Differential expression of glutamic acid decarboxylase in rat and human islets
Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
Standard
Differential expression of glutamic acid decarboxylase in rat and human islets. / Petersen, J S; Russel, S; Marshall, M O; Kofod, Hans; Buschard, K; Cambon, N; Karlsen, Alan E; Boel, E; Hagopian, W A; Hejnaes, K R.
I: Diabetes, Bind 42, Nr. 3, 03.1993, s. 484-95.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
Harvard
APA
Vancouver
Author
Bibtex
}
RIS
TY - JOUR
T1 - Differential expression of glutamic acid decarboxylase in rat and human islets
AU - Petersen, J S
AU - Russel, S
AU - Marshall, M O
AU - Kofod, Hans
AU - Buschard, K
AU - Cambon, N
AU - Karlsen, Alan E
AU - Boel, E
AU - Hagopian, W A
AU - Hejnaes, K R
PY - 1993/3
Y1 - 1993/3
N2 - The GABA synthesizing enzyme GAD is a prominent islet cell autoantigen in type I diabetes. The two forms of GAD (GAD64 and GAD67) are encoded by different genes in both rats and humans. By in situ hybridization analysis of rat and human pancreases, expression of both genes was detected in rat islets, whereas only GAD64 mRNA was detected in human islets. Immunocytochemical analysis of rat and human pancreatic sections or isolated islets with antibodies to GAD64 and GAD67 in combination with antibodies to insulin, glucagon, or SRIF confirmed that a GAD64 and GAD67 expression were beta-cell specific in rat islets. In contrast, only GAD64 was detected in human islets and was, in addition to beta-cells, also surprisingly localized to some alpha-cells, delta-cells, and PP-cells. In long-term (4 wk) monolayer cultures of newborn rat islet cells, GAD64 expression remained beta-cell specific as observed in vivo, whereas GAD67 was localized not only to the beta-cells but also in the alpha-cells and delta-cells. A small but distinct fraction of GAD positive cells in these monolayer cultures did not accumulate GABA immunoreactivity, which may indicate cellular heterogeneity with respect to GABA catabolism or GAD enzyme activity. In a rat insulinoma cell line (NHI-6F) producing both glucagon and insulin depending on the culture conditions, GAD64 expression was detected only in cultures in which the insulin producing phenotype dominated. In conclusion, these data demonstrate that the two GAD isoforms are differentially expressed in rat and human islets but also that the expression differs according to culture conditions. These findings emphasize the need to consider both the species and culture conditions of islets.
AB - The GABA synthesizing enzyme GAD is a prominent islet cell autoantigen in type I diabetes. The two forms of GAD (GAD64 and GAD67) are encoded by different genes in both rats and humans. By in situ hybridization analysis of rat and human pancreases, expression of both genes was detected in rat islets, whereas only GAD64 mRNA was detected in human islets. Immunocytochemical analysis of rat and human pancreatic sections or isolated islets with antibodies to GAD64 and GAD67 in combination with antibodies to insulin, glucagon, or SRIF confirmed that a GAD64 and GAD67 expression were beta-cell specific in rat islets. In contrast, only GAD64 was detected in human islets and was, in addition to beta-cells, also surprisingly localized to some alpha-cells, delta-cells, and PP-cells. In long-term (4 wk) monolayer cultures of newborn rat islet cells, GAD64 expression remained beta-cell specific as observed in vivo, whereas GAD67 was localized not only to the beta-cells but also in the alpha-cells and delta-cells. A small but distinct fraction of GAD positive cells in these monolayer cultures did not accumulate GABA immunoreactivity, which may indicate cellular heterogeneity with respect to GABA catabolism or GAD enzyme activity. In a rat insulinoma cell line (NHI-6F) producing both glucagon and insulin depending on the culture conditions, GAD64 expression was detected only in cultures in which the insulin producing phenotype dominated. In conclusion, these data demonstrate that the two GAD isoforms are differentially expressed in rat and human islets but also that the expression differs according to culture conditions. These findings emphasize the need to consider both the species and culture conditions of islets.
KW - Animals
KW - Antibodies, Monoclonal
KW - Autoantigens
KW - Base Sequence
KW - Blotting, Western
KW - Fluorescent Antibody Technique
KW - Gene Expression
KW - Glutamate Decarboxylase
KW - Humans
KW - Immunohistochemistry
KW - In Situ Hybridization
KW - Islets of Langerhans
KW - Molecular Sequence Data
KW - Rats
KW - Rats, Inbred Lew
KW - Species Specificity
M3 - Journal article
C2 - 8432419
VL - 42
SP - 484
EP - 495
JO - Diabetes
JF - Diabetes
SN - 0012-1797
IS - 3
ER -
ID: 45574526