Mechanisms of dual modulatory effects of spermine on the mitochondrial calcium uniporter complex.

The mitochondrial Ca2+ uniporter mediates the crucial cellular process of mitochondrial Ca2+ uptake, which regulates cell bioenergetics, intracellular Ca2+ signaling, and cell death initiation. The uniporter contains the pore-forming MCU subunit, an EMRE protein that binds to MCU, and the regulatory MICU1 subunit, which can dimerize with MICU1 or MICU2 and under resting cellular [Ca2+] occludes the MCU pore. It has been known for decades that spermine, which is ubiquitously present in animal cells, can enhance mitochondrial Ca2+ uptake, but the underlying mechanisms remain unclear. Here, we show that spermine exerts dual modulatory effects on the uniporter. In physiological concentrations of spermine, it enhances uniporter activity by breaking the physical interactions between MCU and the MICU1-containing dimers to allow the uniporter to constitutively take up Ca2+ even in low [Ca2+] conditions. This potentiation effect does not require MICU2 or the EF-hand motifs in MICU1. When [spermine] rises to millimolar levels, it inhibits the uniporter by targeting the pore region in a MICU-independent manner. The MICU1-dependent spermine potentiation mechanism proposed here, along with our previous finding that cardiac mitochondria have very low MICU1, can explain the puzzling observation in the literature that mitochondria in the heart show no response to spermine.

Evidence supporting the MICU1 occlusion mechanism and against the potentiation model in the mitochondrial calcium uniporter complex.

The mitochondrial calcium uniporter is a Ca2+ channel that imports cytoplasmic Ca2+ into the mitochondrial matrix to regulate cell bioenergetics, intracellular Ca2+ signaling, and apoptosis. The uniporter contains the pore-forming MCU subunit, an auxiliary EMRE protein, and the regulatory MICU1/MICU2 subunits. Structural and biochemical studies have suggested that MICU1 gates MCU by blocking/unblocking the pore. However, mitoplast patch-clamp experiments argue that MICU1 does not block, but instead potentiates MCU via allosteric mechanisms. Here, we address this direct clash of the proposed MICU1 function. Supporting the MICU1-occlusion mechanism, patch-clamp demonstrates that purified MICU1 strongly suppresses MCU Ca2+ currents, and this inhibition is abolished by mutating the MCU-interacting K126 residue. Moreover, a membrane-depolarization assay shows that MICU1 prevents MCU-mediated Na+ flux into intact mitochondria under Ca2+-free conditions. Examining the observations underlying the potentiation model, we found that MICU1 occlusion was not detected in mitoplasts not because MICU1 cannot block, but because MICU1 dissociates from the uniporter complex. Furthermore, MICU1 depletion reduces uniporter transport not because MICU1 can potentiate MCU, but because EMRE is down-regulated. These results firmly establish the molecular mechanisms underlying the physiologically crucial process of uniporter regulation by MICU1.

Evaluating the Effect of Different Polymer and Composite Abutments on the Color Accuracy of Multilayer Pre-Colored Zirconia Polycrystal Dental Prosthesis.

With increasing aesthetic awareness and emphasis on time costs in today's society, monolithic multilayer precolored zirconia ceramics (M-Zr) facilitate aesthetic restorations in a convenient and straightforward manner without the need for veneering porcelain to modify the color. However, the effect of abutment materials on the final color of M-Zr remains unclear. Herein, we placed Vita A1 Shade M-Zr on six different abutment materials, zirconia (Y-TZP), 3D printed composite resin (CR), dental model resin (MR), polyetheretherketone (PEEK), polyetherketoneketone (PEKK), and cobalt−chromium alloy (Co−Cr), to evaluate their effect on the color accuracy of M-Zr. The color attributes (L*, a*, and b*) were measured using a dental spectrophotometer. The translucency parameter (TP), contrast ratio, color difference (ΔE) between each background substrate and the Vita A1 Shade Guide, and chroma values (C) were calculated to evaluate the color accuracy of M-Zr. A statistical analysis was performed using one-way analysis of variance and post hoc Tukey's HSD tests (α = 0.05). The experimental results indicate that the TP values and contrast ratio of the M-Zr samples were 14.85 and 0.83, respectively. Co−Cr had the highest ΔE (6.08) and lowest C value (7.52); PEKK had the lowest ΔE (2.60), and PEEK had the highest C value (12.23) (p < 0.05). Notably, the ΔE values of CR (3.13), PEEK (2.86), and PEKK were within clinical indicators (ΔE < 3.7). Based on these results, it can be concluded that the abutment material has a significant effect on the final color of the M-Zr, and PEEK or PEKK resulted in good color accuracy. When choosing the dental MR, traditional zirconia, or metals as abutment materials, colored or opaque cement might be required to eliminate color distortion and achieve desirable optical properties.