title: β-Cell subcellular localization of glucose-stimulated Mn uptake by X-ray fluorescence microscopy: implications for pancreatic MRI.
abstract: Manganese (Mn) is a calcium (Ca) analog that has long been used as a magnetic resonance imaging (MRI) contrast agent for investigating cardiac tissue functionality, for brain mapping and for neuronal tract tracing studies. Recently, we have extended its use to investigate pancreatic β-cells and showed that, in the presence of MnCl(2), glucose-activated pancreatic islets yield significant signal enhancement in T(1)-weigheted MR images. In this study, we exploited for the first time the unique capabilities of X-ray fluorescence microscopy (XFM) to both visualize and quantify the metal in pancreatic β-cells at cellular and subcellular levels. MIN-6 insulinoma cells grown in standard tissue culture conditions had only a trace amount of Mn, 1.14 ± 0.03 × 10(-11)µg/µm(2), homogenously distributed across the cell. Exposure to 2 mM glucose and 50 µM MnCl(2) for 20 min resulted in nonglucose-dependent Mn uptake and the overall cell concentration increased to 8.99 ± 2.69 × 10(-11) µg/µm(2). When cells were activated by incubation in 16 mM glucose in the presence of 50 µM MnCl(2), a significant increase in cytoplasmic Mn was measured, reaching 2.57 ± 1.34 × 10(-10) µg/µm(2). A further rise in intracellular concentration was measured following KCl-induced depolarization, with concentrations totaling 1.25 ± 0.33 × 10(-9) and 4.02 ± 0.71 × 10(-10) µg/µm(2) in the cytoplasm and nuclei, respectively. In both activated conditions Mn was prevalent in the cytoplasm and localized primarily in a perinuclear region, possibly corresponding to the Golgi apparatus and involving the secretory pathway. These data are consistent with our previous MRI findings, confirming that Mn can be used as a functional imaging reporter of pancreatic β-cell activation and also provide a basis for understanding how subcellular localization of Mn will impact MRI contrast.
authors.affiliation: Department of Radiology, Committee on Medical Physics, University of Chicago, Chicago, IL 60637, USA.
authors.forename: Lara
authors.initials: L
authors.lastname: Leoni
authors.forename: Anita
authors.initials: A
authors.lastname: Dhyani
authors.forename: Patrick
authors.initials: P
authors.lastname: La Riviere
authors.forename: Stefan
authors.initials: S
authors.lastname: Vogt
authors.forename: Barry
authors.initials: B
authors.lastname: Lai
authors.forename: B B
authors.initials: BB
authors.lastname: Roman
mesh_terms: D045744:Cell Line, Tumor; D002712:Chlorides / Q000493:pharmacokinetics*; D003287:Contrast Media / Q000493:pharmacokinetics*; D005947:Glucose / Q000494:pharmacology*; D006801:Humans; D050417:Insulin-Secreting Cells / Q000187:drug effects / Q000378:metabolism*; D008279:Magnetic Resonance Imaging / Q000379:methods*; D017895:Manganese Compounds / Q000493:pharmacokinetics*; D008856:Microscopy, Fluorescence / Q000379:methods*; D010190:Pancreatic Neoplasms / Q000175:diagnosis; D014965:X-Rays
chemical_list: D002712:Chlorides; D003287:Contrast Media; D017895:Manganese Compounds; D005947:Glucose; C025340:manganese chloride