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Regulation of glutamic acid decarboxylase diabetes autoantigen expression in highly purified isolated islets from Macaca nemestrina

Hagopian, W A ; Karlsen, A E ; Petersen, J S ; Teague, J ; Gervassi, A ; Jiang, J ; Fujimoto, W and Lernmark, A LU orcid (1993) In Endocrinology 132(6). p.81-2674
Abstract

Macaca nemestrina, which may have larger and more numerous pancreatic islets than other species, was used for large scale islet isolation by ductal collagenase perfusion and Ficoll gradient centrifugation. The average yield was 51,000 islet equivalents per pancreas, or 8,750 islets equivalents per g. The average purity was 91%, often exceeding 95%. These are the highest reported size, purity, and yield per g of any nonautomated primate islet series. Perifusion with glucose, arginine, and isobutylmethylxanthine showed appropriate biphasic insulin secretion. Unlike that in the rat, human islet glutamic acid decarboxylase (GAD) isoform expression is restricted. However, glycemic regulation of GAD expression has been shown only in rats. We,... (More)

Macaca nemestrina, which may have larger and more numerous pancreatic islets than other species, was used for large scale islet isolation by ductal collagenase perfusion and Ficoll gradient centrifugation. The average yield was 51,000 islet equivalents per pancreas, or 8,750 islets equivalents per g. The average purity was 91%, often exceeding 95%. These are the highest reported size, purity, and yield per g of any nonautomated primate islet series. Perifusion with glucose, arginine, and isobutylmethylxanthine showed appropriate biphasic insulin secretion. Unlike that in the rat, human islet glutamic acid decarboxylase (GAD) isoform expression is restricted. However, glycemic regulation of GAD expression has been shown only in rats. We, therefore, tested hypotheses that M. nemestrina islets also have restricted GAD expression, that GAD expression in primates is stimulated by glucose, and that this stimulation remains restricted to the 64,000 mol wt (GAD65) isoform. Immunoprecipitation of labeled islet extracts showed that GAD65 expression increased 16.7 +/- 0.6-fold during high glucose in vitro culture. After controlling for observed increases in protein synthesis, specific glucose stimulation was still 4.2 +/- 0.2-fold. Specific antisera revealed no GAD67 expression under basal conditions, and isoform restriction was maintained during stimulation. Increased GAD65 synthesis thus accounts for glucose stimulation of 64K expression. These time- and concentration-dependent effects of glucose suggest that hyperglycemia increases autoantigenicity and may accelerate beta-cell destruction in primates, supporting a role for beta-cell rest in insulin-dependent diabetes mellitus prevention.

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author
; ; ; ; ; ; and
publishing date
type
Contribution to journal
publication status
published
keywords
Animals, Autoantigens/immunology, Diabetes Mellitus, Experimental/immunology, Female, Glucose/pharmacology, Glutamate Decarboxylase/immunology, In Vitro Techniques, Insulin/metabolism, Insulin Secretion, Islets of Langerhans/immunology, Isoenzymes/metabolism, Macaca nemestrina, Male, Perfusion/methods
in
Endocrinology
volume
132
issue
6
pages
81 - 2674
publisher
Oxford University Press
external identifiers
  • pmid:8504767
  • scopus:0027215096
ISSN
0013-7227
DOI
10.1210/endo.132.6.8504767
language
English
LU publication?
no
id
08bf12a8-1b73-4179-b498-53723b9f97e5
date added to LUP
2021-09-21 16:08:34
date last changed
2024-03-13 08:13:04
@article{08bf12a8-1b73-4179-b498-53723b9f97e5,
  abstract     = {{<p>Macaca nemestrina, which may have larger and more numerous pancreatic islets than other species, was used for large scale islet isolation by ductal collagenase perfusion and Ficoll gradient centrifugation. The average yield was 51,000 islet equivalents per pancreas, or 8,750 islets equivalents per g. The average purity was 91%, often exceeding 95%. These are the highest reported size, purity, and yield per g of any nonautomated primate islet series. Perifusion with glucose, arginine, and isobutylmethylxanthine showed appropriate biphasic insulin secretion. Unlike that in the rat, human islet glutamic acid decarboxylase (GAD) isoform expression is restricted. However, glycemic regulation of GAD expression has been shown only in rats. We, therefore, tested hypotheses that M. nemestrina islets also have restricted GAD expression, that GAD expression in primates is stimulated by glucose, and that this stimulation remains restricted to the 64,000 mol wt (GAD65) isoform. Immunoprecipitation of labeled islet extracts showed that GAD65 expression increased 16.7 +/- 0.6-fold during high glucose in vitro culture. After controlling for observed increases in protein synthesis, specific glucose stimulation was still 4.2 +/- 0.2-fold. Specific antisera revealed no GAD67 expression under basal conditions, and isoform restriction was maintained during stimulation. Increased GAD65 synthesis thus accounts for glucose stimulation of 64K expression. These time- and concentration-dependent effects of glucose suggest that hyperglycemia increases autoantigenicity and may accelerate beta-cell destruction in primates, supporting a role for beta-cell rest in insulin-dependent diabetes mellitus prevention.</p>}},
  author       = {{Hagopian, W A and Karlsen, A E and Petersen, J S and Teague, J and Gervassi, A and Jiang, J and Fujimoto, W and Lernmark, A}},
  issn         = {{0013-7227}},
  keywords     = {{Animals; Autoantigens/immunology; Diabetes Mellitus, Experimental/immunology; Female; Glucose/pharmacology; Glutamate Decarboxylase/immunology; In Vitro Techniques; Insulin/metabolism; Insulin Secretion; Islets of Langerhans/immunology; Isoenzymes/metabolism; Macaca nemestrina; Male; Perfusion/methods}},
  language     = {{eng}},
  number       = {{6}},
  pages        = {{81--2674}},
  publisher    = {{Oxford University Press}},
  series       = {{Endocrinology}},
  title        = {{Regulation of glutamic acid decarboxylase diabetes autoantigen expression in highly purified isolated islets from Macaca nemestrina}},
  url          = {{http://dx.doi.org/10.1210/endo.132.6.8504767}},
  doi          = {{10.1210/endo.132.6.8504767}},
  volume       = {{132}},
  year         = {{1993}},
}