Localization of intestinal trefoil-factor mRNA in rat stomach and intestine by hybridization in situ

A cDNA encoding rat intestinal trefoil factor (rITF) was prepared by reverse transcription and PCR amplification. The sequence obtained was well conserved with that of other trefoil peptides. An antisense riboprobe produced from the clone was used to localize the sites of ITF expression in the rat gastrointestinal tract using hybridization in situ. We found rITF mRNA in goblet cells in the small intestine and colon; a gradient of signal strength greatest near the crypt base was sometimes present. We found no evidence for rITF expression in Brunner's glands, the pancreas, or most regions of the gastric mucosa. Surprisingly, strong signals for rITF mRNA were detected in a region of stomach at the junction of the squamous fore-stomach with the glandular gastric mucosa. This region, which may correspond to the cardiac region, formed part of a larger area of cells staining positive for acid mucins. We hypothesize that concerted expression occurs of particular trefoil peptides with specific mucins, and that this organization reflects a functional relationship between mucins and trefoil peptides.

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cDNA cloning of rat pS2 peptide and expression of trefoil peptides in acetic acid-induced colitis

Biochemical Journal ◽

10.1042/bj3180939 ◽

1996 ◽

Vol 318 (3) ◽

pp. 939-944 ◽

Cited By ~ 28

Author(s):

Hiroshi ITOH ◽

Masaki TOMITA ◽

Hirofumi UCHINO ◽

Takahiko KOBAYASHI ◽

Hiroaki KATAOKA ◽

...

Keyword(s):

Acetic Acid ◽

Acute Phase ◽

Recovery Phase ◽

Intestinal Epithelial ◽

Trefoil Factor ◽

Intestinal Trefoil Factor ◽

Reverse Transcription Pcr ◽

Trefoil Peptides ◽

Rapid Amplification ◽

Normal Controls

By using a combination of the methods of reverse transcription-PCR and rapid amplification of cDNA ends, a cDNA for rat pS2 peptide (rpS2) was successfully cloned and sequenced from rat stomach. By RNA blot analysis, the gene was shown to be expressed abundantly in the stomach and only faintly in the duodenum, but not in other tissues including the distal small and large intestines. rpS2 expression was also examined in the rectum during the course of acetic acid-induced colitis; rpS2 mRNA was detected during the acute phase of colitis but not in normal controls or during the recovery phase. On the other hand, expression of rat intestinal trefoil factor (rITF) was down-regulated during the acute phase of colitis and then up-regulated during the recovery phase, whereas rat spasmolytic peptide was not detectable throughout the course of the induced colitis. These results indicate that the patterns and timing of the expression of these trefoil peptides are different from each other. rpS2 may play an important role in the proliferation of intestinal epithelial cells during the acute phase of mucosal ulceration, whereas rITF may be involved in differentiation of the cells, particularly to form goblet cells, during the recovery phase.

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Histochemical Assessment of Mucin-Secreting Cells from the Gastric Mucosa of Guinea Pigs

Bulletin of University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca Veterinary Medicine ◽

10.15835/buasvmcn-vm:2020.0032 ◽

2020 ◽

Vol 77 (2) ◽

pp. 115

Author(s):

Adriana CHENDE ◽

Vasile RUS ◽

Cristian MARTONOS ◽

Dalma PIVARIU ◽

Aurel DAMIAN ◽

...

Keyword(s):

Gastric Mucosa ◽

Guinea Pig ◽

Alcian Blue ◽

Guinea Pigs ◽

Classical Method ◽

Cardiac Region ◽

Pas Reaction ◽

Acid Mucins ◽

Blue Reaction ◽

Number Of Cells

Stomach fragments from 3 guinea pigs were collected from the three regions: cardiac, fundic and pyloric, for histochemical investigations. The anatomical segments were processed by the classical method of inclusion in paraffin and the histological sections were stained with PAS reaction for highlighting the neutral mucins and the Alcian blue method for acid mucins. All the surface cells of the gastric mucosa and in the crypts were positive on the PAS reaction and negative in the case of the alcian blue reaction. This demonstrates that cells on the surface and in the crypts synthetize neutral mucins. In the case of the cardiac region glands, only a small number of cells were positive on the two histochemical reactions, which shows that the cardiac glands in guinea pig synthesize a very small amount of neutral and acidic mucins. There are no positive cells in neither reaction used in the fundic glands case, which shows that these glands do not synthesize mucins, neither neutral nor acidic content. The glands located in the pyloric region have cells in the deep half of the wall which were positive on both histochemical reactions, which shows that they synthesize both neutral mucins and acidic mucins.

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Characterisation of the single copy trefoil peptides intestinal trefoil factor and pS2 and their ability to form covalent dimers

FEBS Letters ◽

10.1016/0014-5793(94)01297-e ◽

1995 ◽

Vol 357 (1) ◽

pp. 50-54 ◽

Cited By ~ 39

Author(s):

Rebecca Chinery ◽

Paul A. Bates ◽

Amitabha De ◽

Paul S. Freemont

Keyword(s):

Single Copy ◽

Trefoil Factor ◽

Intestinal Trefoil Factor ◽

Trefoil Peptides

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The trefoil peptides spasmolytic polypeptide and intestinal trefoil factor are major secretory products of the rat gut

Peptides ◽

10.1016/0196-9781(95)00070-z ◽

1995 ◽

Vol 16 (6) ◽

pp. 1001-1005 ◽

Cited By ~ 35

Author(s):

D.R. Taupin ◽

K.C. Pang ◽

S.P. Green ◽

A.S. Giraud

Keyword(s):

Trefoil Factor ◽

Intestinal Trefoil Factor ◽

Trefoil Peptides ◽

Spasmolytic Polypeptide ◽

Secretory Products ◽

Rat Gut

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Visualizing miRNAs in adult skeletal muscle by hybridization in situ

Protocol Exchange ◽

10.1038/nprot.2006.136 ◽

2006 ◽

Author(s):

Anna Polesskaya

Keyword(s):

Skeletal Muscle ◽

Adult Skeletal Muscle ◽

Hybridization In Situ

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The Alcohol Dehydrogenase Gene Is Nested in the outspread Locus of Drosophila melanogaster

Genetics ◽

10.1093/genetics/143.2.897 ◽

1996 ◽

Vol 143 (2) ◽

pp. 897-911 ◽

Cited By ~ 1

Author(s):

S McNabb ◽

S Greig ◽

T Davis

Keyword(s):

Drosophila Melanogaster ◽

Alcohol Dehydrogenase ◽

Pcr Amplification ◽

Transcription Unit ◽

Adjacent Gene ◽

Dehydrogenase Gene ◽

Chromosomal Breakpoints ◽

Splicing Patterns ◽

Somatic Musculature

Abstract This report describes the structure and expression of the outspread (osp) gene of Drosophila melanogaster. Previous work showed that chromosomal breakpoints associated with mutations of the osp locus map to both sides of the alcohol dehydrogenase gene (Adh), suggesting that Adh and the adjacent gene Adh' are nested in osp. We extended a chromosomal walk and mapped additional osp mutations to define the maximum molecular limit of osp as 119 kb. We identified a 6-kb transcript that hybridizes to osp region DNA and is altered or absent in osp mutants. Accumulation of this RNA peaks during embryonic and pupal periods. The osp cDNAs comprise two distinct classes based on alternative splicing patterns. The 5′ end of the longest cDNA was extended by PCR amplification. When hybridized to the osp walk, the 5′ extension verifies that Adh and Adh' are nested in osp and shows that osp has a transcription unit of ≥74 kb. In situ hybridization shows that osp is expressed both maternally and zygotically. In the ovary, osp is transcribed in nurse cells and localized in the oocyte. In embryos, expression is most abundant in the developing visceral and somatic musculature.

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Germline Transformation of Drosophila virilis With the Transposable Element mariner

Genetics ◽

10.1093/genetics/143.1.365 ◽

1996 ◽

Vol 143 (1) ◽

pp. 365-374 ◽

Cited By ~ 1

Author(s):

Allan R Lohe ◽

Daniel L Hartl

Keyword(s):

Transposable Element ◽

Common Ancestor ◽

Pcr Amplification ◽

Drosophila Virilis ◽

Cell Region ◽

Diverse Species ◽

Drosophila Mauritiana ◽

Characteristic 2 ◽

In Situ Hybridizations

Abstract An important goal in molecular genetics has been to identify a transposable element that might serve as an efficient transformation vector in diverse species of insects. The transposable element mariner occurs naturally in a wide variety of insects. Although virtually all mariner elements are nonfunctional, the Mosl element isolated from Drosophila mauritiana is functional. Mosl was injected into the pole-cell region of embryos of D. virilis, which last shared a common ancestor with D. mauritiana 40 million years ago. Mosl PCR fragments were detected in several pools of DNA from progeny of injected animals, and backcross lines were established. Because Go lines were pooled, possibly only one transformation event was actually obtained, yielding a minimum frequency of 4%. Mosl segregated in a Mendelian fashion, demonstrating chromosomal integration. The copy number increased by spontaneous mobilization. In situ hybridization confirmed multiple polymorphic locations of Mosl. Integration results in a characteristic 2-bp TA duplication. One Mosl element integrated into a tandem array of 370-bp repeats. Some copies may have integrated into heterochromatin, as evidenced by their ability to support PCR amplification despite absence of a signal in Southern and in situ hybridizations.

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Molecular cloning of mouse intestinal trefoil factor and its expression during goblet cell changes

Biochemical Journal ◽

10.1042/bj3110293 ◽

1995 ◽

Vol 311 (1) ◽

pp. 293-297 ◽

Cited By ~ 36

Author(s):

M Tomita ◽

H Itoh ◽

N Ishikawa ◽

A Higa ◽

H Ide ◽

...

Keyword(s):

Goblet Cell ◽

Goblet Cells ◽

Recovery Phase ◽

Nippostrongylus Brasiliensis ◽

Trefoil Factor ◽

Intestinal Trefoil Factor ◽

Cell Hyperplasia ◽

Reverse Transcription Pcr ◽

Rapid Amplification ◽

Mitf Expression

A cDNA encoding mouse intestinal trefoil factor (mITF) was successfully cloned and sequenced from the small intestine of C57BL/6 mouse by using the combination of reverse transcription-PCR and rapid amplification of cDNA ends methods. The gene was, similar to rat and human ITFs, mainly expressed in the small and large intestine. The mITF expression was up-regulated during the recovery phase after depletion of goblet cells in acetic acid-induced colitis. On the other hand, the expression in the jejunum was not altered, while goblet cell hyperplasia was induced by Nippostrongylus brasiliensis infection. These results suggest that the mITF expression did not simply correlate with the number of goblet cells. The mITF may play an important role in the maintenance and repair of mucosal function of the rectum. Additionally, the mITF in the jejunum may play a role in alteration of the physicochemical nature of goblet cell mucins, thereby affecting the establishment of intestinal helminths.

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