Investigation of the TeO2/GeO2 Ratio on the Spectroscopic Properties of Eu3+-Doped Oxide Glasses for Optical Fiber Application

This study presented an analysis of the TeO₂/GeO₂ molar ratio in an oxide glass system. A family of melt-quenched glasses with the range of 0–35 mol% of GeO₂ has been characterized by using DSC, Raman, MIR, refractive index, PLE, PL spectra, and time-resolved spectral measurements. The increase in the content of germanium oxide caused an increase in the transition temperature but a decrease in the refractive index. The photoluminescence spectra of europium ions were examined under the excitation of 465 nm, corresponding to ⁷F₀ → ⁵D₂ transition. The PSB (phonon sidebands) analysis was carried out to determine the phonon energy of the glass hosts. It was reported that the red (⁵D₀ → ⁷F₂) to orange (⁵D₀ → ⁷F₁) fluorescence intensity ratio for Eu³⁺ ions decreased from 4.49 (Te0Ge) to 3.33 (Te15Ge) and showed a constant increase from 4.58 (Te20Ge) to 4.88 (Te35Ge). These optical features were explained in structural studies, especially changes in the coordination of ^[4]Ge to ^[6]Ge. The most extended lifetime was reported for the Eu³⁺ doped glass with the highest content of GeO₂. This glass was successfully used for the drawing of optical fiber.     

Near-IR Luminescence of Rare-Earth Ions (Er3+, Pr3+, Ho3+, Tm3+) in Titanate–Germanate Glasses under Excitation of Yb3+

Inorganic glasses co-doped with rare-earth ions have a key potential application value in the field of optical communications. In this paper, we have fabricated and then characterized multicomponent TiO₂-modified germanate glasses co-doped with Yb³⁺/Ln³⁺ (Ln = Pr, Er, Tm, Ho) with excellent spectroscopic properties. Glass systems were directly excited at 980 nm (the ²F_7/2 → ²F_5/2 transition of Yb³⁺). We demonstrated that the introduction of TiO₂ is a promising option to significantly enhance the main near-infrared luminescence bands located at the optical telecommunication window at 1.3 μm (Pr³⁺: ¹G₄ → ³H₅), 1.5 μm (Er³⁺: ⁴I_13/2 → ⁴I_15/2), 1.8 μm (Tm³⁺: ³F₄ → ³H₆) and 2.0 μm (Ho³⁺: ⁵I₇ → ⁷I₈). Based on the lifetime values, the energy transfer efficiencies (η_ET) were estimated. The values of η_ET are changed from 31% for Yb³⁺/Ho³⁺ glass to nearly 53% for Yb³⁺/Pr³⁺ glass. The investigations show that obtained titanate–germanate glass is an interesting type of special glasses integrating luminescence properties and spectroscopic parameters, which may be a promising candidate for application in laser sources emitting radiation and broadband tunable amplifiers operating in the near-infrared range.     

Nonstoichiometric LaO0.65F1.7 Structure and Its Green Luminescence Property Doped with Bi3+ and Tb3+ Ions for Applying White UV LEDs

Red–green–blue phosphors excited by ultraviolet (UV) radiation for white light LEDs have received much attention to improve the efficiency, color rendering index (CRI), and chromatic stability. The spectral conversion of a rare-earth ion-doped nonstoichiometric LaO_0.65F_1.7 host was explored with structural analysis in this report. The nonstoichiometric structure of a LaO_0.65F_1.7 compound, synthesized by a solid-state reaction using La₂O₃ and excess NH₄F precursors, was analyzed by synchrotron X-ray powder diffraction. The crystallized LaO_0.65F_1.7 host, which had a tetragonal space group of P4/nmm, contained 9- and 10-coordinated La³⁺ sites. Optical materials composed of La_1−p−qBi_pTb_qO_0.65F_1.7 (p = 0 and 0.01; q = 0–0.2) were prepared at 1050 °C for 2 h, and the single phase of the obtained phosphors was indexed by X-ray diffraction analysis. The photoluminescence spectra of the energy transfer from Bi³⁺ to Tb³⁺ were obtained upon excitation at 286 nm in the nonstoichiometric host lattice. The desired Commission Internationale de l’Eclairage (CIE) values of the phosphors were calculated. The intense green La_0.89Bi_0.01Tb_0.1O_0.65F_1.7 phosphor with blue and red optical materials was fabricated on a 275 nm UV-LED chip, resulting in white light, and the internal quantum efficiency, CRI, correlated color temperature, and CIE of the pc LED were characterized.     

Combustion Synthesis and Photoluminescence Properties of Red-Emitting CaAlSiN3:Eu2+ Phosphor for White-LEDs

A combustion synthesis method has been developed for synthesis of Eu²⁺ doped CaAlSiN₃ phosphor and its photoluminescence properties were investigated. Ca, Al, Si, and Eu₂O₃ powders were used as the Ca, Al, Si and Eu sources. The addition of NaN₃, NH₄Cl and Si₃N₄ powders was found to increase significantly the product yield. These powders were mixed and pressed into a compact, which was then wrapped up with an igniting agent (i.e., Mg+Fe₃O₄). The compact was ignited by electrical heating under a N₂ pressure of ≤1.0 MPa. Effects of these experimental parameters on the product yield were investigated and a reaction mechanism was proposed. The synthesized CaAlSiN₃:Eu²⁺ phosphor absorbs light in the region of 200–600 nm and shows a broad band emission in the region of 500–800 nm due to the 4f⁶5d¹ → 4f⁷ transition of Eu²⁺. The sample doped with Eu²⁺ at the optimized molar ratio of 0.04 is efficiently excited by the blue light (460 nm) and generates emission peaking at ~650 nm with peak emission intensity ~106% of a commercially available phosphor, YAG:Ce³⁺(P46-Y3).The internal quantum efficiency of the synthesized phosphor was measured to be 71%, compared to 69% of the YAG:Ce³⁺ (P46-Y3).     

Structure and Photoluminescence Properties of Dy3+ Doped Phosphor with Whitlockite Structure

Whitlockite has the advantages of a low sintering temperature, high stability, and a low fabrication cost, and it is widely used as the host for luminescent material. In this study, Ca_1.8Li_0.6La_0.6−x(PO₄)₂:xDy³⁺ phosphor was prepared by the high-temperature solid-state method, and its structure, composition, and luminescence properties were systematically studied. The results showed that a new whitlockite type matrix was prepared by replacing Ca²⁺ in whitlockite with monovalent and trivalent cations. The prepared phosphors belonged to a hexagonal crystal system with a particle size in the range of 5–20 μm. Under the excitation of 350 nm UV light, the samples emitted white light, and there were mainly two stronger emission peaks at 481 nm in the blue band and 573 nm in the yellow band, which correspond to the electron transitions at ⁴F_9/2→⁶H_15/2 and ⁴F_9/2→⁶H_13/2 of Dy³⁺, respectively. The optimal doping concentration of Dy³⁺ in Ca_1.8Li_0.6La_0.6(PO₄)₂ matrix was 0.03 (mol%). The main mechanism of concentration quenching in the sample was dipole–dipole energy transfer. When the temperature was 130 °C, the luminescence intensity of the samples was 78.7% of that at 30 °C, and their thermal quenching activation energy was 0.25 eV. The CIE coordinates of the sample at 30 °C were (0.2750, 0.3006), and their luminescent colors do not change with temperature. All the results indicate that Ca_1.8Li_0.6La_0.6−x(PO₄)₂:xDy³⁺ phosphor is a luminescent material with good luminescence performance and thermal stability, which shows a promising application in the field of LED display.     

Wide Concentration Range of Tb3+ Doping Influence on Scintillation Properties of (Ce,Tb, Gd)3Ga2Al3O12 Crystals Grown by the Optical Floating Zone Method

To obtain a deeper understand of the energy transfer mechanism between Ce³⁺ and Tb³⁺ ions in the aluminum garnet hosts, (Ce, Tb, Gd)₃Ga₂Al₃O₁₂ (GGAG:Ce, Tb) single crystals grown by the optical floating zone (OFZ) method were investigated systematically in a wide range of Tb³⁺ doping concentration (1–66 at.%). Among those, crystal with 7 at.% Tb reached a single garnet phase while the crystals with other Tb³⁺ concentrations are mixed phases of garnet and perovskite. Obvious Ce and Ga loss can be observed by an energy dispersive X-ray spectroscope (EDS) technology. The absorption bands belonging to both Ce³⁺ and Tb³⁺ ions can be observed in all crystals. Photoluminescence (PL) spectra show the presence of an efficient energy transfer from the Tb³⁺ to Ce³⁺ and the gradually quenching effect with increasing of Tb³⁺ concentration. GGAG: 1% Ce³⁺, 7% Tb³⁺ crystal was found to possess the highest PL intensity under excitation of 450 nm. The maximum light yield (LY) reaches 18,941 pho/MeV. The improved luminescent and scintillation characteristics indicate that the cation engineering of Tb³⁺ can optimize the photoconversion performance of GGAG:Ce.     

Red-Emitting Hybrid Based on Eu3+-dbm Complex Anchored on Silica Nanoparticles Surface by Carboxylic Acid for Biomarker Application

Luminescent organic-inorganic hybrids containing lanthanides (Ln³⁺) have been prominent for applications such as luminescent bio-probes in biological assays. In this sense, a luminescent hybrid based on dense silica (SiO₂) nanospheres decorated with Eu³⁺ β–diketonate complexes using dibenzoylmethane (Hdbm) as a luminescent antenna was developed by using a hierarchical organization in four steps: (i) anchoring of 3-aminopropyltriethoxysilane (APTES) organosilane on the SiO₂ surface, (ii) formation of a carboxylic acid ligand, (iii) coordination of Eu³⁺ to the carboxylate groups and (iv) coordination of dbm⁻ to Eu³⁺. The hybrid structure was elucidated through the correlation of thermogravimetry, silicon nuclear magnetic resonance and photoluminescence. Results indicate that the carboxylic acid-Eu³⁺-dbm hybrid was formed on the surface of the particles with no detectable changes on their size or shape after all the four steps (average size of 32 ± 7 nm). A surface charge of −27.8 mV was achieved for the hybrid, assuring a stable suspension in aqueous media. The Eu³⁺ complex provides intense red luminescence, characteristic of Eu^{3+ 5}D₀→⁷F_J electronic transitions, with an intrinsic emission quantum yield of 38%, even in an aqueous suspension. Therefore, the correlation of luminescence, structure, particle morphology and fluorescence microscopy images make the hybrid promising for application in bioimaging.     

Crystallization Mechanism and Optical Properties of Antimony-Germanate-Silicate Glass-Ceramic Doped with Europium Ions

Glass-ceramic is semi-novel material with many applications, but it is still problematic in obtaining fibers. This paper aims to develop a new glass-ceramic material that is a compromise between crystallization, thermal stability, and optical properties required for optical fiber technology. This compromise is made possible by an alternative method with a controlled crystallization process and a suitable choice of the chemical composition of the core material. In this way, the annealing process is eliminated, and the core material adopts a glass-ceramic character with high transparency directly in the drawing process. In the experiment, low phonon antimony-germanate-silicate glass (SGS) doped with Eu³⁺ ions and different concentrations of P₂O₅ were fabricated. The glass material crystallized during the cooling process under conditions similar to the drawing processes’. Thermal stability (DSC), X-ray photo analysis (XRD), and spectroscopic were measured. Eu³⁺ ions were used as spectral probes to determine the effect of P₂O₅ on the asymmetry ratio for the selected transitions (⁵D₀ → ⁷F₁ and ⁵D₀ → ⁷F₂). From the measurements, it was observed that the material produced exhibited amorphous or glass-ceramic properties, strongly dependent on the nucleator concentration. In addition, the conducted study confirmed that europium ions co-form the EuPO₄ structure during the cooling process from 730 °C to room temperature. Moreover, the asymmetry ratio was changed from over 4 to under 1. The result obtained confirms that the developed material has properties typical of transparent glass-ceramic while maintaining high thermal stability, which will enable the fabrication of fibers with the glass-ceramic core.     

Spectroscopic Properties of Inorganic Glasses Doped with Pr3+: A Comparative Study

The results presented in this communication concern visible and near-IR emission of Pr³⁺ ions in selected inorganic glasses, i.e., borate-based glass with Ga₂O₃ and BaO, lead-phosphate glass with Ga₂O₃, gallo-germanate glass modified by BaO/BaF₂, and multicomponent fluoride glass based on InF₃. Glasses present several emission bands at blue, reddish orange, and near-infrared spectral ranges, which correspond to 4f–4f electronic transitions of Pr³⁺. The profiles of emission bands and their relative intensity ratios depend strongly on glass-host. Visible emission of Pr³⁺ ions is tuned from red/orange for borate-based glass to nearly white light for multicomponent fluoride glass based on InF₃. The positions and spectral linewidths for near-infrared luminescence bands at the optical telecommunication window corresponding to the ¹G₄ → ³H₅, ¹D₂ → ¹G₄, and ³H₄ → ³F₃,³F₄ transitions of Pr³⁺ are dependent on glass-host matrices and excitation wavelengths. Low-phonon fluoride glasses based on InF₃ and gallo-germanate glasses with BaO/BaF₂ are excellent candidates for broadband near-infrared optical amplifiers. Spectroscopic properties of Pr³⁺-doped glasses are compared and discussed in relation to potential optical applications.     

Synthesis,Characterization and Biodistribution of GdF3:Tb3+@RB Nanocomposites

Herein we report the development of a nanocomposite for X-ray-induced photodynamic therapy (X-PDT) and computed tomography (CT) based on PEG-capped GdF₃:Tb³⁺ scintillating nanoparticles conjugated with Rose Bengal photosensitizer via electrostatic interactions. Scintillating GdF₃:Tb³⁺ nanoparticles were synthesized by a facile and cost-effective wet chemical precipitation method. All synthesized nanoparticles had an elongated “spindle-like” clustered morphology with an orthorhombic structure. The structure, particle size, and morphology were determined by transmission electron microscopy (TEM), X-ray diffraction (XRD), and dynamic light scattering (DLS) analysis. The presence of a polyethylene glycol (PEG) coating and Rose Bengal conjugates was proved by Fourier-transform infrared spectroscopy (FTIR), thermogravimetric analysis (TG), and ultraviolet–visible (UV-vis) analysis. Upon X-ray irradiation of the colloidal PEG-capped GdF₃:Tb³⁺–Rose Bengal nanocomposite solution, an efficient fluorescent resonant energy transfer between scintillating nanoparticles and Rose Bengal was detected. The biodistribution of the synthesized nanoparticles in mice after intravenous administration was studied by in vivo CT imaging.     

Synthesis and Characterization of Li2MgGeO4:Ho3+

In this work, the synthesis and characterization of Li₂MgGeO₄:Ho³⁺ ceramics were reported. The X-ray diffraction measurements revealed that the studied ceramics belong to the monoclinic Li₂MgGeO₄. Luminescence properties were analyzed in the visible spectral range. Green and red emission bands correspondent to the ⁵F₄,⁵S₂→⁵I₈ and ⁵F₅→⁵I₈ transitions of Ho³⁺ were observed, and their intensities were significantly dependent on activator concentration. Luminescence spectra were also measured under direct excitation of holmium ions or ceramic matrix. Holmium ions were inserted in crystal lattice Li₂MgGeO₄, giving broad blue emission and characteristic 4f-4f luminescent transitions of rare earths under the selective excitation of the ceramic matrix. The presence of the energy transfer process between the host lattice and Ho³⁺ ions was suggested.     

1D-Zigzag Eu3+/Tb3+ Coordination Chains as Luminescent Ratiometric Thermometers Endowed with Multicolor Emission

Two homometallic Coordination Polymers (CPs) with composition [Ln(hfac)₃bipy]_n (Ln³⁺ = Eu³⁺, 1, and Tb³⁺, 2; hfac = hexafluoroacetylacetonato, bipy = 4,4′-bipyridine) were used to develop a family of ratiometric luminescent thermometers containing Eu³⁺ and Tb³⁺ as red and green emitters, respectively. The thermometric properties of pure CPs and of their mixtures having an Eu³⁺/Tb³⁺ molar ratio of 1:1, 1:3, 1:5, and 1:10 (samples: Eu1Tb1, Eu1Tb3, Eu1Tb5, and Eu1Tb10) were studied in the 83–383 K temperature range. Irrespective of the chemical composition, we observed similar thermometric responses characterized by broad applicative temperature ranges (from 100 to 165 K wide), and high relative thermal sensitivity values (S_r), up to 2.40% K⁻¹, in the physiological temperature range (298–318 K). All samples showed emissions endowed with peculiar and continuous color variation from green (83 K) to red (383 K) that can be exploited to develop a colorimetric temperature indicator. At fixed temperature, the color of the emitted light can be tuned by varying composition and excitation wavelength.     

Thermal Evolutions to Glass-Ceramics Bearing Calcium Tungstate Crystals in Borate Glasses Doped with Photoluminescent Eu3+ Ions

Thermal evolutions of calcium-tungstate-borate glasses were investigated for the development of luminescent glass-ceramics by using Eu³⁺ dopant in a borate glass matrix with calcium tungstate, which was expected to have a combined character of glass and ceramics. This study revealed that single-phase precipitation of CaWO₄ crystals in borate glass matrix was possible by heat-treatment at a temperature higher than glass transition temperature T_g for (100−x) (33CaO-67B₂O₃)−xCa₃WO₆ (x = 8−15 mol%). Additionally, the crystallization of CaWO₄ was found by Raman spectroscopy due to the formation of W=O double bondings of WO₄ tetrahedra in the pristine glass despite starting with the higher calcium content of Ca₃WO₆. Eu³⁺ ions were excluded from the CaWO₄ crystals and positioned in the borate glass phase as a stable site for them, which provided local environments in higher symmetry around Eu³⁺ ions.     

The Upconversion Luminescence of Er3+/Yb3+/Nd3+ Triply-Doped β-NaYF4 Nanocrystals under 808-nm Excitation

In this paper, Nd³⁺–Yb³⁺–Er³⁺-doped β-NaYF₄ nanocrystals with different Nd³⁺ concentrations are synthesized, and the luminescence properties of the upconversion nanoparticles (UCNPs) have been studied under 808-nm excitation for sensitive biological applications. The upconversion luminescence spectra of NaYF₄ nanoparticles with different dopants under 808-nm excitation proves that the Nd³⁺ ion can absorb the photons effectively, and the Yb³⁺ ion can play the role of an energy-transfer bridging ion between the Nd³⁺ ion and Er³⁺ ion. To investigate the effect of the Nd³⁺ ion, the decay curves of the ⁴S_3/2 → ⁴I_15/2 transition at 540 nm are measured and analyzed. The NaYF₄: 20% Yb³⁺, 2% Er³⁺, 0.5% Nd³⁺ nanocrystals have the highest emission intensity among all samples under 808-nm excitation. The UC (upconversion) mechanism under 808-nm excitation is discussed in terms of the experimental results.     

Tunable Luminescence and Energy Transfer of Sr3B2O6:Ce3+, Sm3+ Phosphors with Potential Anti-Counterfeiting Applications

Sm³⁺ and Ce³⁺ singly doped and Sm³⁺ and Ce³⁺ co-doped Sr₃B₂O₆ phosphors are prepared via a high-temperature solid-state reaction method. The crystal structure and phase purity are characterized by X-ray diffraction (XRD) analyses. The Sm³⁺-doped sample displays an emission in the orange-red region, with the strongest emission line at about 648 nm and possessing a good luminescence thermal stability between 78 and 500 K. With the increase in the Sm³⁺ content, the concentration quenching is observed due to the cross-relaxation (CR) processes among the Sm³⁺ ions. Upon 340 nm excitation, the Ce³⁺-doped phosphor presents a broad emission band in the blue region with a maximum at about 420 nm, which overlaps well with the ⁶H_5/2 → ⁶P_3/2 excitation line of Sm³⁺ and implies the possible energy transfer from Ce³⁺ to Sm³⁺. The spectral and decay measurements of the Ce³⁺ and Sm³⁺ co-doped samples are conducted and the Inokuti–Hirayama (I-H) model is adopted to analyze the luminescence decay dynamics of the donor Ce³⁺. Owing to the evident sensitization of the Sm³⁺ by the Ce³⁺ ions, the co-doped samples exhibit color variation under different wavelength excitations, endowing them with potential applications in optical anti-counterfeiting.     

Eu3+ as a Powerful Structural and Spectroscopic Tool for Glass Photonics

The unique properties of the Eu³⁺ ion make it a powerful spectroscopic tool to investigate structure or follow processes and mechanisms in several high-tech application areas such as biology and health, structural engineering, environment monitoring systems and quantum technology, mainly concerning photonics. The traditional method is to exploit the unique photoluminescent properties of Eu³⁺ ions to understand complex dynamical processes and obtain information useful to develop materials with specific characteristics. The objective of this review is to focus on the use of Eu³⁺ optical spectroscopy in some condensed matter issues. After a short presentation of the more significant properties of the Eu³⁺ ion, some examples regarding its use as a probe of the local structure in sol–gel systems are presented. Another section is devoted to dynamical processes such as the important technological role of nanocrystals as rare-earth sensitizers. The appealing effect of the site-selection memory, observed when exciting different sites into the ⁵D₁ state, which the ⁵D₀ → ⁷F₀ emission band reflects following the sites’ distribution, is also mentioned. Finally, a section is devoted to the use of Eu³⁺ in the development of a rare-earth-based platform for quantum technologies.     

Micropowder Ca2YMgScSi3O12:Ce Silicate Garnet as an Efficient Light Converter for White LEDs

This work is dedicated to the crystallization and luminescent properties of a prospective Ca₂YMgScSi₃O₁₂:Ce (CYMSSG:Ce) micropowder (MP) phosphor converter (pc) for a white light–emitting LED (WLED). The set of MP samples was obtained by conventional solid-phase synthesis using different amounts of B₂O₃ flux in the 1–5 mole percentage range. The luminescent properties of the CYMSSG:Ce MPs were investigated at different Ce³⁺ concentrations in the 1–5 atomic percentage range. The formation of several Ce³⁺ multicenters in the CYMSSG:Ce MPs was detected in the emission and excitation spectra as well as the decay kinetics of the Ce³⁺ luminescence. The creation of the Ce³⁺ multicenters in CYMSSG:Ce garnet results from: (i) the substitution by the Ce³⁺ ions of the heterovalent Ca²⁺ and Y³⁺ cations in the dodecahedral position of the garnet host; (ii) the inhomogeneous local environment of the Ce³⁺ ions when the octahedral positions of the garnet are replaced by heterovalent Mg²⁺ and Sc³⁺ cations and the tetrahedral positions are replaced by Si⁴⁺ cations. The presence of Ce³⁺ multicenters significantly enhances the Ce³⁺ emission band in the red range in comparison with conventional YAG:Ce phosphor. Prototypes of the WLEDs were also created in this work by using CYMSSG:Ce MP films as phosphor converters. Furthermore, the dependence of the photoconversion properties on the layer thickness of the CYMSSG:Ce MP was studied as well. The changes in the MP layer thickness enable the tuning of the white light thons from cold white/daylight to neutral white. The obtained results are encouraging and can be useful for the development of a novel generation of pcs for WLEDs.     

Structural and Luminescence Behavior of Nanocrystalline Orthophosphate KMeY(PO4)2: Eu3+ (Me = Ca,Sr) Synthesized by Hydrothermal Method

KMeY(PO₄)₂:5% Eu³⁺ phosphates have been synthesized by a novel hydrothermal method. Spectroscopic, structural, and morphological properties of the obtained samples were investigated by X-ray, TEM, Raman, infrared, absorption, and luminescence studies. The microscopic analysis of the obtained samples showed that the mean diameter of synthesized crystals was about 15 nm. The KCaY(PO₄)₂ and KSrY(PO₄)₂ compounds were isostructural and they crystallized in a rhabdophane-type hexagonal structure with the unit-cell parameters a = b ≈ 6.90 Å, c ≈ 6.34 Å, and a = b ≈ 7.00 Å, c ≈ 6.42 Å for the Ca and Sr compound, respectively. Spectroscopic investigations showed intense ⁵D₀ → ⁷F₄ transitions connected with D₂ site symmetry of Eu³⁺ ions. Furthermore, for the sample annealed at 500 °C, europium ions were located in two optical sites, on the surface of grains and in the bulk. Thermal treatment of powders at high temperature provided better grain crystallinity and only one position of dopant in the crystalline structure. The most intense emission was possessed by the KSrY(PO₄)₂:5% Eu³⁺ sample calcinated at 500 °C.     

Regulation of longevity by depolarization-induced activation of PLC-β–IP3R signaling in neurons

Mitochondrial ATP production is a well-known regulator of neuronal excitability. The reciprocal influence of plasma-membrane potential on ATP production, however, remains poorly understood. Here, we describe a mechanism by which depolarized neurons elevate the somatic ATP/ADP ratio in Drosophila glutamatergic neurons. We show that depolarization increased phospholipase-Cβ (PLC-β) activity by promoting the association of the enzyme with its phosphoinositide substrate. Augmented PLC-β activity led to greater release of endoplasmic reticulum Ca²⁺ via the inositol trisphosphate receptor (IP₃R), increased mitochondrial Ca²⁺ uptake, and promoted ATP synthesis. Perturbations that decoupled membrane potential from this mode of ATP synthesis led to untrammeled PLC-β–IP₃R activation and a dramatic shortening of Drosophila lifespan. Upon investigating the underlying mechanisms, we found that increased sequestration of Ca²⁺ into endolysosomes was an intermediary in the regulation of lifespan by IP₃Rs. Manipulations that either lowered PLC-β/IP₃R abundance or attenuated endolysosomal Ca²⁺ overload restored animal longevity. Collectively, our findings demonstrate that depolarization-dependent regulation of PLC-β–IP₃R signaling is required for modulation of the ATP/ADP ratio in healthy glutamatergic neurons, whereas hyperactivation of this axis in chronically depolarized glutamatergic neurons shortens animal lifespan by promoting endolysosomal Ca²⁺ overload.

Spatially circumscribed ATP production at nerve termini is predicated on local mitochondria that are energized when voltage-gated Ca²⁺ channels provide the [Ca²⁺] elevations needed to overcome the low sensitivity of the mitochondrial Ca²⁺ uniporter (MCU) (1–3). In neuronal soma, however, bulk cytosolic [Ca²⁺] is not elevated to levels needed for mitochondrial sequestration. Rather, mitochondrial Ca²⁺ uptake in the somatodendritic compartment occurs at specialized points of contact between mitochondria and endoplasmic reticulum (ER) where Ca²⁺ released by IP₃Rs is transferred into the mitochondrial matrix (4). Approximately 75 to 90% of the somatic ATP synthesized following interorganellar transfer of Ca²⁺ is consumed by Na⁺/K⁺ ATPases, which help establish resting membrane potential and permit repolarization during activity (5, 6). Therefore, defects in neuronal ATP synthesis result in loss of membrane potential and hyperexcitability (6).Whether excitability of the somatic plasma membrane (PM) exerts reciprocal influence on mitochondrial [Ca²⁺] and ATP production remains poorly understood. In an attempt to fill some of the gaps in knowledge, we examined the effects of PM potential on mitochondrial ATP production and Ca²⁺ homeostasis in Drosophila neurons. Owing to recent reports of neuronal hyperexcitability being a driver of diminished longevity in organisms ranging from Caenorhabditis elegans to humans (7–9), we hoped our studies would inform insights into the regulation of aging and lifespan. Moreover, since neuronal hyperexcitability, Ca²⁺ dyshomeostasis, and bioenergetic dysfunction characterize neurodegenerative diseases (6, 10, 11), uncovering actionable molecular targets that bridge these perturbations may bear therapeutic value. Our findings reveal a previously unknown mechanism by which excitability regulates bioenergetics and Ca²⁺ signaling and points to the utility of this signaling circuit in the regulation of longevity.     

Multiple deprotonation paths of the nucleophile 3′-OH in the DNA synthesis reaction

DNA synthesis by polymerases is essential for life. Deprotonation of the nucleophile 3′-OH is thought to be the obligatory first step in the DNA synthesis reaction. We have examined each entity surrounding the nucleophile 3′-OH in the reaction catalyzed by human DNA polymerase (Pol) η and delineated the deprotonation process by combining mutagenesis with steady-state kinetics, high-resolution structures of in crystallo reactions, and molecular dynamics simulations. The conserved S113 residue, which forms a hydrogen bond with the primer 3′-OH in the ground state, stabilizes the primer end in the active site. Mutation of S113 to alanine destabilizes primer binding and reduces the catalytic efficiency. Displacement of a water molecule that is hydrogen bonded to the 3′-OH using the 2′-OH of a ribonucleotide or 2′-F has little effect on catalysis. Moreover, combining the S113A mutation with 2′-F replacement, which removes two potential hydrogen acceptors of the 3′-OH, does not reduce the catalytic efficiency. We conclude that the proton can leave the O3′ via alternative paths, supporting the hypothesis that binding of the third Mg²⁺ initiates the reaction by breaking the α–β phosphodiester bond of an incoming deoxyribonucleoside triphosphate (dNTP).

DNA polymerases catalyze the incorporation of deoxyribonucleoside monophosphate (dNMP) into an existing primer after binding a deoxyribonucleoside triphosphate (dNTP) complementary to a templating base. The catalytic process is thought to begin with deprotonation of the primer 3′-OH by a general base, continue by nucleophilic attack of the α-phosphate of the dNTP leading to a new phosphodiester bond, and finish with release of pyrophosphate after its protonation by a general acid (Fig. 1A) (1). This mechanism has long been established to require two Mg²⁺ ions (A and B) (2). However, visualizing the reaction intermediates using time-resolved X-ray crystallography reveals little movement of the protein but a third and transiently bound Mg²⁺ ion necessary for the DNA synthesis reaction (3, 4). Whether the reaction is driven by the third Mg²⁺ ion or the nucleophile 3′-OH, the identity of a general base responsible for deprotonating the primer 3′-OH remains unclear.Open in a separate windowFig. 1.DNA synthesis reaction. (A) Deprotonation and activation of the primer-end 3′-OH for the nucleophilic attack is hypothesized to be the first step. (B) In the three–Mg²⁺-ion catalysis, the first step appears to break the phosphodiester bond between the α- and β-phosphates of the dNTP. (C) Structural superposition of the reaction-ready (RS) and product state of the WT enzyme in extending the dT primer. The two Mg²⁺ ions (A and B sites) are bound and the 3′-OH is hydrogen-bonded to the transient WatN, which in turn is linked to the bulk solvent via another water molecule. The third Mg²⁺ (C site) is observed only with the products. (D) In the ground state with one Ca²⁺ (green sphere), the 3′-OH is hydrogen-bonded with S113 and not aligned for the inline nucleophilic attack in the absence of the A-site Mg²⁺. The aligned 3′-OH in the RS is shown as semitransparent sticks as a reference. (E) The transient WatN (circled in orange dashes) appears only in the RS and would clash with the C2′ in the GS (wheat) and PS (blue) as indicated by double arrowheads. (F) Sequence alignment of active-site residues around S113 (highlighted in red) of human Pol η, Saccharomyces cerevisiae Pol η (yeast), human Pol ι, human Pol κ, S. cerevisiae Rev1, Escherichia coli Pol IV, and Sulfolobus solfataricus Dpo4. The conserved secondary structures are shown above the sequences.Crystal structures of various DNA polymerases bound to DNA and dNTP, known as ternary complexes, reveal a conserved catalytic center with two Mg²⁺ ions coordinated by the conserved catalytic carboxylates, three phosphates of an incoming dNTP, and the primer 3′-OH (Fig. 1C) (5, 6). DNA polymerase has been likened to a right hand with the palm domain containing the catalytic residues, the finger domain closing on top of the nascent or replicating base pair, and the thumb domain binding the upstream DNA (product) duplex (7, 8). Many DNA polymerases undergo a large conformational change involving closing of the finger domain upon binding a correct dNTP (8), which was proposed to be a rate-limiting step (9). However, the rate of finger domain closing is much faster than the rate of chemical reaction (10–13). Moreover, the finger domain of the Y-family DNA polymerases is already closed even in the absence of an incoming nucleotide (14). Although intradomain motions have been implicated in one example (15), the rate-limiting step is probably the chemical reaction (3, 4, 16, 17).If nucleophilic attack is the first step, deprotonation of the primer 3′-OH by a general base would be essential to initiate the reaction (Fig. 1A). In the more than three decades since the first DNA polymerase structure was determined (7), no residues other than the conserved carboxylates that coordinate the two Mg²⁺ ions have been found to eliminate the catalytic activity when mutated. Computationally, the conserved carboxylates (1, 18) and the incoming nucleotide together with the water molecule bound to the A-site Mg²⁺ (WatA) have each been suggested to deprotonate the 3′-OH (12, 19–21). As the carboxylates and dNTPs are necessary for Mg²⁺ binding and the synthesis reaction, their role in deprotonation is nearly impossible to be experimentally tested.A third Mg²⁺ ion (occupying the C site) has been observed to transiently bind to dNTP in the reactions catalyzed by different DNA polymerases (3, 22–24). Unlike the A- and B-site Mg²⁺ ions, which are coordinated by the catalytic carboxylates, the C-site Mg²⁺ does not contact the enzyme at all. Its low affinity (k_d) matches the minimal Mg²⁺ concentration required for catalysis (3, 4). When two canonical Mg²⁺ ions are bound and reactants are aligned for the inline nucleophilic attack, the third Mg²⁺ ion does not bind readily (3, 4). Only when products are formed is the third Mg²⁺ ion observed to bind between the product DNA and pyrophosphate with four additional water ligands. Yet without the third Mg²⁺, no product can form (4). Its binding is thermal energy (temperature)-dependent, and concurrent with the product formation. The third Mg²⁺ ion may drive dNMP incorporation by breaking the α–β phosphodiester bond of dNTP and pushing the α-phosphate toward the 3′-OH for the new bond formation (4) (Fig. 1B). With the C-site Mg²⁺, deprotonating the 3′-OH is likely favored and does not need a strong general base.The in crystallo analysis of the DNA synthesis reaction catalyzed by human polymerase (Pol) η reveals three reaction states and two potential candidates to deprotonate the nucleophile. Initial dNTP binding with one divalent cation only (B site) leads to the ground state (GS) (Fig. 1D). Binding of the second Mg²⁺ converts the GS to the reactive state (RS), in which the 3′-OH is aligned for the inline nucleophilic attack, and appearance of the third Mg²⁺ (C site) is coupled with the product formation (product state; PS) (3, 4) (Fig. 1 C and E). In the GS, the 3′-OH is within 2.7 Å and hydrogen-bonded to the hydroxyl group of S113 (3). Although S113 is highly conserved among the Y-family polymerases (Fig. 1F) and may facilitate deprotonation of the 3′-OH, removal of the hydroxyl group by mutating S113 to Ala in human Pol η reduces the catalytic efficiency but does not eliminate catalysis (3). In the RS, the 3′-OH is 2.7 Å from a water molecule, which is termed WatN (N for nucleophile) and hypothesized to shuttle the proton off the 3′-OH to the bulk solvent (3). However, preliminary kinetic analysis showed that displacement of the water molecule by the 2′-OH of a ribonucleotide at the primer end leaves catalysis of DNA synthesis unaltered (3).Here we investigate in detail how the 3′-OH is deprotonated in the reaction catalyzed by human Pol η. Combining S113A mutant Pol η (S113A) and modified nucleotides at the primer 3′ end, we measured how the altered environment of the 3′-OH perturbed the steady-state kinetics and reaction process in crystallo. In addition, static structures and dynamics simulation are employed to explain observed kinetic parameters. In contrast to the hypothesis of a specific general base, we find that the proton of the 3′-OH can depart via multiple paths.