Dataset Information

Tissue and cellular expression patterns of porcine CFTR: similarities to and differences from human CFTR.

ABSTRACT: Emerging porcine models of cystic fibrosis (CF) are expected to mimic the human disease more closely than current mouse models do. However, little is known of the tissue and cellular expression patterns of the porcine CF transmembrane conductance regulator (pCFTR) and possible differences from human CFTR (hCFTR). Here, the expression pattern of pCFTR was systematically established on the mRNA and protein levels. Using specific anti-pCFTR antibodies, the majority of the protein was immunohistochemically detected on paraffin-embedded sections and on cryostate sections in the apical cytosol of intestinal crypt epithelial cells, nasal, tracheal, and bronchial epithelial cells, and other select, mostly glandular epithelial cells. Confocal laser scanning microscopy with co-localization of the Golgi marker 58K localized the protein in the cytosol between the Golgi apparatus and the apical cell membrane with occasional punctate or diffuse staining of the apical membrane. The tissue and cellular distribution patterns were confirmed by RT-PCR from whole tissue lysates or select cells after laser capture microdissection. Thus, expression of pCFTR was found to largely resemble that of hCFTR except for the kidney, brain, and cutaneous glands, which lack expression in pigs. Species-specific differences between pCFTR and hCFTR may become relevant for future interpretations of the CF phenotype in pig models.

SUBMITTER: Plog S

PROVIDER: S-EPMC2924795 | biostudies-literature | 2010 Sep

REPOSITORIES: biostudies-literature

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Tissue and cellular expression patterns of porcine CFTR: similarities to and differences from human CFTR.

Plog Stephanie S Mundhenk Lars L Bothe Melanie K MK Klymiuk Nikolai N Gruber Achim D AD

The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society 20100524 9

Emerging porcine models of cystic fibrosis (CF) are expected to mimic the human disease more closely than current mouse models do. However, little is known of the tissue and cellular expression patterns of the porcine CF transmembrane conductance regulator (pCFTR) and possible differences from human CFTR (hCFTR). Here, the expression pattern of pCFTR was systematically established on the mRNA and protein levels. Using specific anti-pCFTR antibodies, the majority of the protein was immunohistoche ...[more]

PMID: 20498480

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Project description:AIM:The present study was aimed to investigate the developmental patterns of growth hormone receptor (GHr) and somatostatin (SS) mRNA expression in porcine gastric tissue and its relationship with gastric growth and gastric functional development. METHODS:Erhualian and Large White boars were selected randomly and sampled at birth (D0), 3, 20, 30, 90, 120 and 180 days of age respectively, meanwhile the bodyweight and gastric weight were recorded. The single tube semi-quantitative RT-PCR was applied in this experiment to investigate the developmental patterns of gastric GHr and SS mRNA expression, the correlations between the patterns of mRNA expression and the relative gastric weight (ratio of gastric weight to bodyweight) and the pepsin contents in gastric mucous membrane were analyzed. In order to further elucidate the effect of GH on gastric function, the primary cultures of gastric fundic mucosal cells were treated with 2 ng/ml, 20 ng/ml and 200 ng/ml of rpGH for 18 hrs respectively, and the pepsin contents in culture medium were measured. RESULTS:The expression of GHr mRNA was high at birth, followed by a significant decrease at 3 days of age (D3) in both breeds. In Large White boars, the expression of GHr mRNA reached a peak at D90 and remained a plateau afterward. In Erhualian pigs, however, a slight yet significant increase occurred at D30 reaching a level that was kept constant thereafter. From birth to D30, the expression of GHr mRNA in gastric tissue was higher in Erhualian boars than that in Large White boars, but from D90 to D180, the higher expression of GHr mRNA was found in Large White boars. The gastric GHr mRNA expression was significantly correlated with the relative gastric weight (r=0.541, P<0.05) and pepsin content in gastric mucosa (r=0.625, P<0.05) respectively. The gastric SS mRNA expression was high at birth (Erhualian 1.59, Large White 0.80), but dropped significantly at D3 (Erhualian 0.95, Large White 0.19, P<0.05), a stepwise increase was followed thereafter until a peak at D30 (Erhualian 1.71, Large White 0.95) In general, Erhualian pigs expressed higher levels of SS mRNA in gastric tissue as compared with Large White pigs at the same age (P<0.05). No significant correlations between SS mRNA and relative gastric weight or pepsin content were found. In vitro, 2 ng/ml of rpGH elicited significant increase in pepsin secretion from primary cultures of gastric mucosal cells (P<0.05), and no responses were observed at 20 ng/ml and 200 ng/ml. CONCLUSION:The results suggested that: 1, GHr and SS mRNA in porcine stomach are expressed according to strain specific developmental patterns; 2, GH acts directly at the gastric tissue and regulates the structural and functional growth of stomach.

Project description:BackgroundAccumulation of visceral adipose tissue (VAT) is clearly associated with an increased risk of obesity-related diseases and all-cause mortality, whereas gluteal subcutaneous fat accumulation (g-SAT) is associated with a lower risk. The relative contribution, in term of cardiovascular risk, of abdominal subcutaneous adipose tissue (a-SAT) is still controversial with studies showing both a detrimental effect and a protective role. Animal and in vitro studies demonstrated that adipocytes from visceral and subcutaneous depots have distinct morphological, metabolic and functional characteristics. These regional differences have a key role in the pathogenesis of obesity-related diseases. There is recent evidence that differentiation between upper-body and lower-body adipose tissues might be under control of site-specific sets of developmental genes, such as Homebox (HOX) genes, a group of related genes that control the body plan of an embryo along the anterior-posterior axis. However, the possible heterogeneity between different subcutaneous regions has not been extensively investigated. Here we studied global mRNA expression in g-SAT and a-SAT with a microarray approach. RNA was isolated from g-SAT and a-SAT biopsy, from eight healthy subjects, and hybridized on RNA microarray chips in order to detect regional differences in gene expression.ResultsA total of 131 genes are significantly and differently (>1.5 fold change, p < 0.05) expressed in a-SAT and g-SAT. Expression profiling reveals significant differences in expression of several HOX genes. Interestingly, two molecular signature of visceral adipocyte lineage, homebox genes HOXA5 and NR2F1, are up-regulated in a-SAT versus g-SAT by a 2.5 fold change.ConclusionsOur study shows that g-SAT and a-SAT have distinct expression profiles. The finding of a different expression of HOX genes, fundamental during the embryo development, suggests an early regional differentiation of subcutaneous adipose depots. Moreover, the higher expression of HOXA5 and NR2F1, two molecular signatures of visceral adipocytes, in a-SAT suggests that this subcutaneous adipose depot could be more similar to VAT than g-SAT. Our data suggest that we should look at SAT as composed of distinct depots with possibly different impact in obesity associated metabolic complications.

Dataset Information

Tissue and cellular expression patterns of porcine CFTR: similarities to and differences from human CFTR.

Publications

Tissue and cellular expression patterns of porcine CFTR: similarities to and differences from human CFTR.

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