A Method for Mouse Pancreatic Islet Isolation and Intracellular cAMP Determination

Neuman, Joshua C.; Truchan, Nathan A.; Joseph, Jamie W.; Kimple, Michelle E.

doi:10.3791/50374

Cited by 45 publications

(33 citation statements)

References 28 publications

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“…Islets were isolated and an ex vivo glucose stimulated insulin secretion assay was performed as described previously (Neuman et al . ; Truchan et al . ; Fontana et al .…”

Section: Methodsmentioning

confidence: 99%

Restoration of metabolic health by decreased consumption of branched‐chain amino acids

Cummings

Williams

Kasza

et al. 2017

The Journal of Physiology

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254

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Key pointsr We recently found that feeding healthy mice a diet with reduced levels of branched-chain amino acids (BCAAs), which are associated with insulin resistance in both humans and rodents, modestly improves glucose tolerance and slows fat mass gain.r In the present study, we show that a reduced BCAA diet promotes rapid fat mass loss without calorie restriction in obese mice.r Selective reduction of dietary BCAAs also restores glucose tolerance and insulin sensitivity to obese mice, even as they continue to consume a high-fat, high-sugar diet.r A low BCAA diet transiently induces FGF21 (fibroblast growth factor 21) and increases energy expenditure.r We suggest that dietary protein quality (i.e. the precise macronutrient composition of dietary protein) may impact the effectiveness of weight loss diets.Abstract Obesity and diabetes are increasing problems around the world, and although even moderate weight loss can improve metabolic health, reduced calorie diets are notoriously difficult to sustain. Branched-chain amino acids (BCAAs; leucine, isoleucine and valine) are elevated in the blood of obese, insulin-resistant humans and rodents. We recently demonstrated that specifically reducing dietary levels of BCAAs has beneficial effects on the metabolic health of young, growing mice, improving glucose tolerance and modestly slowing fat mass gain. In the present study, we examine the hypothesis that reducing dietary BCAAs will promote weight loss, reduce adiposity, and improve blood glucose control in diet-induced obese mice with pre-existing metabolic syndrome. We find that specifically reducing dietary BCAAs rapidly reverses diet-induced obesity and improves glucoregulatory control in diet-induced obese mice. Most dramatically, mice eating an otherwise unhealthy high-calorie, high-sugar Western diet with reduced levels of BCAAs lost weight and fat mass rapidly until regaining a normal weight. Importantly, this normalization of weight was mediated not by caloric restriction or increased activity, but by increased energy expenditure, and was accompanied by a transient induction of the energy balance regulating hormone FGF21 (fibroblast growth factor 21). Consumption of a Western diet reduced in BCAAs was also accompanied by a dramatic improvement in glucose tolerance and insulin resistance. Our results link dietary BCAAs with the regulation of metabolic health and energy balance in obese animals, and suggest that specifically reducing dietary BCAAs may represent a highly translatable option for the treatment of obesity and insulin resistance.

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“…Islets were isolated and an ex vivo glucose stimulated insulin secretion assay was performed as described previously (Neuman et al . ; Truchan et al . ; Fontana et al .…”

Section: Methodsmentioning

confidence: 99%

Restoration of metabolic health by decreased consumption of branched‐chain amino acids

Cummings

Williams

Kasza

et al. 2017

The Journal of Physiology

Self Cite

254

241

View full text Add to dashboard Cite

show abstract

“…Islets were isolated as previously described (Neuman et al, 2014). Full details of the ex vivo glucose stimulated insulin secretion (GSIS) assay and metabolic imaging are provided in Supplemental Experimental Procedures.…”

Section: Methodsmentioning

confidence: 99%

Decreased Consumption of Branched-Chain Amino Acids Improves Metabolic Health

et al. 2016

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Protein restricted, high carbohydrate diets improve metabolic health in rodents, yet the precise dietary components that are responsible for these effects have not been identified. Further, the applicability of these studies to humans is unclear. Here, we demonstrate in a randomized controlled trial that a moderately protein restricted (PR) diet also improves markers of metabolic health in humans. Intriguingly, we find that feeding mice a diet specifically reduced in branched chain amino acids (BCAAs) is sufficient to improve glucose tolerance and body composition equivalently to a PR diet, via metabolically distinct pathways. Our results highlight a critical role for dietary quality at the level of amino acids in the maintenance of metabolic health, and suggest that diets specifically reduced in BCAAs, or pharmacological interventions in this pathway, may offer a translatable way to achieve many of the metabolic benefits of a PR diet.

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“…Islets were isolated and an ex vivo glucose‐stimulated insulin secretion assay was performed (29, 31). Isolated islets were transferred to a 96‐well V‐bottom plate and incubated with RPMI 1640 medium with 10% fetal bovine serum, penicillin‐streptomycin, and 11.1 mM glucose for 48 h (32).…”

Section: Methodsmentioning

confidence: 99%

“…Islets were isolated and an ex vivo glucose-stimulated insulin secretion assay was performed (29,31). Isolated islets were transferred to a 96-well V-bottom plate and incubated with…”

Section: Islet Isolation and Ex Vivo Studiesmentioning

confidence: 99%

Short‐term methionine deprivation improves metabolic health via sexually dimorphic, mTORCI‐independent mechanisms

Yang

Miller

et al. 2018

FASEB j.

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Obesity and diabetes are major challenges to global health, and there is an urgent need for interventions that promote weight loss. Dietary restriction of methionine promotes leanness and improves metabolic health in mice and humans. However, poor long-term adherence to this diet limits its translational potential. In this study, we develop a short-term methionine deprivation (MD) regimen that preferentially reduces fat mass, restoring normal body weight and glycemic control to diet-induced obese mice of both sexes. The benefits of MD do not accrue from calorie restriction, but instead result from increased energy expenditure. MD promotes increased energy expenditure in a sex-specific manner, inducing the fibroblast growth factor (Fgf)-21-uncoupling protein (Ucp)-1 axis only in males. Methionine is an agonist of the protein kinase mechanistic target of rapamycin complex (mTORC)-1, which has been proposed to play a key role in the metabolic response to amino acid-restricted diets. In our study, we used a mouse model of constitutive hepatic mTORC1 activity and demonstrate that suppression of hepatic mTORC1 signaling is not required for the metabolic effects of MD. Our study sheds new light on the mechanisms by which dietary methionine regulates metabolic health and demonstrates the translational potential of MD for the treatment of obesity and type 2 diabetes.-Yu, D., Yang, S. E., Miller, B. R., Wisinski, J. A., Sherman, D. S., Brinkman, J. A., Tomasiewicz, J. L., Cummings, N. E., Kimple, M. E., Cryns, V. L., Lamming, D. W. Short-term methionine deprivation improves metabolic health via sexually dimorphic, mTORC1-independent mechanisms.

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A Method for Mouse Pancreatic Islet Isolation and Intracellular cAMP Determination

Cited by 45 publications

References 28 publications

Restoration of metabolic health by decreased consumption of branched‐chain amino acids

Restoration of metabolic health by decreased consumption of branched‐chain amino acids

Decreased Consumption of Branched-Chain Amino Acids Improves Metabolic Health

Short‐term methionine deprivation improves metabolic health via sexually dimorphic, mTORCI‐independent mechanisms

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