Insulin storage and glucose homeostasis in mice null for the granule zinc transporter ZnT8 and studies of the type 2 diabetes-associated variants

Tamara J Nicolson; Elisa A Bellomo; Nadeeja Wijesekara; Merewyn K Loder; Jocelyn M Baldwin; Armen V Gyulkhandanyan; Vasilij Koshkin; Andrei I Tarasov; Raffaella Carzaniga; Katrin Kronenberger; Tarvinder K Taneja; Gabriela da Silva Xavier; Sarah Libert; Philippe Froguel; Raphael Scharfmann; Volodymir Stetsyuk; Philippe Ravassard; Helen Parker; Fiona M Gribble; Frank Reimann; Robert Sladek; Stephen J Hughes; Paul R V Johnson; Myriam Masseboeuf; Remy Burcelin; Stephen A Baldwin; Ming Liu; Roberto Lara-Lemus; Peter Arvan; Frans C Schuit; Michael B Wheeler; Fabrice Chimienti; Guy A Rutter

doi:10.2337/db09-0551

Insulin storage and glucose homeostasis in mice null for the granule zinc transporter ZnT8 and studies of the type 2 diabetes-associated variants

Tamara J Nicolson, Elisa A Bellomo, Nadeeja Wijesekara, Merewyn K Loder, Jocelyn M Baldwin, Armen V Gyulkhandanyan, Vasilij Koshkin, Andrei I Tarasov, Raffaella Carzaniga, Katrin Kronenberger, Tarvinder K Taneja, Gabriela da Silva Xavier, Sarah Libert, Philippe Froguel, Raphael Scharfmann, Volodymir Stetsyuk, Philippe Ravassard, Helen Parker, Fiona M Gribble, Frank ReimannRobert Sladek, Stephen J Hughes, Paul R V Johnson, Myriam Masseboeuf, Remy Burcelin, Stephen A Baldwin, Ming Liu, Roberto Lara-Lemus, Peter Arvan, Frans C Schuit, Michael B Wheeler, Fabrice Chimienti, Guy A Rutter

School of Life and Medical Sciences

Research output: Contribution to journal › Article › peer-review

286 Citations (Scopus)

Abstract

OBJECTIVE: Zinc ions are essential for the formation of hexameric insulin and hormone crystallization. A nonsynonymous single nucleotide polymorphism rs13266634 in the SLC30A8 gene, encoding the secretory granule zinc transporter ZnT8, is associated with type 2 diabetes. We describe the effects of deleting the ZnT8 gene in mice and explore the action of the at-risk allele.

RESEARCH DESIGN AND METHODS: Slc30a8 null mice were generated and backcrossed at least twice onto a C57BL/6J background. Glucose and insulin tolerance were measured by intraperitoneal injection or euglycemic clamp, respectively. Insulin secretion, electrophysiology, imaging, and the generation of adenoviruses encoding the low- (W325) or elevated- (R325) risk ZnT8 alleles were undertaken using standard protocols.

RESULTS: ZnT8(-/-) mice displayed age-, sex-, and diet-dependent abnormalities in glucose tolerance, insulin secretion, and body weight. Islets isolated from null mice had reduced granule zinc content and showed age-dependent changes in granule morphology, with markedly fewer dense cores but more rod-like crystals. Glucose-stimulated insulin secretion, granule fusion, and insulin crystal dissolution, assessed by total internal reflection fluorescence microscopy, were unchanged or enhanced in ZnT8(-/-) islets. Insulin processing was normal. Molecular modeling revealed that residue-325 was located at the interface between ZnT8 monomers. Correspondingly, the R325 variant displayed lower apparent Zn(2+) transport activity than W325 ZnT8 by fluorescence-based assay.

CONCLUSIONS: ZnT8 is required for normal insulin crystallization and insulin release in vivo but not, remarkably, in vitro. Defects in the former processes in carriers of the R allele may increase type 2 diabetes risks.

Original language	English
Pages (from-to)	2070-83
Number of pages	14
Journal	Diabetes
Volume	58
Issue number	9
DOIs	https://doi.org/10.2337/db09-0551
Publication status	Published - Sept 2009

Keywords

Animals
Blood Glucose/metabolism
Cation Transport Proteins/genetics
Cytoplasmic Granules/metabolism
Diabetes Mellitus, Type 2/epidemiology
Exocytosis/physiology
Female
Gene Expression/physiology
HeLa Cells
Homeostasis/physiology
Humans
Insulin/metabolism
Insulin-Secreting Cells/pathology
Male
Mice
Mice, Inbred C57BL
Mice, Knockout
Polymorphism, Genetic
Risk Factors
Zinc/metabolism
Zinc Transporter 8

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10.2337/db09-0551

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Nicolson, T. J., Bellomo, E. A., Wijesekara, N., Loder, M. K., Baldwin, J. M., Gyulkhandanyan, A. V., Koshkin, V., Tarasov, A. I., Carzaniga, R., Kronenberger, K., Taneja, T. K., da Silva Xavier, G., Libert, S., Froguel, P., Scharfmann, R., Stetsyuk, V., Ravassard, P., Parker, H., Gribble, F. M., ... Rutter, G. A. (2009). Insulin storage and glucose homeostasis in mice null for the granule zinc transporter ZnT8 and studies of the type 2 diabetes-associated variants. Diabetes, 58(9), 2070-83. https://doi.org/10.2337/db09-0551

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title = "Insulin storage and glucose homeostasis in mice null for the granule zinc transporter ZnT8 and studies of the type 2 diabetes-associated variants",

abstract = "OBJECTIVE: Zinc ions are essential for the formation of hexameric insulin and hormone crystallization. A nonsynonymous single nucleotide polymorphism rs13266634 in the SLC30A8 gene, encoding the secretory granule zinc transporter ZnT8, is associated with type 2 diabetes. We describe the effects of deleting the ZnT8 gene in mice and explore the action of the at-risk allele.RESEARCH DESIGN AND METHODS: Slc30a8 null mice were generated and backcrossed at least twice onto a C57BL/6J background. Glucose and insulin tolerance were measured by intraperitoneal injection or euglycemic clamp, respectively. Insulin secretion, electrophysiology, imaging, and the generation of adenoviruses encoding the low- (W325) or elevated- (R325) risk ZnT8 alleles were undertaken using standard protocols.RESULTS: ZnT8(-/-) mice displayed age-, sex-, and diet-dependent abnormalities in glucose tolerance, insulin secretion, and body weight. Islets isolated from null mice had reduced granule zinc content and showed age-dependent changes in granule morphology, with markedly fewer dense cores but more rod-like crystals. Glucose-stimulated insulin secretion, granule fusion, and insulin crystal dissolution, assessed by total internal reflection fluorescence microscopy, were unchanged or enhanced in ZnT8(-/-) islets. Insulin processing was normal. Molecular modeling revealed that residue-325 was located at the interface between ZnT8 monomers. Correspondingly, the R325 variant displayed lower apparent Zn(2+) transport activity than W325 ZnT8 by fluorescence-based assay.CONCLUSIONS: ZnT8 is required for normal insulin crystallization and insulin release in vivo but not, remarkably, in vitro. Defects in the former processes in carriers of the R allele may increase type 2 diabetes risks.",

keywords = "Animals, Blood Glucose/metabolism, Cation Transport Proteins/genetics, Cytoplasmic Granules/metabolism, Diabetes Mellitus, Type 2/epidemiology, Exocytosis/physiology, Female, Gene Expression/physiology, HeLa Cells, Homeostasis/physiology, Humans, Insulin/metabolism, Insulin-Secreting Cells/pathology, Male, Mice, Mice, Inbred C57BL, Mice, Knockout, Polymorphism, Genetic, Risk Factors, Zinc/metabolism, Zinc Transporter 8",

author = "Nicolson, {Tamara J} and Bellomo, {Elisa A} and Nadeeja Wijesekara and Loder, {Merewyn K} and Baldwin, {Jocelyn M} and Gyulkhandanyan, {Armen V} and Vasilij Koshkin and Tarasov, {Andrei I} and Raffaella Carzaniga and Katrin Kronenberger and Taneja, {Tarvinder K} and {da Silva Xavier}, Gabriela and Sarah Libert and Philippe Froguel and Raphael Scharfmann and Volodymir Stetsyuk and Philippe Ravassard and Helen Parker and Gribble, {Fiona M} and Frank Reimann and Robert Sladek and Hughes, {Stephen J} and Johnson, {Paul R V} and Myriam Masseboeuf and Remy Burcelin and Baldwin, {Stephen A} and Ming Liu and Roberto Lara-Lemus and Peter Arvan and Schuit, {Frans C} and Wheeler, {Michael B} and Fabrice Chimienti and Rutter, {Guy A}",

year = "2009",

month = sep,

doi = "10.2337/db09-0551",

language = "English",

volume = "58",

pages = "2070--83",

journal = "Diabetes",

issn = "0012-1797",

publisher = "American Diabetes Association Inc.",

number = "9",

}

Nicolson, TJ, Bellomo, EA, Wijesekara, N, Loder, MK, Baldwin, JM, Gyulkhandanyan, AV, Koshkin, V, Tarasov, AI, Carzaniga, R, Kronenberger, K, Taneja, TK, da Silva Xavier, G, Libert, S, Froguel, P, Scharfmann, R, Stetsyuk, V, Ravassard, P, Parker, H, Gribble, FM, Reimann, F, Sladek, R, Hughes, SJ, Johnson, PRV, Masseboeuf, M, Burcelin, R, Baldwin, SA, Liu, M, Lara-Lemus, R, Arvan, P, Schuit, FC, Wheeler, MB, Chimienti, F & Rutter, GA 2009, 'Insulin storage and glucose homeostasis in mice null for the granule zinc transporter ZnT8 and studies of the type 2 diabetes-associated variants', Diabetes, vol. 58, no. 9, pp. 2070-83. https://doi.org/10.2337/db09-0551

TY - JOUR

T1 - Insulin storage and glucose homeostasis in mice null for the granule zinc transporter ZnT8 and studies of the type 2 diabetes-associated variants

AU - Nicolson, Tamara J

AU - Bellomo, Elisa A

AU - Wijesekara, Nadeeja

AU - Loder, Merewyn K

AU - Baldwin, Jocelyn M

AU - Gyulkhandanyan, Armen V

AU - Koshkin, Vasilij

AU - Tarasov, Andrei I

AU - Carzaniga, Raffaella

AU - Kronenberger, Katrin

AU - Taneja, Tarvinder K

AU - da Silva Xavier, Gabriela

AU - Libert, Sarah

AU - Froguel, Philippe

AU - Scharfmann, Raphael

AU - Stetsyuk, Volodymir

AU - Ravassard, Philippe

AU - Parker, Helen

AU - Gribble, Fiona M

AU - Reimann, Frank

AU - Sladek, Robert

AU - Hughes, Stephen J

AU - Johnson, Paul R V

AU - Masseboeuf, Myriam

AU - Burcelin, Remy

AU - Baldwin, Stephen A

AU - Liu, Ming

AU - Lara-Lemus, Roberto

AU - Arvan, Peter

AU - Schuit, Frans C

AU - Wheeler, Michael B

AU - Chimienti, Fabrice

AU - Rutter, Guy A

PY - 2009/9

Y1 - 2009/9

N2 - OBJECTIVE: Zinc ions are essential for the formation of hexameric insulin and hormone crystallization. A nonsynonymous single nucleotide polymorphism rs13266634 in the SLC30A8 gene, encoding the secretory granule zinc transporter ZnT8, is associated with type 2 diabetes. We describe the effects of deleting the ZnT8 gene in mice and explore the action of the at-risk allele.RESEARCH DESIGN AND METHODS: Slc30a8 null mice were generated and backcrossed at least twice onto a C57BL/6J background. Glucose and insulin tolerance were measured by intraperitoneal injection or euglycemic clamp, respectively. Insulin secretion, electrophysiology, imaging, and the generation of adenoviruses encoding the low- (W325) or elevated- (R325) risk ZnT8 alleles were undertaken using standard protocols.RESULTS: ZnT8(-/-) mice displayed age-, sex-, and diet-dependent abnormalities in glucose tolerance, insulin secretion, and body weight. Islets isolated from null mice had reduced granule zinc content and showed age-dependent changes in granule morphology, with markedly fewer dense cores but more rod-like crystals. Glucose-stimulated insulin secretion, granule fusion, and insulin crystal dissolution, assessed by total internal reflection fluorescence microscopy, were unchanged or enhanced in ZnT8(-/-) islets. Insulin processing was normal. Molecular modeling revealed that residue-325 was located at the interface between ZnT8 monomers. Correspondingly, the R325 variant displayed lower apparent Zn(2+) transport activity than W325 ZnT8 by fluorescence-based assay.CONCLUSIONS: ZnT8 is required for normal insulin crystallization and insulin release in vivo but not, remarkably, in vitro. Defects in the former processes in carriers of the R allele may increase type 2 diabetes risks.

AB - OBJECTIVE: Zinc ions are essential for the formation of hexameric insulin and hormone crystallization. A nonsynonymous single nucleotide polymorphism rs13266634 in the SLC30A8 gene, encoding the secretory granule zinc transporter ZnT8, is associated with type 2 diabetes. We describe the effects of deleting the ZnT8 gene in mice and explore the action of the at-risk allele.RESEARCH DESIGN AND METHODS: Slc30a8 null mice were generated and backcrossed at least twice onto a C57BL/6J background. Glucose and insulin tolerance were measured by intraperitoneal injection or euglycemic clamp, respectively. Insulin secretion, electrophysiology, imaging, and the generation of adenoviruses encoding the low- (W325) or elevated- (R325) risk ZnT8 alleles were undertaken using standard protocols.RESULTS: ZnT8(-/-) mice displayed age-, sex-, and diet-dependent abnormalities in glucose tolerance, insulin secretion, and body weight. Islets isolated from null mice had reduced granule zinc content and showed age-dependent changes in granule morphology, with markedly fewer dense cores but more rod-like crystals. Glucose-stimulated insulin secretion, granule fusion, and insulin crystal dissolution, assessed by total internal reflection fluorescence microscopy, were unchanged or enhanced in ZnT8(-/-) islets. Insulin processing was normal. Molecular modeling revealed that residue-325 was located at the interface between ZnT8 monomers. Correspondingly, the R325 variant displayed lower apparent Zn(2+) transport activity than W325 ZnT8 by fluorescence-based assay.CONCLUSIONS: ZnT8 is required for normal insulin crystallization and insulin release in vivo but not, remarkably, in vitro. Defects in the former processes in carriers of the R allele may increase type 2 diabetes risks.

KW - Animals

KW - Blood Glucose/metabolism

KW - Cation Transport Proteins/genetics

KW - Cytoplasmic Granules/metabolism

KW - Diabetes Mellitus, Type 2/epidemiology

KW - Exocytosis/physiology

KW - Female

KW - Gene Expression/physiology

KW - HeLa Cells

KW - Homeostasis/physiology

KW - Humans

KW - Insulin/metabolism

KW - Insulin-Secreting Cells/pathology

KW - Male

KW - Mice

KW - Mice, Inbred C57BL

KW - Mice, Knockout

KW - Polymorphism, Genetic

KW - Risk Factors

KW - Zinc/metabolism

KW - Zinc Transporter 8

U2 - 10.2337/db09-0551

DO - 10.2337/db09-0551

M3 - Article

C2 - 19542200

SN - 0012-1797

VL - 58

SP - 2070

EP - 2083

JO - Diabetes

JF - Diabetes

IS - 9

ER -

Insulin storage and glucose homeostasis in mice null for the granule zinc transporter ZnT8 and studies of the type 2 diabetes-associated variants

Abstract

Keywords

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