Mouse Insulin Cells Expressing an Inducible RIPCre Transgene Are Functionally Impaired

Teitelman, Gladys; Kedees, Mamdouh H.

doi:10.1074/jbc.m114.615484

Cited by 10 publications

(10 citation statements)

References 29 publications

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“…Given concerns that Cre-recombinase expression or activity may lead to artifactual phenotypes ( 2 , 4 ), we next evaluated whole-body glucose homeostasis and in vivo insulin secretion in tamoxifen-treated MIP-CreERT mice on chow and HFHS diets. Compared with wild-type (WT) littermates, chow-fed MIP-CreERT mice showed no significant differences in weight, oral glucose tolerance, insulin tolerance, or fasting or refed glucose ( Fig.…”

Section: Resultsmentioning

confidence: 99%

“…This difference may be due to transgene-specific differences in the level of hGH or secretion or other factors independent of hGH and serotonin action. Thus, it remains unclear whether ectopic hGH in β-cells is the underlying cause of cellular and metabolic abnormalities (including impaired β-cell function) reported for other Cre-recombinase transgenic lines ( 2 – 4 ).…”

Section: Discussionmentioning

confidence: 99%

“…In this regard, β-cells appear particularly sensitive. A few widely used transgenic mouse lines created using the promoters of pancreatic and duodenal homeobox 1 (Pdx1) or the rat insulin promoter (RIP) are reported to have significant transgene-mediated effects on insulin secretion and glucose tolerance ( 2 – 4 ). In addition, ectopic Cre-recombinase expression in the central nervous system (CNS) may cause non–β-cell related physiological effects ( 5 , 6 ).…”

Section: Introductionmentioning

confidence: 99%

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Phenotypic Characterization of MIP-CreERT1Lphi Mice With Transgene-Driven Islet Expression of Human Growth Hormone

et al. 2015

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There is growing concern over confounding artifacts associated with β-cell–specific Cre-recombinase transgenic models, raising questions about their general usefulness in research. The inducible β-cell–specific transgenic (MIP-CreERT1Lphi) mouse was designed to circumvent many of these issues, and we investigated whether this tool effectively addressed concerns of ectopic expression and disruption of glucose metabolism. Recombinase activity was absent from the central nervous system using a reporter line and high-resolution microscopy. Despite increased pancreatic insulin content, MIP-CreERT mice on a chow diet exhibited normal ambient glycemia, glucose tolerance and insulin sensitivity, and appropriate insulin secretion in response to glucose in vivo and in vitro. However, MIP-CreERT mice on different genetic backgrounds were protected from high-fat/ streptozotocin (STZ)-induced hyperglycemia that was accompanied by increased insulin content and islet density. Ectopic human growth hormone (hGH) was highly expressed in MIP-CreERT islets independent of tamoxifen administration. Circulating insulin levels remained similar to wild-type controls, whereas STZ-associated increases in α-cell number and serum glucagon were significantly blunted in MIP-CreERT1Lphi mice, possibly due to paracrine effects of hGH-induced serotonin expression. These studies reveal important new insight into the strengths and limitations of the MIP-CreERT mouse line for β-cell research.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Phenotypic Characterization of MIP-CreERT1Lphi Mice With Transgene-Driven Islet Expression of Human Growth Hormone

et al. 2015

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“…The mammalian DNA-damaging effect of Cre probably depends on its expression level and the cell type where Cre is expressed (3). One well-studied example are mice that express Cre under the rat insulin promoter (rip) and become insulin resistant in the absence of any loxP-targeted gene deletion (10,11). Thus, it was proposed that a large body of published work using rip-cre + mice and claiming effects of several genes on glucose intolerance/diabetes, in which rip-cre + control mice were not included, might be incorrect (10).…”

mentioning

confidence: 99%

lck-Driven Cre Expression Alters T Cell Development in the Thymus and the Frequencies and Functions of Peripheral T Cell Subsets

Carow

Gao

Coquet

et al. 2016

The Journal of Immunology

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Conditional gene targeting using the bacteriophage-derived Cre recombinase is widely applied for functional gene studies in mice. Mice transgenic for Cre under the control of the lck gene promoter are used to study the role of loxP-targeted genes in T cell development and function. In this article, we show a striking 65% reduction in cellularity, preferential development of γδ versus αβ T cells, and increased expression of IL-7R in the thymus of mice expressing Cre under the proximal lck promoter (lck-cre+ mice). The transition from CD4/CD8 double-negative to double-positive cells was blocked, and lck-cre+ double-positive cells were more prone to apoptosis and showed higher levels of Cre expression. Importantly, numbers of naive T cells were reduced in spleens and lymph nodes of lck-cre+ mice. In contrast, frequencies of γδ T cells, CD44+CD62L− effector T cells, and Foxp3+ regulatory T cells were elevated, as was the frequency of IFN-γ–secreting CD4+ and CD8+ T cells. A literature survey of 332 articles that used lck-cre+ mice for deletion of floxed genes indicated that results are statistically influenced by the control used (lck-cre+ or lck-cre−), more frequently resembling the lck-cre+ phenotype described in this article if lck-cre− controls were used. Altogether, care should be taken when interpreting published results and to properly control targeted gene deletions using the lck-cre+ strain.

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“…Some studies indicated that tamoxifen and RU486 interfere with reproductive and heart physiology, which could affect interpretation of acquired phenotypes (39,40). Moreover, emerging evidence indicates that constitutive expression of full-length functional Cre or Cre ER protein exhibits serious metabolic side effects, such as Nestin-Cre on pituitary functions as well as Rip-Cre or Cre ER on β-cell functions (3,41). In contrast, PTC promotes NMD, a conserved pathway that destroys the majority of nsCre transcripts, thereby leaving only a few mutant mRNAs that are translated into truncated peptides (13).…”

Section: Discussionmentioning

confidence: 99%

New inducible genetic method reveals critical roles of GABA in the control of feeding and metabolism

Meng

Han

Srisai

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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Currently available inducible Cre/loxP systems, despite their considerable utility in gene manipulation, have pitfalls in certain scenarios, such as unsatisfactory recombination rates and deleterious effects on physiology and behavior. To overcome these limitations, we designed a new, inducible gene-targeting system by introducing an in-frame nonsense mutation into the coding sequence of Cre recombinase (nsCre). Mutant mRNAs transcribed from nsCre transgene can be efficiently translated into full-length, functional Cre recombinase in the presence of nonsense suppressors such as aminoglycosides. In a proof-of-concept model, GABA signaling from hypothalamic neurons expressing agouti-related peptide (AgRP) was genetically inactivated within 4 d after treatment with a synthetic aminoglycoside. Disruption of GABA synthesis in AgRP neurons in young adult mice led to a dramatic loss of body weight due to reduced food intake and elevated energy expenditure; they also manifested glucose intolerance. In contrast, older mice with genetic inactivation of GABA signaling by AgRP neurons had only transient reduction of feeding and body weight; their energy expenditure and glucose tolerance were unaffected. These results indicate that GABAergic signaling from AgRP neurons plays a key role in the control of feeding and metabolism through an age-dependent mechanism. This new genetic technique will augment current tools used to elucidate mechanisms underlying many physiological and neurological processes.

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Mouse Insulin Cells Expressing an Inducible RIPCre Transgene Are Functionally Impaired

Cited by 10 publications

References 29 publications

Phenotypic Characterization of MIP-CreERT1Lphi Mice With Transgene-Driven Islet Expression of Human Growth Hormone

Phenotypic Characterization of MIP-CreERT1Lphi Mice With Transgene-Driven Islet Expression of Human Growth Hormone

lck-Driven Cre Expression Alters T Cell Development in the Thymus and the Frequencies and Functions of Peripheral T Cell Subsets

New inducible genetic method reveals critical roles of GABA in the control of feeding and metabolism

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