Expression and localization of aquaporins in rat gastrointestinal tract

1999 ◽  
Vol 276 (3) ◽  
pp. C621-C627 ◽  
Author(s):  
Yu Koyama ◽  
Tadashi Yamamoto ◽  
Tatsuo Tani ◽  
Kouei Nihei ◽  
Daisuke Kondo ◽  
...  
Keyword(s):  
Small Intestine ◽  
In Situ Hybridization ◽  
Gastrointestinal Tract ◽  
Epithelial Cells ◽  
Cellular Localization ◽  
Basolateral Membrane ◽  
Rnase Protection Assay ◽  
Rnase Protection ◽  
Basolateral Membranes

A family of water-selective channels, aquaporins (AQP), has been demonstrated in various organs and tissues. However, the localization and expression of the AQP family members in the gastrointestinal tract have not been entirely elucidated. This study aimed to demonstrate the expression and distribution of several types of the AQP family and to speculate on their role in water transport in the rat gastrointestinal tract. By RNase protection assay, expression of AQP1–5 and AQP8 was examined in various portions through the gastrointestinal tract. AQP1 and AQP3 mRNAs were diffusely expressed from esophagus to colon, and their expression was relatively intense in the small intestine and colon. In contrast, AQP4 mRNA was selectively expressed in the stomach and small intestine and AQP8 mRNA in the jejunum and colon. Immunohistochemistry and in situ hybridization demonstrated cellular localization of these AQP in these portions. AQP1 was localized on endothelial cells of lymphatic vessels in the submucosa and lamina propria throughout the gastrointestinal tract. AQP3 was detected on the circumferential plasma membranes of stratified squamous epithelial cells in the esophagus and basolateral membranes of cardiac gland epithelia in the lower stomach and of surface columnar epithelia in the colon. However, AQP3 was not apparently detected in the small intestine. AQP4 was present on the basolateral membrane of the parietal cells in the lower stomach and selectively in the basolateral membranes of deep intestinal gland cells in the small intestine. AQP8 mRNA expression was demonstrated in the absorptive columnar epithelial cells of the jejunum and colon by in situ hybridization. These findings may indicate that water crosses the epithelial layer through these water channels, suggesting a possible role of the transcellular route for water intake or outlet in the gastrointestinal tract.

Integrin alpha subunit mRNAs are differentially expressed in early Xenopus embryos

Development ◽  
1993 ◽  
Vol 117 (4) ◽  
pp. 1239-1249 ◽  
Author(s):  
C.A. Whittaker ◽  
D.W. DeSimone
Keyword(s):  
In Situ Hybridization ◽  
Rnase Protection Assay ◽  
Early Embryo ◽  
Mrna Levels ◽  
Rnase Protection ◽  
Dorsal Midline ◽  
Transmembrane Receptors ◽  
Early Embryos ◽  
Whole Mount

Adhesion of cells to extracellular matrix proteins is mediated, in large part, by transmembrane receptors of the integrin family. The identification of specific integrins expressed in early embryos is an important first step to understanding the roles of these receptors in developmental processes. We have used polymerase chain reaction methods and degenerate oligodeoxynucleotide primers to identify and clone Xenopus integrin alpha subunits from neurula-stage (stage 17) cDNA. Partial cDNAs encoding integrin subunits alpha 2, alpha 3, alpha 4, alpha 5, alpha 6 and an alpha IIb-related subunit were cloned and used to investigate integrin mRNA expression in early embryos by RNase protection assay and whole-mount in situ hybridization methods. Considerable integrin diversity is apparent early in development with integrins alpha 2, alpha 3, alpha 4, alpha 5 and alpha 6 each expressed by the end of gastrulation. Both alpha 3 and alpha 5 are expressed as maternal mRNAs. Zygotic expression of alpha 2, alpha 3, alpha 4 and alpha 6 transcripts begins during gastrulation. Integrin alpha 5 is expressed at relatively high levels during cleavage, blastula and gastrula stages suggesting that it may represent the major integrin expressed in the early embryo. We demonstrated previously that integrin beta 1 protein synthesis remains constant following induction of stage 8 animal cap cells with activin (Smith, J. C., Symes, K., Hynes, R. O. and DeSimone, D. W. (1990) Development 108, 289–298.). Here we report that integrin alpha 3, alpha 4 and alpha 6 mRNA levels increase following induction with 10 U/ml activin-A whereas alpha 5, beta 1 and beta 3 mRNA levels remain unchanged. Whole-mount in situ hybridization reveals that alpha 3 mRNAs are expressed by cells of the involuting mesoderm in the dorsal lip region of early gastrulae. As gastrulation proceeds, alpha 3 expression is localized to a stripe of presumptive notochordal cells along the dorsal midline. In neurulae, alpha 3 mRNA is highly expressed in the notochord but becomes progressively more restricted to the caudalmost portion of this tissue as development proceeds from tailbud to tadpole stages. In addition, alpha 3 is expressed in the forebrain region of later stage embryos. These data suggest that integrin-mediated adhesion may be involved in the process of mesoderm involution at gastrulation and the organization of tissues during embryogenesis.

scholarly journals Synthesis of transthyretin (pre-albumin) mRNA in choroid plexus epithelial cells, localized by in situ hybridization in rat brain.

1986 ◽  
Vol 34 (7) ◽  
pp. 949-952 ◽  
Author(s):  
A J Stauder ◽  
P W Dickson ◽  
A R Aldred ◽  
G Schreiber ◽  
F A Mendelsohn ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Epithelial Cells ◽  
Rat Brain ◽  
Choroid Plexus ◽  
Cellular Localization ◽  
Lateral Ventricles ◽  
The Brain ◽  
Albumin Mrna ◽  
Choroid Plexus Epithelial

The sites of synthesis of transthyretin in the brain were investigated using in situ hybridization with [35S]-labeled recombinant cDNA probes specific for transthyretin mRNA. Autoradiography of hybridized coronal sections of rat brain revealed specific cellular localization of transthyretin mRNA in choroid plexus epithelial cells of the lateral, third, and fourth ventricles. Transferrin mRNA was also investigated and, in contrast to transthyretin mRNA, was localized mainly in the lateral ventricles. Our results indicate that substantial synthesis of transthyretin and transferrin mRNA may occur in the choroid plexus.

Cellular localization of mRNAs for prostaglandin E receptor subtypes in mouse gastrointestinal tract

1997 ◽  
Vol 272 (3) ◽  
pp. G681-G687 ◽  
Author(s):  
K. Morimoto ◽  
Y. Sugimoto ◽  
M. Katsuyama ◽  
H. Oida ◽  
K. Tsuboi ◽  
...  
Keyword(s):  
Gastrointestinal Tract ◽  
Epithelial Cells ◽  
Cellular Localization ◽  
The Body ◽  
Prostaglandin E ◽  
Receptor Subtypes ◽  
Fundic Gland ◽  
Intestinal Functions ◽  
Myenteric Ganglia

Regional and cellular distribution of mRNAs for prostaglandin E (PGE) receptor subtypes was investigated in the mouse gastrointestinal tract by in situ hybridization. Strong signals for EP1 transcripts were detected in cells of the muscularis mucosae layer, especially in the body of the stomach. Intense signals for EP3 transcripts were detected in neurons of the myenteric ganglia throughout the tract. Moderate EP3 mRNA expression was also observed in fundic gland epithelial cells, except for surface mucous cells in the stomach. Expression of EP4 mRNA was moderate in surface epithelial cells of the corpus and in glands from the surface to the base of the antrum. Strong EP4 signals were observed in the epithelium in the duodenum, jejunum, and ileum. In the ileum, signals were only observed in the upper part of the villi. However, no or weak signals for EP2 transcripts were detected. These findings suggest that PGE2 modulates various gastric or intestinal functions via at least three different PGE receptors.

scholarly journals Regional and maturation-associated expression of endothelin 2 in rat gastrointestinal tract.

1995 ◽  
Vol 43 (2) ◽  
pp. 203-209 ◽  
Author(s):  
S M de la Monte ◽  
T Quertermous ◽  
C C Hong ◽  
K D Bloch
Keyword(s):  
Gene Expression ◽  
Gastrointestinal Tract ◽  
Stromal Cells ◽  
Rnase Protection Assay ◽  
Secretory Activity ◽  
Intestinal Epithelial ◽  
Rnase Protection ◽  
Adult Rat ◽  
Vasoactive Peptides

Endothelin 2 (ET2), also referred to as vasoactive intestinal contractor peptide, is a member of a family of vasoactive peptides. ET2 is a potent constrictor of intestinal smooth muscle, and the mRNA that encodes it has been detected in murine intestinal extracts. To further investigate the potential physiological roles of ET2, we characterized the cellular distribution of ET2 gene expression in adult rat gastrointestinal tract. Using an RNAse protection assay, an overall proximal to distal gradient of increasing ET2 gene expression was observed from stomach to colon. In situ hybridization studies confirmed this finding and demonstrated ET2 mRNA localized in lamina propria stromal cells. Moreover, ET2 gene expression in stromal cells increased from crypt to villous tip. The results demonstrate that ET2 is produced by stromal cells in villi throughout the intestine. Increased ET2 gene expression at the villous tip is associated with more mature overlying epithelial cells, suggesting a possible role for this vasoactive peptide in intestinal epithelial differentiation or secretory activity.

scholarly journals Localization of amidating enzymes (PAM) in rat gastrointestinal tract.

1993 ◽  
Vol 41 (11) ◽  
pp. 1617-1622 ◽  
Author(s):  
A Martínez ◽  
M A Burrell ◽  
M Kuijk ◽  
L M Montuenga ◽  
A Treston ◽  
...  
Keyword(s):  
Small Intestine ◽  
In Situ Hybridization ◽  
Gastrointestinal Tract ◽  
Digestive Tract ◽  
Endocrine Cells ◽  
Regulatory Peptides ◽  
Serial Sections ◽  
Gastric Glands ◽  
Amidating Enzymes

We studied the distribution of the two enzymes involved in post-translational C-terminal alpha-amidation of regulatory peptides in rat digestive tract, using immunocytochemical methods and in situ hybridization techniques. The enzymes were located in most of the fibers and neurons of the myenteric and submucous plexus throughout the entire digestive tract and in endocrine cells of the stomach and colon. Staining of reverse-face serial sections demonstrated that the enzymes in endocrine cells of the stomach co-localized with gastrin in the bottom of the gastric glands. Some gastrin-immunoreactive cells near the neck of the gland were negative for PAM, suggesting that amidation takes place only in the more mature cells. In the colon all cells immunoreactive for glucagon and GLP1 were also positive for peptidylglycine alpha-hydroxylating monooxygenase (PHM) but not for peptidyl-alpha-hydroxyglycine alpha-amidating lyase (PAL). The absence of immunoreactivity for the amidating enzymes in endocrine cells of the small intestine, known to produce C-terminally amidated peptides, suggests the existence of other amidating enzymes.

scholarly journals Localization of Cytochrome P450 CYP2S1 Expression in Human Tissues by In Situ Hybridization and Immunohistochemistry

2005 ◽  
Vol 53 (5) ◽  
pp. 549-556 ◽  
Author(s):  
Sirkku T. Saarikoski ◽  
Harriet A.-L. Wikman ◽  
Gillian Smith ◽  
C. Henrik J. Wolff ◽  
Kirsti Husgafvel-Pursiainen
Keyword(s):  
Cytochrome P450 ◽  
In Situ Hybridization ◽  
Epithelial Cells ◽  
Cellular Localization ◽  
Tumor Development ◽  
Tissue Microarrays ◽  
Cell Types ◽  
Human Tissues ◽  
Expression Levels

CYP2S1 is a recently discovered dioxin-inducible member of the cytochrome P450 superfamily. It has been shown to be involved in the metabolism of some aromatic hydrocarbons as well as retinoic acid, suggesting a role in biotransformation of both exogenous and endogenous compounds. In this study, we used mRNA in situ hybridization and immunohistochemistry to investigate the cellular localization of CYP2S1 in various human tissues using tissue microarrays. High expression levels were observed mainly in epithelial cell types, especially in the epithelia frequently exposed to xenobiotics. In the respiratory tract, the expression was strong in nasal cavity, bronchi, and bronchioli, whereas it was low in the alveolar lining cells. Similarly, CYP2S1 was highly expressed in the epithelial cells throughout the gastrointestinal tract. Strong epithelial expression was also observed in uterine cervix, urinary bladder, and skin. In many exocrine glands (e.g., adrenal gland and pancreas), secretory epithelial cells showed moderate to strong expression levels. In the liver, the expression was low. CYP2S1 was highly expressed in epithelial cells that are major targets for carcinogen exposure and common progenitor cells to tumor development. Indeed, we found strong CYP2S1 expression in many tumors of epithelial origin.

scholarly journals Localization of Type 8 17β-hydroxysteroid Dehydrogenase mRNA in Mouse Tissues as Studied by In Situ Hybridization

2005 ◽  
Vol 53 (10) ◽  
pp. 1257-1271 ◽  
Author(s):  
Georges Pelletier ◽  
Van Luu-The ◽  
Songyun Li ◽  
Fernand Labrie
Keyword(s):  
In Situ Hybridization ◽  
Epithelial Cells ◽  
Stromal Cells ◽  
Cellular Localization ◽  
Pulmonary Arteries ◽  
Hydroxysteroid Dehydrogenase ◽  
Hybridization Signal ◽  
Mouse Tissues ◽  
Sites Of Action

The enzyme type 8 17β-hydroxysteroid dehydrogenase (17β-HSD) selectively catalyzes the conversion of estradiol (E2) to estrone (E1). To obtain detailed information on the sites of action of type 8 17β-HSD, we have studied the cellular localization of type 8 17β-HSD mRNA in mouse tissues using in situ hybridization. In the ovary, hybridization signal was detected in granulosa cells of growing follicles and luteal cells. In the uterus, type 8 17β-HSD mRNA was found in the epithelial (luminal and glandular) and stromal cells. In the female mammary gland, the enzyme mRNA was seen in ductal epithelial cells and stromal cells. In the testis, hybridization signal was observed in the seminiferous tubule. In the prostate, type 8 17β-HSD was detected in the epithelial cells of the acini and stromal cells. In the clitoral and preputial glands, labeling was detected in the epithelial cells of acini and small ducts. The three lobes of the pituitary gland were labeled. In the adrenal gland, hybridization signal was observed in the three zones of the cortex, the medulla being unlabeled. In the kidney, the enzyme mRNA was found to be expressed in the epithelial cells of proximal convoluted tubules. In the liver, all the hepatocytes exhibited a positive signal. In the lung, type 8 17β-HSD mRNA was detected in bronchial epithelial cells and walls of pulmonary arteries. The present data suggest that type 8 17β-HSD can exert its action to downregulate E2 levels in a large variety of tissues.

Developmental regulation of epithelial sodium channel subunit mRNA expression in rat colon and lung

1998 ◽  
Vol 275 (6) ◽  
pp. G1227-G1235 ◽  
Author(s):  
Shigeru Watanabe ◽  
Kazumichi Matsushita ◽  
John B. Stokes ◽  
Paul B. McCray
Keyword(s):  
In Situ Hybridization ◽  
Epithelial Cells ◽  
Mrna Expression ◽  
Developmental Regulation ◽  
Rat Colon ◽  
Rnase Protection ◽  
Quantitative Expression ◽  
Subunit Mrna ◽  
Lung And Kidney

Na+absorption via amiloride-sensitive Na+ channels is of critical importance in the transition between fetal and neonatal life in several tissues, including the colon, lung, and kidney. To characterize and contrast the mRNA expression of each of the three epithelial Na+ channel complex (ENaC) subunits, we conducted RNase protection assays (RPA) and in situ hybridization in colon and lung in fetal (17, 19, 20, and 21 days) and postnatal (1, 3, 9, 15, and 30 days) rats (r). In the colon the α-, β-, and γ-rENaC subunits showed quantitatively different but qualitatively similar expression. All three subunits gradually increased in abundance from fetal day 19 through day 30 of life. The amount of each subunit on day 30 was approximately three times the amount at day 1. In situ hybridization showed that each subunit was localized to the surface epithelial cells with minimal expression in the crypts. The lung showed a completely different pattern. In contrast to the colon, the total amount of α-rENaC mRNA (by RPA) in the lung increased dramatically from fetal day 19 to 21, whereas β- and γ-rENaC showed modest prenatal increases. The amounts of all three mRNAs fell after birth through day 9 (to about 75% of the day 1 value). On days 15 and 30 the amount of mRNA rose to approach the values on day 1. α-rENaC mRNA abundance always exceeded β- and γ-rENaC, and the quantitative expression was different for α- than for β- and γ-rENaC. In situ hybridization studies showed that all three subunits were expressed in epithelial cells of the bronchi, bronchioles, and alveoli and not in blood vessels. These studies show striking developmental heterogeneity in rENaC mRNA expression between lung and colon, probably reflecting different developmental regulatory mechanisms in these organs.

NHE2 and NHE3 are human and rabbit intestinal brush-border proteins

1996 ◽  
Vol 270 (1) ◽  
pp. G29-G41 ◽  
Author(s):  
W. A. Hoogerwerf ◽  
S. C. Tsao ◽  
O. Devuyst ◽  
S. A. Levine ◽  
C. H. Yun ◽  
...  
Keyword(s):  
Small Intestine ◽  
Epithelial Cells ◽  
Brush Border ◽  
Basolateral Membrane ◽  
Goblet Cells ◽  
Basolateral Membranes ◽  
Western Analysis ◽  
Rabbit Jejunum ◽  
Colon And Rectum ◽  
Descending Colon

Rabbit NHE2 and NHE3 are two epithelial isoform Na+/H+ exchangers (NHE), the messages for which are found predominantly and entirely, respectively, in renal, intestinal, and gastric mucosa. The current studies used Western analysis and immunohistochemistry to identify and characterize the apical vs. basolateral membrane distribution of NHE2 and NHE3 in intestinal epithelial cells. Based on Western analysis, NHE2 and NHE3 both are present in brush-border but not basolateral membranes of small intestine. Both NHE2 and NHE3 are 85-kDa proteins. Consistent with Western analysis, NHE2 and NHE3 are immunolocalired to the brush-border but not basolateral membranes of villus epithelial cells, but not goblet cells, in human jejunum and ileum and in surface epithelial cells in the ascending and descending colon and rectum. In addition, NHE2 and NHE3 are present in small amounts in the crypt cell brush border of human jejunum, ileum, ascending and descending colon, and rectum. In rabbit jejunum, ileum, and ascending colon, NHE2 and NHE3 are present in the brush border of epithelial and not goblet cells, again much more in the villus (small intestine)/ surface cells (colon) than the crypt. NHE2 but not NHE3 is present in the brush border of rabbit descending colon surface cells and in small amounts in crypt cells. NHE2 and NHE3 are both human and rabbit small intestinal and colonic epithelial cell brush-border Na+/H+ exchanger isoforms that colocalize in all intestinal segments except rabbit descending colon, which lacks NHE3.

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