, only the basal third showed TLR5 expression. In esophageal gastric or intestinal metaplasia, expression was present in majority of the cells but significantly weaker (p < 0.001) than in dysplastic epithelium. In dysplasia, expression extended to the apical cytoplasm, contrasting basolateral expression in non-dysplastic columnar epithelium. Receiver operating characteristic curve analysis showed that moderate to high expression intensity of TLR5 indicates low-grade dysplasia with 86 % sensitivity and 83 % specificity. Carcinomas showed increased expression in comparison with non-dysplastic columnar epithelium, but there was no association with prognosis. Our results indicate that the esophageal columnar dysplasia is associated with clear increase of TLR5 expression and dissolution of regular polarized expression. TLR5 staining provides a possible biomarker for the recognition of low-grade dysplasia. In addition, the findings suggest a role for abnormal expression of TLR5 in the pathogenesis of esophageal adenocarcinoma and suggest importance of altered microbiome in the pathogenesis of complications of Barrett’s esophagus.

Ductuli efferentes of the male Golden Syrian hamster reproductive tract

Abstract: Efferent ductules are responsible for the transportation of spermatozoa from the testis to the epididymis and their epithelium is responsible for the reabsorption of over 90% of the luminal fluid. The purpose of this research was to characterize the gross morphology and histology of efferent ductules in the male Golden Syrian hamster. The efferent ductules emerge from rete testis with a unique polarity at the apex or cephalic pole of the testis. The number of efferent ductules varied from 3 to 10 with an average of 6.0 and blind ending ducts were observed in approximately 56% of the males. The ductules merged into a single common duct prior to entering the caput epididymidis. The proximal efferent ductule lumen was wider than the distal (conus and common ducts), consistent with reabsorption of most of the luminal fluid, as was morphology of the ductal epithelium. Non-ciliated cells in the proximal region had prominent endocytic apparatuses, showing both coated pits and apical tubules in the apical cytoplasm. Large basolateral, intercellular spaces were also present in the epithelium of the proximal region. Distal non-ciliated cells had an abundance of large endosomes and lysosomal granules. Localisation of sodium/hydrogen exchanger-3 (NHE3; SLC9A3) and aquaporins 1 and 9 (AQP1, AQP9) along the microvillus border was also consistent with ion transport and fluid reabsorption by this epithelium. In comparison, the caput epididymidis epithelium expressed only AQP9 immunostaining. Another unusual feature of the hamster efferent ductules was the presence of glycogen aggregates in the basal cytoplasm of small groups of epithelial cells, but only in the proximal ducts near the rete testis. Androgen (AR), estrogen (ESR1 and ESR2) and vitamin D receptors (VDR) were also abundant in epithelial nuclei of proximal and distal efferent ductules. In comparison, caput epididymidis showed very little immunostaining for ESR1.

Three dimensional cultures: a tool to study normal acinar architecture vs. malignant transformation of breast cells.

Abstract: Invasive breast carcinomas are a group of malignant epithelial tumors characterized by the invasion of adjacent tissues and propensity to metastasize. The interplay of signals between cancer cells and their microenvironment exerts a powerful influence on breast cancer growth and biological behavior1. However, most of these signals from the extracellular matrix are lost or their relevance is understudied when cells are grown in two dimensional culture (2D) as a monolayer. In recent years, three dimensional (3D) culture on a reconstituted basement membrane has emerged as a method of choice to recapitulate the tissue architecture of benign and malignant breast cells. Cells grown in 3D retain the important cues from the extracellular matrix and provide a physiologically relevant ex vivo system2,3. Of note, there is growing evidence suggesting that cells behave differently when grown in 3D as compared to 2D4. 3D culture can be effectively used as a means to differentiate the malignant phenotype from the benign breast phenotype and for underpinning the cellular and molecular signaling involved3. One of the distinguishing characteristics of benign epithelial cells is that they are polarized so that the apical cytoplasm is towards the lumen and the basal cytoplasm rests on the basement membrane. This apico-basal polarity is lost in invasive breast carcinomas, which are characterized by cellular disorganization and formation of anastomosing and branching tubules that haphazardly infiltrates the surrounding stroma. These histopathological differences between benign gland and invasive carcinoma can be reproduced in 3D6,7. Using the appropriate read-outs like the quantitation of single round acinar structures, or differential expression of validated molecular markers for cell proliferation, polarity and apoptosis in combination with other molecular and cell biology techniques, 3D culture can provide an important tool to better understand the cellular changes during malignant transformation and for delineating the responsible signaling.

Histochemical and Immunohistochemical Analysis of the Stomach of Rhinella icterica (Anura, Bufonidae)

Abstract: The stomach of Rhinella icterica was analyzed at light microscopy, employing histochemical techniques, lectin histochemistry, and immunohistochemistry for identifying enteroendocrine cells (EC). Although the stomach was composed of fundic and pyloric regions, its wall is formed by mucosa, submucosa, muscularis, and serosa. The mucosa was lined by a simple columnar mucous epithelium, supported by loose connective tissue. Several tubular, simple glands were composed of mucous neck cells, containing oxynticopeptic cells and EC cells. The mucous neck cells were rich in neutral glycoconjugates. The oxynticopeptic cells were predominant in fundic glands, exhibiting weaker alcianophilic reaction at their apical cytoplasm. Serotonin (5-HT) immunoreactive (IR) cells occurred throughout the entire stomach, preferentially located among mucous cells at upper part of the fundic glands. The muscularis mucosae, formed of smooth muscle, separated the mucosal layer from the submucosa, both of which were constituted by loose connective tissue, but without glands. Lymphoid modules occurred in the mucosa at the boundary at the stomach and the gut. In addition, the muscularis was constituted by two sublayers, the circular internal and the longitudinal external, being recovered by the connective tissue of the serosa.

The Stomach of the Adult African Catfish (Clarias gariepinus, Siluriformes: Clariidae) in Farm Conditions: A Morphological and Mucin Histochemistry Analysis

Abstract: Se efectuo un estudio de tipo histologico e histoquimico con mucina, en el estomago del bagre africano (Clarias gariepinus, Siluriformes: clariidae) cultivado en granja acuicola, para establecer su anatomia basica, debido a la escasa informacion disponible en la literatura La investigacion genero datos para ulteriores investigaciones, que contribuiran con los ictiopatologos y ayudaran a los nutricionistas en el manejo alimentario Despues de su cultivo en una granja acuicola comercial, los peces aparentemente sanos, fueron inmovilizados y sacrificados, usando cloroformo Se tomaron muestras de las regiones cardial, fundica y pilorica, y se procesaron histologicamente En su conjunto, el estomago presentaba una apariencia de “J” El esofago estaba situado cranealmente al estomago, pero no se observo un esfinter que los separara Caudalmente, un esfinter pilorico separaba al estomago del intestino proximal El estomago contenia tres regiones: cardiaca, fundica y pilorica La totalidad de la superficie estomacal se encontraba recubierta por un epitelio cilindrico, que contenia mucina en el citoplasma apical Solamente las regiones cardiaca y fundica tenian glandulas gastricas en la lamina propia, que eran mas desarrolladas en la region fundica La tunica muscular poseia celulas musculares lisas con unaorientacion interna circular y externa longitudinal La histoquimica mediante la mucina revelo la presencia solo de mucina neutra en el estomago Los resultados obtenidos en este estudio sugieren que la ausencia de glandulas gastricas en el piloro, pudiera ser el resultado de una adaptacion de esta especie para reducir la cantidad de acido que penetra en el intestino proximal; por tanto, pudiera ayudar al medio alcalino a maximizar las acciones de las enzimas pancreaticas en el intestino proximal La sola presencia de mucina neutra en el estomago actuaria como amortiguador contra el elevado contenido acido del estomago Las mucinas neutras tambien estan asociadas con el transporte de micromoleculas Abstract The stomach of the farmed African catfish (Clarias gariepinus, Siluriformes: clariidae) was investigated in this study to establish its basic anatomy, as there is scanty information available on the literature It produced to baseline data for further investigative research, help fish clinicians in pathology and aid nutritionists in feed management After cropping from a commercial aquaculture, apparently healthy fish were immobilized and euthanized by using chloroform The samples were dissected and slices of cardiac, fundic and pyloric stomach processed through routine histological procedures Grossly, the stomach was J-shape Cranial to the stomach was the oesophagus but no sphincter was seen separating them Caudally, a pyloric sphincter separated the stomach from the proximal intestine The stomach contained three regions cardiac, fundic and pyloric The entire stomach surface was lined by a simple columnar epithelium containing mucin at the apical cytoplasm Only the cardiac and fundic regions contained gastric glands in the lamina propria, but the glands were more developed in the fundic region The tunica muscularis contained smooth muscle cells in an inner circular and outer longitudinal orientation Mucin histochemistry revealed the presence of only neutral mucin in the stomach The results obtained in this study suggest that the absence of gastric glands in the pylorus may be an adaptation of this species to reduce the quantity of acid entering the proximal intestine; hence it might help the alkaline medium to maximize pancreatic enzyme actions in the proximal intestine The presence of only neutral mucin in the stomach will act as a buffer to neutralize the high acidic stomach content Neutral mucins are also associated with transport of macromolecules

The expression and cellular localization of galectin-1 and galectin-3 in the Fallopian tube are altered in women with tubal ectopic pregnancy

Abstract: Galectin-1 and galectin-3 are abundantly expressed at implantation sites in the uterus, suggesting their involvement in the establishment of pregnancy. In this study, we examined the expression and localization of galectin-1 and galectin-3 in fallopian tubes from nonpregnant women, and in those presenting with tubal ectopic pregnancy. There was no significant difference in the expression of either galectin-1 (LGALS1) or galectin-3 (LGALS3) transcripts in the fallopian tube across the menstrual cycle. Their expressions in the fallopian tube were inversely correlated to each other (r = -0.5134, p < 0.0001) and differentially localized. Galectin-1 protein was abundant in the stroma of nonpregnant fallopian tubes, whereas galectin-3 was mainly localized to the epithelium, notably to the cilia of ciliated cells and the apical cytoplasm of secretory cells. In ectopic pregnancies, LGALS3 expression was significantly reduced (p < 0.0001), but LGALS1 expression did not change when compared to nonpregnant fallopian tubes collected during the mid-secretory phase. The percentage of fallopian tube epithelial cells expressing galectin-3 in cilia tended to be reduced (p = 0.0685), with an accompanying loss of a normal ciliary structure, while nuclear galectin-3 increased (p < 0.05) in ectopic pregnancies. Epithelial immunostaining for galectin-1 tended to be elevated in fallopian tubes from women with ectopic pregnancy. Coculture of human trophoblast origin SW71 cells significantly increased LGALS1 expression in human fallopian tube epithelial OE-E6/E7 cells, suggesting that trophoblast-derived products regulate LGALS1 expression in the oviductal epithelium. These findings imply a differential contribution of galectin-1 and galectin-3 in the homeostasis of human fallopian tubes and in the pathophysiology of ectopic pregnancy.

Biodistribution and Functional Reactivity of Some Immune Cells in Peyer's Patches , Cecal Patches and Spleen of Albino Mice after Treatment with the Nanoparticles

Abstract: Gut-associated lymphoid tissues (GALT) including Peyer's patches (PPs) and cecal patches (CPs) have a vital role in the induction of immune response against pathogens. Spleen is considered as non-GALT; it has a variety of immune cells that support the immune response. In this study, the expression of alpha L-fucose residues and biodistribution of Lectin Ulex Europaeus Agglutinin I (UEA I+) cells, IgA+ cells, 33D1+ dendritic cells (DCs) and CD11b+ macrophage subsets in the GALT (PPs and CPs) and in non-GALT (spleen) were investigated using UEAI lectin histochemistry and immunohistochemistry. The immune response induction of alpha L-fucose residue, UEAI+ cells, IgA+ cells, 33D1+ DCs and CD11+ macrophages subsets also will be investigated after treatment with amino-conjugated fluorescent silica nanoparticles (FSNPs). After treatment with FSNPs an induction of alpha L-fucose residue was observed in the PPs M cells in all cytoplasm, rather than the apical cytoplasm as shown in the control group. Also, the number of UEAI+ cells and IgA+ cells in PPs sub-epithelial dome (SED) increased 4 times and 2 times higher, respectively, as compared with that of the control group. The CD11b+ macrophages, but not 33D1+ DCs subsets located in PPs SED, had the main role to uptake FSNPs. In the case of spleen, the number of 33D1+ DCs and CD11b+ macrophages subsets was 30 times and 25 times higher, respectively, as compared with that of control group. These data indicated that alpha L-fucose residue and IgA+ cells, 33D1+ DCs and CD11b+ macrophages subsets in PPs as GALT and spleen as non-GALT had important role, but in different ratios, in the immune system induction against FSNPs.

El Estómago del Bagre Africano Adulto (Clarias gariepinus, Siluriformes: Clariidae) en Condiciones de Granja: Análisis Morforlógico e Histoquímico por Mucina

Abstract: The stomach of the farmed African catfish (Clarias gariepinus, Siluriformes: clariidae) was investigated in this study to establish its basic anatomy, as there is scanty information available on the literature. It produced to baseline data for further investigative research, help fish clinicians in pathology and aid nutritionists in feed management. After cropping from a commercial aquaculture, apparently healthy fish were immobilized and euthanized by using chloroform. The samples were dissected and slices of cardiac, fundic and pyloric stomach processed through routine histological procedures. Grossly, the stomach was J-shape. Cranial to the stomach was the oesophagus but no sphincter was seen separating them. Caudally, a pyloric sphincter separated the stomach from the proximal intestine. The stomach contained three regions cardiac, fundic and pyloric. The entire stomach surface was lined by a simple columnar epithelium containing mucin at the apical cytoplasm. Only the cardiac and fundic regions contained gastric glands in the lamina propria, but the glands were more developed in the fundic region. The tunica muscularis contained smooth muscle cells in an inner circular and outer longitudinal orientation. Mucin histochemistry revealed the presence of only neutral mucin in the stomach. The results obtained reSumen

Tufting enteropathy with EpCAM mutation: case report Enteropatia com formação de tufos epiteliais e mutação do gene EpCAM: relato de caso

Abstract: Tufting enteropathy (TE), also known as intestinal epithelial dysplasia (IED), is a rare congenital enteropathy related to an early-onset of severe intractable diarrhea due to specific abnormalities of the intestinal epithelium and mutations of the EpCAM gene. TE is characterized by clinical and histological heterogeneity, such as with low or without mononuclear cell infiltration of the lamina propria, and abnormalities of basement membrane. TE can be associated with malformations, other epithelial diseases, or to abnormal enterocytes development and/or differentiation. The authors report a case of a Brazilian child with TE associated with c.556-14A>G mutation in the EpCAM gene (NM_002354.2).Key words: tufting enteropathy; intractable diarrhea; intestinal mucosa; epithelial cell adhesion molecule; EpCAM gene. introDuCtion Tufting enteropathy (TE), also known as intestinal epithelial dysplasia (IED), is a rare disorder that causes chronic watery diarrhea, and is characterized by the presence of focal epithelial “tufts” composed of clusters of closely packed enterocytes with round, teardrop-shaped projections in the apical cytoplasm. TE is present in the first few months of life, and most patients require total parenteral nutrition to acquire adequate caloric and fluid intake for normal growth and development