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

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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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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Goblet-cell-specific transcription of mouse intestinal trefoil factor gene results from collaboration of complex series of positive and negative regulatory elements

Biochemical Journal ◽

10.1042/bj3410461 ◽

1999 ◽

Vol 341 (2) ◽

pp. 461-472 ◽

Cited By ~ 14

Author(s):

Hiroshi ITOH ◽

Nagamu INOUE ◽

Daniel K. PODOLSKY

Keyword(s):

Cell Lines ◽

Goblet Cell ◽

Regulatory Element ◽

Regulatory Region ◽

Goblet Cells ◽

Regulatory Elements ◽

Intestinal Cell ◽

Gene Promoters ◽

Trefoil Factor ◽

Intestinal Trefoil Factor

Intestinal trefoil factor (ITF) is expressed selectively in intestinal goblet cells. Previous studies of the rat ITF gene identified one cis-regulatory element, designated the goblet-cell-response element (GCRE), present in the proximal region of the promoter. To identify additional cis-regulatory elements responsible for goblet-cell-specific expression, a DNA fragment containing 6353 bp of the 5′-flanking region of the mouse ITF gene was cloned and its promoter activity was examined extensively. In human and murine intestinal-derived cell lines (LS174T and CMT-93), the luciferase activities of a 6.3-kb construct were 5- and 2-fold greater than the smaller 1.8-kb construct, respectively. In contrast, the activity in non-intestinal cell lines (HepG2 and HeLa) was 2-4-fold lower than the smaller construct. In the region downstream from the 1.8-kb position, strong luciferase activities in LS174T and HepG2 cells were observed using a 201-bp construct. Interestingly, increased activity was almost completely suppressed in cells transfected with a 391-bp construct. Detailed analyses of this region revealed the existence of a 11-bp positive regulatory element (-181 to -170; ACCTCTTCCTG) and a 9-bp negative regulatory element (-208 to -200; ATTGACAGA) in addition to the GCRE. All three elements were well conserved among human, rat and mouse ITF gene promoters. In addition, a mutant 1.8-kb construct in which the negative regulatory region was deleted yielded the same approximate luciferase activity as a 6.3-kb construct, suggesting binding of a goblet-cell-specific silencer inhibitor (SI) between -6.3 and -1.8 kb. The SI present in goblet cells may block the silencers' binding to the pre-initiation complex and allow increased transcriptional activity driven by specific and non-specific enhancers. High-level expression of the mouse ITF gene specifically in intestinal goblet cells may be achieved through the combined effects of these regulatory elements.

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A silencer inhibitor confers specific expression of intestinal trefoil factor in gobletlike cell lines

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.2001.280.6.g1114 ◽

2001 ◽

Vol 280 (6) ◽

pp. G1114-G1123 ◽

Cited By ~ 15

Author(s):

Dai Iwakiri ◽

Daniel K. Podolsky

Keyword(s):

Cell Lines ◽

Goblet Cell ◽

Regulatory Element ◽

Goblet Cells ◽

Transient Transfection ◽

Trefoil Factor ◽

Intestinal Trefoil Factor ◽

Specific Expression ◽

Silencer Element

Intestinal trefoil factor (ITF) is selectively expressed in intestinal goblet cells. Previous studies identified cis-regulatory elements in the proximal promoter of ITF, but these were insufficient to recapitulate the exquisite tissue- and cell-specific expression of native ITF in vivo. Preliminary studies suggested that goblet cell-specific expression of murine ITF requires elements far upstream that include a silencer element that effectively prevents ITF expression in non-goblet cells. Transient transfection studies using native or mutant ITF 5′-flanking sequences identified a region that restores expression in goblet cells. This element, designated goblet cell silencer inhibitor (GCSI) element, enables human and murine goblet cell-like cell lines to override the silencing effect of more proximal elements. The GCSI has no intrinsic enhancer activity and regulates expression only when the silencer element is present. Ligation of GCSI and silencer elements to sucrase-isomaltase conferred goblet cell-specific expression. Goblet cells but not non-goblet cells possess a nuclear protein that binds to the GCSI regulatory element (GCSI binding protein; GCSI-BP). Both transient transfection and gel mobility shift assay studies localize the GCSI and GCSI-BP to −2216 to −2204. We conclude that goblet cell-specific transcription of ITF in vivo depends on a regulatory element designated GCSI.

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Keratinocyte growth factor regulates expression of the intestinal trefoil factor gene through a cis-regulatory element regulating goblet cell-specific transcription

Gastroenterology ◽

10.1016/s0016-5085(00)83857-3 ◽

2000 ◽

Vol 118 (4) ◽

pp. A436

Author(s):

Dai Iwakiri ◽

Daniel K. Podolsky

Keyword(s):

Growth Factor ◽

Goblet Cell ◽

Regulatory Element ◽

Keratinocyte Growth Factor ◽

Trefoil Factor ◽

Intestinal Trefoil Factor ◽

Factor Gene

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Activation of intestinal tuft cell-expressed Sucnr1 triggers type 2 immunity in the mouse small intestine

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1720758115 ◽

2018 ◽

Vol 115 (21) ◽

pp. 5552-5557 ◽

Cited By ~ 61

Author(s):

Weiwei Lei ◽

Wenwen Ren ◽

Makoto Ohmoto ◽

Joseph F. Urban ◽

Ichiro Matsumoto ◽

...

Keyword(s):

Goblet Cell ◽

Mucosal Immunity ◽

Cell Activation ◽

Nippostrongylus Brasiliensis ◽

Infectious Agents ◽

Cell Hyperplasia ◽

Type 2 Immunity ◽

Tuft Cell ◽

Tuft Cells

The hallmark features of type 2 mucosal immunity include intestinal tuft and goblet cell expansion initiated by tuft cell activation. How infectious agents that induce type 2 mucosal immunity are detected by tuft cells is unknown. Published microarray analysis suggested that succinate receptor 1 (Sucnr1) is specifically expressed in tuft cells. Thus, we hypothesized that the succinate–Sucnr1 axis may be utilized by tuft cells to detect certain infectious agents. Here we confirmed that Sucnr1 is specifically expressed in intestinal tuft cells but not in other types of intestinal epithelial cells, and demonstrated that dietary succinate induces tuft and goblet cell hyperplasia via Sucnr1 and the tuft cell-expressed chemosensory signaling elements gustducin and Trpm5. Conventional mice with a genetic Sucnr1 deficiency (Sucnr1−/−) showed diminished immune responses to treatment with polyethylene glycol and streptomycin, which are known to enhance microbiota-derived succinate, but responded normally to inoculation with the parasitic worm Nippostrongylus brasiliensis that also produces succinate. Thus, Sucnr1 is required for microbiota-induced but not for a generalized worm-induced type 2 immunity.

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Identification of a Goblet Cell-specific Enhancer Element in the Rat Intestinal Trefoil Factor Gene Promoter Bound by a Goblet Cell Nuclear Protein

Journal of Biological Chemistry ◽

10.1074/jbc.273.5.3060 ◽

1998 ◽

Vol 273 (5) ◽

pp. 3060-3067 ◽

Cited By ~ 31

Author(s):

Haruhiko Ogata ◽

Nagamu Inoue ◽

Daniel K. Podolsky

Keyword(s):

Goblet Cell ◽

Nuclear Protein ◽

Gene Promoter ◽

Trefoil Factor ◽

Intestinal Trefoil Factor ◽

Enhancer Element ◽

Factor Gene

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Immunohistochemical Analyses of MUC5AC Mucin Expression in Sinus Mucosa of Children with Sinusitis and Controls

Annals of Otology Rhinology & Laryngology ◽

10.1177/000348940511401212 ◽

2005 ◽

Vol 114 (12) ◽

pp. 958-965 ◽

Cited By ~ 10

Author(s):

Maria T. Peña ◽

Pawandeep K. Aujla ◽

Kantilal M. Patel ◽

George H. Zalzal ◽

Mary C. Rose

Keyword(s):

Goblet Cell ◽

Chronic Sinusitis ◽

Goblet Cells ◽

Surface Epithelium ◽

Cell Hyperplasia ◽

Goblet Cell Hyperplasia ◽

Mucin Expression ◽

Glandular Cells ◽

Sinus Mucosa ◽

Immunohistochemical Analyses

Objectives: The purpose of this study was to analyze MUC5AC protein expression in sinus mucosal specimens of children with and without chronic sinusitis. Methods: Morphometric, histologic, and immunohistochemical analyses were carried out on sinus mucosa of 7 children with chronic sinusitis and 6 children without sinusitis. Results: MUC5AC protein was expressed in a subset of goblet cells in the surface epithelium, but not in the submucosal glands in either pediatric population. The number of goblet cells that expressed MUC5AC mucin was not significantly different in patients with and without chronic sinusitis. All specimens had similar numbers of goblet cells in the surface epithelium. Conclusions: The data demonstrate that neither goblet cell hyperplasia nor increased MUC5AC expression occurs in the sinus mucosa of children with chronic sinusitis. This suggests that in contrast to asthma, in which goblet cell hyperplasia is present in the lower respiratory tract, mucus hypersecretion in pediatric chronic sinusitis may involve other secretory cells, eg, submucosal glandular cells, and mucins secreted by these glandular cells.

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