Why is it so difficult to measure glucagon-like peptide-1 in a mouse?

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Why is it so difficult to measure glucagon-like peptide-1 in a mouse? / Windeløv, Johanne A; Wewer Albrechtsen, Nicolai J; Kuhre, Rune E; Jepsen, Sara L; Hornburg, Daniel; Pedersen, Jens; Jensen, Elisa P; Galsgaard, Katrine D; Winther-Sørensen, Marie; Ørgaard, Anne; Deacon, Carolyn F; Mann, Matthias; Kissow, Hannelouise; Hartmann, Bolette; Holst, Jens J.

In: Diabetologia, Vol. 60, No. 10, 10.2017, p. 2066-2075.

Research output: Contribution to journalJournal articleResearchpeer-review

Harvard

Windeløv, JA, Wewer Albrechtsen, NJ, Kuhre, RE, Jepsen, SL, Hornburg, D, Pedersen, J, Jensen, EP, Galsgaard, KD, Winther-Sørensen, M, Ørgaard, A, Deacon, CF, Mann, M, Kissow, H, Hartmann, B & Holst, JJ 2017, 'Why is it so difficult to measure glucagon-like peptide-1 in a mouse?', Diabetologia, vol. 60, no. 10, pp. 2066-2075. https://doi.org/10.1007/s00125-017-4347-7

APA

Windeløv, J. A., Wewer Albrechtsen, N. J., Kuhre, R. E., Jepsen, S. L., Hornburg, D., Pedersen, J., ... Holst, J. J. (2017). Why is it so difficult to measure glucagon-like peptide-1 in a mouse? Diabetologia, 60(10), 2066-2075. https://doi.org/10.1007/s00125-017-4347-7

Vancouver

Windeløv JA, Wewer Albrechtsen NJ, Kuhre RE, Jepsen SL, Hornburg D, Pedersen J et al. Why is it so difficult to measure glucagon-like peptide-1 in a mouse? Diabetologia. 2017 Oct;60(10):2066-2075. https://doi.org/10.1007/s00125-017-4347-7

Author

Windeløv, Johanne A ; Wewer Albrechtsen, Nicolai J ; Kuhre, Rune E ; Jepsen, Sara L ; Hornburg, Daniel ; Pedersen, Jens ; Jensen, Elisa P ; Galsgaard, Katrine D ; Winther-Sørensen, Marie ; Ørgaard, Anne ; Deacon, Carolyn F ; Mann, Matthias ; Kissow, Hannelouise ; Hartmann, Bolette ; Holst, Jens J. / Why is it so difficult to measure glucagon-like peptide-1 in a mouse?. In: Diabetologia. 2017 ; Vol. 60, No. 10. pp. 2066-2075.

Bibtex

@article{1c2dfeffef51404eba2e21bc03527a9f,
title = "Why is it so difficult to measure glucagon-like peptide-1 in a mouse?",
abstract = "AIMS/HYPOTHESIS: In humans, glucagon-like peptide-1 (GLP-1) is rapidly degraded by dipeptidyl peptidase-4 to a relatively stable metabolite, GLP-1(9-36)NH2, which allows measurement of GLP-1 secretion. However, little is known about the kinetics of the GLP-1 metabolite in mice. We hypothesised that the GLP-1 metabolite is rapidly degraded in this species by neutral endopeptidase(s) (NEP[s]).METHODS: We administered glucose, mixed meal or water orally to 256 mice, and took blood samples before and 2, 6, 10, 20, 30, 60 or 90 min after stimulation. To study the metabolism of the GLP-1 metabolite, i.v. GLP-1(9-36)NH2 (800 fmol) or saline (154 mmol/l NaCl) was administered to 160 mice, some of which had a prior injection of a selective NEP 24.11 ± inhibitor (candoxatril, 5 mg/kg) or saline. Blood was collected before and 1, 2, 4 and 12 min after GLP-1/saline injection. Plasma GLP-1 levels were analysed using a customised single-site C-terminal ELISA, two different two-site ELISAs and MS.RESULTS: GLP-1 secretion profiles after oral glucose administration differed markedly when assayed by C-terminal ELISA compared with sandwich ELISAs, with the former showing a far higher peak value and AUC. In mice injected with GLP-1(9-36)NH2, immunoreactive GLP-1 plasma levels peaked at approximately 75 pmol/l at 1 min when measured with sandwich ELISAs, returning to baseline (~20 pmol/l) after 12 min, but remained elevated using the C-terminal ELISA (~90 pmol/l at 12 min). NEP 24.11 inhibition by candoxatril significantly attenuated GLP-1(9-36)NH2 degradation in vivo and in vitro. MS identified GLP-1 fragments consistent with NEP 24.11 degradation.CONCLUSIONS/INTERPRETATION: In mice, the GLP-1 metabolite is eliminated within a few minutes owing to endoproteolytic cleavage by NEP 24.11. Therefore, accurate measurement of GLP-1 secretion in mice requires assays for NEP 24.11 metabolites. Conventional sandwich ELISAs are inadequate because of endoproteolytic cleavage of the dipeptidyl peptidase-4-generated metabolite.",
keywords = "Journal Article",
author = "Windel{\o}v, {Johanne A} and {Wewer Albrechtsen}, {Nicolai J} and Kuhre, {Rune E} and Jepsen, {Sara L} and Daniel Hornburg and Jens Pedersen and Jensen, {Elisa P} and Galsgaard, {Katrine D} and Marie Winther-S{\o}rensen and Anne {\O}rgaard and Deacon, {Carolyn F} and Matthias Mann and Hannelouise Kissow and Bolette Hartmann and Holst, {Jens J}",
year = "2017",
month = "10",
doi = "10.1007/s00125-017-4347-7",
language = "English",
volume = "60",
pages = "2066--2075",
journal = "Diabetologia",
issn = "0012-186X",
publisher = "Springer",
number = "10",

}

RIS

TY - JOUR

T1 - Why is it so difficult to measure glucagon-like peptide-1 in a mouse?

AU - Windeløv, Johanne A

AU - Wewer Albrechtsen, Nicolai J

AU - Kuhre, Rune E

AU - Jepsen, Sara L

AU - Hornburg, Daniel

AU - Pedersen, Jens

AU - Jensen, Elisa P

AU - Galsgaard, Katrine D

AU - Winther-Sørensen, Marie

AU - Ørgaard, Anne

AU - Deacon, Carolyn F

AU - Mann, Matthias

AU - Kissow, Hannelouise

AU - Hartmann, Bolette

AU - Holst, Jens J

PY - 2017/10

Y1 - 2017/10

N2 - AIMS/HYPOTHESIS: In humans, glucagon-like peptide-1 (GLP-1) is rapidly degraded by dipeptidyl peptidase-4 to a relatively stable metabolite, GLP-1(9-36)NH2, which allows measurement of GLP-1 secretion. However, little is known about the kinetics of the GLP-1 metabolite in mice. We hypothesised that the GLP-1 metabolite is rapidly degraded in this species by neutral endopeptidase(s) (NEP[s]).METHODS: We administered glucose, mixed meal or water orally to 256 mice, and took blood samples before and 2, 6, 10, 20, 30, 60 or 90 min after stimulation. To study the metabolism of the GLP-1 metabolite, i.v. GLP-1(9-36)NH2 (800 fmol) or saline (154 mmol/l NaCl) was administered to 160 mice, some of which had a prior injection of a selective NEP 24.11 ± inhibitor (candoxatril, 5 mg/kg) or saline. Blood was collected before and 1, 2, 4 and 12 min after GLP-1/saline injection. Plasma GLP-1 levels were analysed using a customised single-site C-terminal ELISA, two different two-site ELISAs and MS.RESULTS: GLP-1 secretion profiles after oral glucose administration differed markedly when assayed by C-terminal ELISA compared with sandwich ELISAs, with the former showing a far higher peak value and AUC. In mice injected with GLP-1(9-36)NH2, immunoreactive GLP-1 plasma levels peaked at approximately 75 pmol/l at 1 min when measured with sandwich ELISAs, returning to baseline (~20 pmol/l) after 12 min, but remained elevated using the C-terminal ELISA (~90 pmol/l at 12 min). NEP 24.11 inhibition by candoxatril significantly attenuated GLP-1(9-36)NH2 degradation in vivo and in vitro. MS identified GLP-1 fragments consistent with NEP 24.11 degradation.CONCLUSIONS/INTERPRETATION: In mice, the GLP-1 metabolite is eliminated within a few minutes owing to endoproteolytic cleavage by NEP 24.11. Therefore, accurate measurement of GLP-1 secretion in mice requires assays for NEP 24.11 metabolites. Conventional sandwich ELISAs are inadequate because of endoproteolytic cleavage of the dipeptidyl peptidase-4-generated metabolite.

AB - AIMS/HYPOTHESIS: In humans, glucagon-like peptide-1 (GLP-1) is rapidly degraded by dipeptidyl peptidase-4 to a relatively stable metabolite, GLP-1(9-36)NH2, which allows measurement of GLP-1 secretion. However, little is known about the kinetics of the GLP-1 metabolite in mice. We hypothesised that the GLP-1 metabolite is rapidly degraded in this species by neutral endopeptidase(s) (NEP[s]).METHODS: We administered glucose, mixed meal or water orally to 256 mice, and took blood samples before and 2, 6, 10, 20, 30, 60 or 90 min after stimulation. To study the metabolism of the GLP-1 metabolite, i.v. GLP-1(9-36)NH2 (800 fmol) or saline (154 mmol/l NaCl) was administered to 160 mice, some of which had a prior injection of a selective NEP 24.11 ± inhibitor (candoxatril, 5 mg/kg) or saline. Blood was collected before and 1, 2, 4 and 12 min after GLP-1/saline injection. Plasma GLP-1 levels were analysed using a customised single-site C-terminal ELISA, two different two-site ELISAs and MS.RESULTS: GLP-1 secretion profiles after oral glucose administration differed markedly when assayed by C-terminal ELISA compared with sandwich ELISAs, with the former showing a far higher peak value and AUC. In mice injected with GLP-1(9-36)NH2, immunoreactive GLP-1 plasma levels peaked at approximately 75 pmol/l at 1 min when measured with sandwich ELISAs, returning to baseline (~20 pmol/l) after 12 min, but remained elevated using the C-terminal ELISA (~90 pmol/l at 12 min). NEP 24.11 inhibition by candoxatril significantly attenuated GLP-1(9-36)NH2 degradation in vivo and in vitro. MS identified GLP-1 fragments consistent with NEP 24.11 degradation.CONCLUSIONS/INTERPRETATION: In mice, the GLP-1 metabolite is eliminated within a few minutes owing to endoproteolytic cleavage by NEP 24.11. Therefore, accurate measurement of GLP-1 secretion in mice requires assays for NEP 24.11 metabolites. Conventional sandwich ELISAs are inadequate because of endoproteolytic cleavage of the dipeptidyl peptidase-4-generated metabolite.

KW - Journal Article

U2 - 10.1007/s00125-017-4347-7

DO - 10.1007/s00125-017-4347-7

M3 - Journal article

C2 - 28669086

VL - 60

SP - 2066

EP - 2075

JO - Diabetologia

JF - Diabetologia

SN - 0012-186X

IS - 10

ER -

ID: 182932367