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Åke Lernmark

Principal investigator

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Differential detection of rat islet and brain glutamic acid decarboxylase (GAD) isoforms with sequence-specific peptide antibodies

Author

  • L. Li
  • J. Jiang
  • W. A. Hagopian
  • A. E. Karlsen
  • M. Skelly
  • D. G. Baskin
  • A. Lernmark

Summary, in English

We studied the distribution of the M(r) 65,000 and M(r) 67,000 isoforms of glutamic acid decarboxylase, GAD65 and GAD67, in rat islets and brain by immunocytochemistry. Synthetic peptides representing selected GAD65 or GAD67 sequences were used to produce sequence-specific antibodies, allowing differential immunocytochemical detection of the two isoforms. GAD-specific reactivity of each peptide antiserum was confirmed by ELISA, immunoblotting, and immunoprecipitation. Immunostaining specificity was verified by displacement with either immunizing or irrelevant peptide. Dual immunostaining with GAD isoform-specific antibodies and polyclonal antibodies to glucagon showed that GAD65 was primarily detected in rat pancreatic islet β-cells, whereas α-cells had weak GAD65 staining. In contrast, GAD67 was detected primarily in α-cells. In rat brain, GAD65 and GAD67 were present in neuron cell bodies and processes. These data demonstrate that antibodies raised against the N-terminus of GAD allow differential immunocytochemical identification of GAD67 and GAD65. Differential expression of GAD isoforms within islet α- and β-cells supports the role of GAD65 in autoimmune diabetes and stiff-man syndrome.

Publishing year

1995-01-01

Language

English

Pages

53-59

Publication/Series

Journal of Histochemistry and Cytochemistry

Volume

43

Issue

1

Document type

Journal article

Publisher

Histochemical Society

Keywords

  • γ-aminobutyric acid, GABA
  • Immunostaining
  • Insulin-dependent diabetes, IDDM
  • Stiff-man syndrome, SMS

Status

Published

ISBN/ISSN/Other

  • ISSN: 0022-1554