Exposure of pancreatic islets to different alkylating agents decreases mitochondrial DNA content but only streptozotocin induces long-lasting functional impairment of B-cells.
Pancreatic B-cells exposed in vivo or in vitro to streptozotocin (SZ), the N-nitrosourea derivative of glucosamide, present a long-lasting impairment in the production and release of insulin while other cell functions are better preserved. This functional impairment is associated with a defective mitochondrial function. To further study the mechanisms behind SZ actions, mouse pancreatic islets were exposed in vitro to SZ (1.5 mM) or to different concentrations of methyl methanesulfonate (MMS; 2, 4 and 6 mM). The effect of the aglucone moiety of SZ, nitroso-N-methylurea (NMU; 2, 4 and 6 mM) was also tested. Islets were either studied immediately after exposure to the drugs (day 0) or after six days in culture following toxin treatment (day 6). On day 0 the islets showed a decrease in the NAD + NADH content, decreased glucose oxidation rates and an impaired insulin release in response to glucose. Six days after exposure to SZ there was still impaired glucose oxidation and insulin release, and decreased islet insulin mRNA and insulin content, but the NAD + NADH content was again similar to the control group. On the other hand, islets which survived for 6 days in culture following exposure to either MMS or NMU were able to regain normal B-cell function. The mouse islets exposed to SZ, NMU and MMS showed on day 6 a 30-40% decrease in the content of the mitochondrial DNA encoded cytochrome b mRNA and a 60-70% decrease in total mitochondrial DNA, as evaluated by dot and Southern blot analysis. Only SZ decreased the insulin mRNA content whereas both MMS and NMU decreased the glucagon mRNA content. As a whole, the data obtained indicate that SZ, NMU and MMS induce damage to the mitochondrial genome, and this may contribute to the B-cell dysfunction observed after SZ treatment. It is conceivable that the glucose moiety of SZ may direct the methylation to other intracellular sites besides the mitochondrial DNA, thus explaining the different functional responses of islets following exposure to SZ and NMU.
Eizirik
D L
DL
Department of Medical Cell Biology, Uppsala University, Sweden.
Sandler
S
S
Ahnström
G
G
Welsh
M
M
D000225:Adenine / Q000031:analogs & derivatives / Q000032:analysis; D000477:Alkylating Agents / Q000494:pharmacology; D000818:Animals; D003573:Cytochrome b Group / Q000235:genetics; D004247:DNA / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D005934:Glucagon / Q000235:genetics; D005947:Glucose / Q000494:pharmacology; D006147:Guanine / Q000031:analogs & derivatives / Q000032:analysis; D015151:Immunoblotting; D007328:Insulin / Q000032:analysis* / Q000235:genetics; D007515:Islets of Langerhans / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology; D008297:Male; D008741:Methyl Methanesulfonate / Q000494:pharmacology; D008770:Methylnitrosourea / Q000494:pharmacology; D051379:Mice; D008928:Mitochondria / Q000187:drug effects; D012333:RNA, Messenger / Q000032:analysis; D013311:Streptozocin / Q000494:pharmacology*
D000477:Alkylating Agents; D003573:Cytochrome b Group; D007328:Insulin; D012333:RNA, Messenger; C025946:3-methyladenine; D013311:Streptozocin; D006147:Guanine; C008450:7-methylguanine; D008770:Methylnitrosourea; D004247:DNA; D005934:Glucagon; C008449:O-(6)-methylguanine; D008741:Methyl Methanesulfonate; D005947:Glucose; D000225:Adenine