In pancreatic ␤-cells, glucose metabolism signals insulin secretion by altering the cellular array of messenger molecules. ATP is particularly important, given its role in regulating cation channel activity, exocytosis, and events dependent upon its hydrolysis. Uncoupling protein (UCP)-2 is proposed to catalyze a mitochondrial inner-membrane H ؉ leak that bypasses ATP synthase, thereby reducing cellular ATP content. Previously, we showed that overexpression of UCP-2 suppressed glucose-stimulated insulin secretion (GSIS) in isolated islets (1). The aim of this study was to identify downstream consequences of UCP-2 overexpression and to determine whether insufficient insulin secretion in a diabetic model was correlated with increased endogenous UCP-2 expression. In isolated islets from normal rats, the degree to which GSIS was suppressed was inversely correlated with the amount of UCP-2 expression induced. Depolarizing the islets with KCl or inhibiting ATP-dependent K ؉ (K ATP ) channels with glybenclamide elicited similar insulin secretion in control and UCP-2-overexpressing islets. The glucose-stimulated mitochondrial membrane (⌿ m ) hyperpolarization was reduced in ␤-cells overexpressing UCP-2. ATP content of UCP-2-induced islets was reduced by 50%, and there was no change in the efflux of Rb ؉ at high versus low glucose concentrations, suggesting that low ATP led to reduced glucose-induced depolarization, thereby causing reduced insulin secretion. Sprague-Dawley rats fed a diet with 40% fat for 3 weeks were glucose intolerant, and in vitro insulin secretion at high glucose was only increased 8.5-fold over basal, compared with 28-fold in control rats. Islet UCP-2 mRNA expression was increased twofold. These studies provide further strong evidence that UCP-2 is an important negative regulator of ␤-cell insulin secretion and demonstrate that reduced ⌬⌿ m and increased activity of K ATP channels are mechanisms by which UCP-2-mediated effects are mediated. These studies also raise the possibility that a pathological upregulation of UCP-2 expression in the prediabetic state could contribute to the loss of glucose responsiveness observed in obesity-related type 2 diabetes in humans.

We investigated whether an increase in cAMP could normalize glucose-stimulated insulin secretion (GSIS) in uncoupling protein-2 (UCP2) overexpressing (ucp2-OE) bcells. Indices of b-cell (b-TC-6f7 cells and rodent islets) function were measured after induction of ucp2, in the presence or absence of cAMP-stimulating agents, analogs, or inhibitors. Islets of ob/ob mice had improved glucoseresponsiveness in the presence of forskolin. Rat islets overexpressing ucp2 had significantly lower GSIS than controls. Acutely, the protein kinase A (PKA) and epac pathway stimulant forskolin normalized insulin secretion in ucp2-OE rat islets and b-TC-6f7 b-cells, an effect blocked by specific PKA inhibitors but not mimicked by epac agonists. However, there was no effect of ucp2-OE on cAMP concentrations or PKA activity. In ucp2-OE islets, forskolin inhibited ATP-dependent potassium (K ATP ) channel currents and 86 Rb C efflux, indicative of K ATP block. Likewise, forskolin application increased intracellular Ca 2C , which could account for its stimulatory effects on insulin secretion. Chronic exposure to forskolin increased ucp2 mRNA and exaggerated basal secretion but not GSIS. In mice deficient in UCP2, there was no augmentation of either cAMP content or cAMP-dependent insulin secretion. Thus, elevating cellular cAMP can reverse the deficiency in GSIS invoked by ucp2-OE, at least partly through PKA-mediated effects on the K ATP channel.

Multiwavelength images from the Hubble Space Telescope covering the wavelength range 0.27–1.6 μm show that the central area of the nearby dwarf galaxy NGC 4449 contains several tens of compact sources that are emitting in the hydrogen recombination line Paβ (1.2818 μm) but are only marginally detected in Hα (0.6563 μm) and undetected at wavelengths λ ≤ 0.55 μm. An analysis of the spectral energy distributions (SEDs) of these sources indicates that they are likely relatively young stellar clusters heavily attenuated by dust. The selection function used to identify the sources prevents meaningful statistical analyses of their age, mass, and dust extinction distributions. However, these cluster candidates have ages ∼5–6 Myr and A V > 6 mag, according to their SED fits, and are extremely compact, with typical deconvolved radii of 1 pc. The dusty clusters are located at the periphery of the dark clouds within the galaxy and appear to be partially embedded. Density and pressure considerations indicate that the H ii regions surrounding these clusters may be stalled, and that pre-supernova (pre-SN) feedback has not been able to clear the clusters of their natal cocoons. These findings are in potential tension with existing models that regulate star formation with pre-SN feedback, since pre-SN feedback acts on short timescales, ≲4 Myr, for a standard stellar initial mass function. The existence of a population of dusty stellar clusters with ages >4 Myr, if confirmed by future observations, paints a more complex picture for the role of stellar feedback in controlling star formation.

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