The Villin1 Gene Promoter Drives Cre Recombinase Expression in Extraintestinal Tissues

Rutlin, Michael; Rastelli, Daniella; Kuo, Wei-Ting; Estep, Jason A.; Louis, Adriell; Riccomagno, Martin M.; Turner, Jerrold R.; Rao, Meenakshi

doi:10.1016/j.jcmgh.2020.05.009

Cited by 23 publications

(12 citation statements)

References 11 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To determine whether epithelial RET signaling was necessary for normal GI motility, we generated Vil1 Cre/1000 Ret flox/flox mice (hereafter RET EpiKO ) that would lack RET in the intestinal Epithelial RET signaling controls GI motility 10 epithelium but not in the ENS 32 . RET EpiKO mice were born at expected Mendelian ratios, had no overt deficits, and grew at the same pace as their littermate controls (Ret flox/flox mice; Supplementary Figure 6A-B).…”

Section: Ret Signaling In the Gut Epithelium Regulates Motility Selec...mentioning

confidence: 99%

RET Signaling Persists in the Adult Intestine and Stimulates Motility by Limiting PYY Release from Enteroendocrine Cells

Shepherd

Feinstein

Sabel

et al. 2022

Preprint

Self Cite

View full text Add to dashboard Cite

Background & Aims: RET receptor tyrosine kinase is necessary for enteric nervous system (ENS) development. Loss-of-function RET mutations cause Hirschsprung disease (HSCR), in which infants are born with aganglionic bowel. Despite surgical correction, HSCR patients often experience chronic defecatory dysfunction and enterocolitis, suggesting that RET is important after development. To test this hypothesis, we determined the location of postnatal RET and its significance in gastrointestinal (GI) motility. Methods: RetCFP/+ mice and human transcriptional profiling data were studied to identify the enteric neuronal and epithelial cells that express RET. To determine whether RET signaling in these cells regulates adult gut motility in vivo, genetic and pharmacologic approaches were used to disrupt RET in either all RET-expressing cells, a major subset of enteric neurons, or intestinal epithelial cells. Results: Distinct subsets of enteric neurons and enteroendocrine cells expressed RET in the adult intestine. RET disruption in the intestinal epithelium, rather than in enteric neurons, slowed GI motility selectively in adult male mice. This effect was phenocopied by RET kinase inhibition. Most RET+ epithelial cells were either enterochromaffin cells that release serotonin (5-HT) or L-cells that release peptide YY (PYY), both of which can alter motility. RET kinase inhibition exaggerated PYY release in a nutrient-dependent manner without altering 5-HT secretion. PYY receptor blockade fully rescued dysmotility in mice lacking epithelial RET. Conclusion: RET signaling normally limits nutrient-dependent PYY release from L-cells and this activity is necessary for normal intestinal motility in male mice. These effects could contribute to post-operative dysmotility in HSCR, which predominantly affects males, and uncovers a mechanism that could be targeted to treat post-prandial GI dysfunction.

show abstract

Section: Ret Signaling In the Gut Epithelium Regulates Motility Selec...mentioning

confidence: 99%

RET Signaling Persists in the Adult Intestine and Stimulates Motility by Limiting PYY Release from Enteroendocrine Cells

Shepherd

Feinstein

Sabel

et al. 2022

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…Cre recombinase expression in the intestinal epithelium is driven by the villin promotor, which allows for conditional tamoxifen-dependent Cre recombinase action to remove the Cnr1 gene from these cells, as described by el Marjou et al, [41]. When compared to other mouse lines that exhibit extra-intestinal expression of CRE recombinase, the Vil-CRE ERT2 mice used in our studies show selective expression in the intestinal epithelium with scattered expression in the testis [42]. Cnr1 tm1 .…”

Section: Transgenic Mouse Generationmentioning

confidence: 75%

Cannabinoid CB1 Receptors in the Intestinal Epithelium Are Required for Acute Western-Diet Preferences in Mice

et al. 2020

View full text Add to dashboard Cite

The endocannabinoid system plays an important role in the intake of palatable food. For example, endocannabinoid signaling in the upper small-intestinal epithelium is increased (i) in rats after tasting dietary fats, which promotes intake of fats, and (ii) in a mouse model of diet-induced obesity, which promotes overeating via impaired nutrient-induced gut–brain satiation signaling. We now utilized a combination of genetic, pharmacological, and behavioral approaches to identify roles for cannabinoid CB1Rs in upper small-intestinal epithelium in preferences for a western-style diet (WD, high-fat/sucrose) versus a standard rodent diet (SD, low-fat/no sucrose). Mice were maintained on SD in automated feeding chambers. During testing, mice were given simultaneous access to SD and WD, and intakes were recorded. Mice displayed large preferences for the WD, which were inhibited by systemic pretreatment with the cannabinoid CB1R antagonist/inverse agonist, AM251, for up to 3 h. We next used our novel intestinal epithelium-specific conditional cannabinoid CB1R-deficient mice (IntCB1−/−) to investigate if intestinal CB1Rs are necessary for WD preferences. Similar to AM251 treatment, preferences for WD were largely absent in IntCB1−/− mice when compared to control mice for up to 6 h. Together, these data suggest that CB1Rs in the murine intestinal epithelium are required for acute WD preferences.

show abstract

“…By using breeders that were heterozygous for the Cre allele, we generated littermate controls (IEC WT ) that were co-housed with their IEC ΔMHCII littermates during all experiments, to prevent any potential differences in the microbiota. In addition, as there have been reports that the Villin-Cre strain can be ‘leaky’, resulting in off-target deletion of the floxed alleles 28 , we validated each individual IEC ΔMHCII mouse by flow cytometry to verify selective ablation of MHC II in IECs, with maintenance of MHC II expression in haematopoietic lineages (Figure S2A and B) .…”

Section: Resultsmentioning

confidence: 99%

“…Although many factors could contribute to these discrepancies, high mortality following infection with a pathogen that normally elicits a mild, self-limiting disease suggests fundamental biological differences. Some Villin-Cre transgenic lines can exhibit a low level of Cre recombinase activity in extraintestinal tissues which can lead to germline deletion of the floxed alleles 28 . Indeed, we observed this in multiple strains we generated using Villin-Cre mice, including the MHC II I-Ab 'floxed' strain used in this study.…”

Section: Discussionmentioning

confidence: 99%

MHC class II antigen presentation by intestinal epithelial cells fine-tunes bacteria-reactive CD4 T cell responses

Heuberger

Janney

Ilott

et al. 2023

Preprint

View full text Add to dashboard Cite

Although intestinal epithelial cells (IECs) can express major histocompatibility complex class II (MHC II), especially during intestinal inflammation, it remains unclear if antigen presentation by IECs favours pro- or anti-inflammatory CD4+ T cell responses. Using selective gene ablation of MHC II in IECs and IEC organoid cultures, we assessed the impact of MHC II expression by IECs on CD4+ T cell responses and disease outcomes in response to enteric bacterial pathogens. We found that intestinal bacterial infections elicit inflammatory cues that greatly increase expression of MHC II processing and presentation molecules in colonic IECs. Whilst IEC MHC II expression had little impact on disease severity following Citrobacter rodentium or Helicobacter hepaticus infection, using a colonic IEC organoid-CD4+ T cell co-culture system, we demonstrate that IECs can activate antigen-specific CD4+ T cells in an MHC II-dependent manner, modulating both regulatory and effector Th cell subsets. Furthermore, we assessed adoptively transferred H. hepaticus-specific CD4+ T cells during intestinal inflammation in vivo and report that IEC MHC II expression dampens pro-inflammatory effector Th cells. Our findings indicate that IECs can function as non-conventional antigen presenting cells and that IEC MHC II expression fine-tunes local effector CD4+ T cell responses during intestinal inflammation.

show abstract

The Villin1 Gene Promoter Drives Cre Recombinase Expression in Extraintestinal Tissues

Cited by 23 publications

References 11 publications

RET Signaling Persists in the Adult Intestine and Stimulates Motility by Limiting PYY Release from Enteroendocrine Cells

RET Signaling Persists in the Adult Intestine and Stimulates Motility by Limiting PYY Release from Enteroendocrine Cells

Cannabinoid CB1 Receptors in the Intestinal Epithelium Are Required for Acute Western-Diet Preferences in Mice

MHC class II antigen presentation by intestinal epithelial cells fine-tunes bacteria-reactive CD4 T cell responses

Contact Info

Product

Resources

About