ABSTRACT
Hypoglycemia initiates the counter-regulatory response (CRR), in which the sympathetic nervous system, glucagon and glucocorticoids restore glucose to appropriate concentrations. During starvation, low leptin levels restrain energy utilization, enhancing long-term survival. To ensure short-term survival during hypoglycemia in fasted animals, the CRR must overcome this energy-sparing program and nutrient depletion. Here we identify in mice a previously unrecognized role for leptin and a population of leptin-regulated neurons that modulate the CRR to meet these challenges. Hypoglycemia activates neurons of the parabrachial nucleus (PBN) that coexpress leptin receptor (LepRb) and cholecystokinin (CCK) (PBN LepRb(CCK) neurons), which project to the ventromedial hypothalamic nucleus. Leptin inhibits these cells, and Cck(cre)-mediated ablation of LepRb enhances the CRR. Inhibition of PBN LepRb cells blunts the CRR, whereas their activation mimics the CRR in a CCK-dependent manner. PBN LepRb(CCK) neurons are a crucial component of the CRR system and may be a therapeutic target in hypoglycemia.
Subject(s)
Blood Glucose/metabolism , Energy Metabolism/physiology , Hypoglycemia/metabolism , Leptin/pharmacology , Neurons/metabolism , Parabrachial Nucleus/metabolism , Animals , Blood Glucose/drug effects , Energy Metabolism/drug effects , Female , Leptin/physiology , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Neurons/drug effects , Organ Culture Techniques , Parabrachial Nucleus/drug effects , Receptors, Leptin/metabolismABSTRACT
2,4,6-Tribromophenol (2,4,6-TBP) has been used as a wood preservative and flame retardant and is a synthetic intermediate of the most important brominated flame retardants (BFR) produced. The use of TBP-contaminated wood materials in the food industry poses a risk of significant economical losses due to food contamination. In this work an efficient and reliable immunochemical method for analysis of TBP in wood samples has been established consisting of alkaline wood extraction followed by analysis on a microplate ELISA (enzyme-linked immunosorbent assay). TBP is efficiently extracted from wood samples in 10 min and directly measured after 10-fold buffer dilution to avoid matrix interferences. The analytical procedure has a limit of detection of 45 ng g (-1) of TBP in wood (1.5 microg L (-1) in extracts). The method has been applied to the analysis of contaminated real wood samples, showing that the levels of contamination can reach high TBP concentrations (up to 2000 microg L (-1)). An excellent correlation was observed between TBP levels in wood extracts determined by ELISA and gas chromatography-mass spectrometry (GC-MS) analysis ( R (2) = 0.990, N = 19). The precision found is below 22% CV. The immunoanalytical method developed is fast, reliable, and cost-effective, shows good high-throughput screening capabilities, and can be an excellent tool for assessment of wood contamination at lumber mills or related industries. The TBP ELISA has the potential to be used for environmental, food, and biological monitoring of brominated phenolic compounds considered nowadays as emerging pollutants.
