Effects of diabetes and recurrent hypoglycemia on the regulation of the sympathoadrenal system and hypothalamo-pituitary- adrenal (HPA) axis Short Running Title: Sympathoadrenal and HPA regulation in diabetes

Epinephrine, norepinephrine, and corticosterone responses to hypoglycemia are impaired in diabetic rats. Recurrent hypoglycemia further diminishes epinephrine responses. This study examined the sympathoadrenal system and hypothalamo-pituitary-adrenal axis for molecular adaptations underlying these defects. Groups: Normal (N); Diabetic (D); Diabetic exposed to 4 days of 2 episodes/day of hyperinsulinemic-hypoglycemia (D-hypo) or hyperinsulinemichyperglycemia (D-hyper). D-hypo and D-hyper rats differentiated the effects of hypoglycemia and hyperinsulinemia. Adrenal tyrosine hydroxylase (TH) mRNA was reduced (P<0.05 vs. N) by 25% in all diabetic groups. Remarkably, mRNA for phenylethanolamine N-methyltransferase (PNMT), which converts norepinephrine to epinephrine, was reduced (P<0.05 vs. all) by 40% only in D-hypo rats. Paradoxically, dopamine β-hydroxylase mRNA was elevated (P<0.05 vs. D, D-hyper) in D-hypo rats. Hippocampal mineralocorticoid receptor (MR) mRNA was increased (P<0.05 vs. N) in all diabetic groups. Hippocampal glucocorticoid receptor (GR), hypothalamic paraventricular nucleus (PVN) GR and corticotrophin-releasing hormone (CRH), and pituitary GR and pro-opiomelanocortin (POMC) mRNA levels did not differ. We conclude that the blunted corticosterone responses to hypoglycemia in diabetic rats are not due to altered basal expression of GR, CRH, and POMC in the hippocampus, PVN, and pituitary. The corticosterone defect also does not appear to be due to increased hippocampal MR, since we have reported normalized corticosterone responses in D-hypo and D-hyper rats. Further, impaired epinephrine counterregulation in diabetes is associated with reduced adrenal TH mRNA, whereas the additional epinephrine defect after recurrent hypoglycemia is associated with decreases in both TH and PNMT mRNA.