| 1. |
- Feng, Ruizhi, et al.
(författare)
-
Mutations in TUBB8 and Human Oocyte Meiotic Arrest.
- 2016
-
Ingår i: The New England journal of medicine. - 1533-4406. ; 374:3, s. 223-232
-
Tidskriftsartikel (refereegranskat)abstract
- Background Human reproduction depends on the fusion of a mature oocyte with a sperm cell to form a fertilized egg. The genetic events that lead to the arrest of human oocyte maturation are unknown. Methods We sequenced the exomes of five members of a four-generation family, three of whom had infertility due to oocyte meiosis I arrest. We performed Sanger sequencing of a candidate gene, TUBB8, in DNA samples from these members, additional family members, and members of 23 other affected families. The expression of TUBB8 and all other β-tubulin isotypes was assessed in human oocytes, early embryos, sperm cells, and several somatic tissues by means of a quantitative reverse-transcriptase-polymerase-chain-reaction assay. We evaluated the effect of the TUBB8 mutations on the assembly of the heterodimer consisting of one α-tubulin polypeptide and one β-tubulin polypeptide (α/β-tubulin heterodimer) in vitro, on microtubule architecture in HeLa cells, on microtubule dynamics in yeast cells, and on spindle assembly in mouse and human oocytes. Results We identified seven mutations in the primate-specific gene TUBB8 that were responsible for oocyte meiosis I arrest in 7 of the 24 families. TUBB8 expression is unique to oocytes and the early embryo, in which this gene accounts for almost all the expressed β-tubulin. The mutations affect chaperone-dependent folding and assembly of the α/β-tubulin heterodimer, disrupt microtubule behavior on expression in cultured cells, alter microtubule dynamics in vivo, and cause catastrophic spindle-assembly defects and maturation arrest on expression in mouse and human oocytes. Conclusions TUBB8 mutations have dominant-negative effects that disrupt microtubule behavior and oocyte meiotic spindle assembly and maturation, causing female infertility. (Funded by the National Basic Research Program of China and others.).
|
|
| 2. |
- Camacho, Rafael, et al.
(författare)
-
2D polarization imaging as a low-cost fluorescence method to detect α-synuclein aggregation ex vivo in models of Parkinson’s disease
- 2018
-
Ingår i: Communications Biology. - : Springer Science and Business Media LLC. - 2399-3642. ; 1
-
Tidskriftsartikel (refereegranskat)abstract
- A hallmark of Parkinson’s disease is the formation of large protein-rich aggregates in neurons, where α-synuclein is the most abundant protein. A standard approach to visualize aggregation is to fluorescently label the proteins of interest. Then, highly fluorescent regions are assumed to contain aggregated proteins. However, fluorescence brightness alone cannot discriminate micrometer-sized regions with high expression of non-aggregated proteins from regions where the proteins are aggregated on the molecular scale. Here, we demonstrate that 2-dimensional polarization imaging can discriminate between preformed non-aggregated and aggregated forms of α-synuclein, and detect increased aggregation in brain tissues of transgenic mice. This imaging method assesses homo-FRET between labels by measuring fluorescence polarization in excitation and emission simultaneously, which translates into higher contrast than fluorescence anisotropy imaging. Exploring earlier aggregation states of α-synuclein using such technically simple imaging method could lead to crucial improvements in our understanding of α-synuclein-mediated pathology in Parkinson’s Disease.
|
|
| 3. |
- Shi, Juanzi, et al.
(författare)
-
Energy transfer in multi-funnel systems quantitatively assessed by two-dimensional polarization imaging and single funnel approximation: From single molecules to ensembles
- 2022
-
Ingår i: The Journal of chemical physics. - : AIP Publishing. - 1089-7690 .- 0021-9606. ; 156:7
-
Tidskriftsartikel (refereegranskat)abstract
- Two-dimensional polarization imaging (2D POLIM) is an experimental method where correlations between fluorescence excitation- and fluorescence emission-polarization properties are measured. One way to analyze 2D POLIM data is to apply a so-called single funnel approximation (SFA). The SFA allows for quantitative assessment of energy transfer between chromophores with identical spectra [homo-FRET (Förster resonance energy transfer)]. In this paper, we run a series of computer experiments to investigate the applicability of the analysis based on the SFA to various systems ranging from single multichromophoric systems to isotropic ensembles. By setting various scenarios of energy transfer between individual chromophores within a single object, we were able to define the borders of the practical application of SFA. It allowed us to reach a more comprehensive interpretation of the experimental data in terms of uncovering the internal arrangement of chromophores in the system and energy transfer between them. We also found that the SFA can always formally explain the data for isotropic ensembles and derived a formula connecting the energy funneling efficiency parameter and traditional fluorescence anisotropy.
|
|
| 4. |
- Shi, Juanzi
(författare)
-
Nanoscale organization of luminescent materials and their polarization properties investigated by two-dimensional polarization imaging
- 2021
-
Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Semiconductor materials (e.g., conjugated polymers, metal halide perovskites) have been widely used in solar cells, light-emitting diodes, and photodetectors. Organic conjugated systems have high mechanical flexibility and low costs for production. Metal halide perovskites have the advantage of strong light absorption, long charge-carrier diffusion lengths, and low intrinsic surface recombination.Polarization-sensitive single-molecule methods have been extensively used to study the chromophore organization and excitation energy transfer (EET) process. Our novel polarization technique, two-dimensional polarization imaging (2D POLIM) is designed to simultaneously measure and control both the excitation and emission polarization characteristics of an individual object. A model based on single funnel approximation (SFA) is applied to fit the 2D polarization portrait obtained from 2D POLIM measurements. 2D POLIM in combination with the SFA model allows the quantitative characterization of EET efficiency. Overall, A large number of polarization parameters, e.g., modulation depths, phases, luminescence shift, fluorescence anisotropy, energy funneling efficiency, and properties of the EET-emitter, can be extracted from 2D polarization portraits. They give a full picture of chromophores’ organization and a quantitative measure of the EET process.In this thesis, we applied the 2D POLIM technique to investigate the fundamental optoelectronic process in different types of luminescent materials. H-aggregates forming in spin-cast conjugated films are visualized by modulation depth and phase imaging contrast. Light-harvesting efficiency shows the efficient ET within the amorphous phase and poor ET between H-aggregates due to the less overlap between absorption and emission spectra. Together with single-molecule spectroscopy and scanning electron microscope, we studied the polarization property of individual MAPbBr3 aggregates, which shows the well-known dielectric screening effect cannot fully explain the absorption polarization from weakly elongated objects (even with irregular shapes). We propose that power dependent quantum yield can further increase the modulation depth of excitation. 2D POLIM was also applied to explore the aggregation state of proteins in the biological system. Furthermore, we did a series of computer experiments to examine and improve the SFA model. We break the limit of energy funneling efficiency and propose an asymmetric three-dipole model, which is more applicable for multi-chromophore systems. In the future, quantitative phase-contrast imaging and time-resolved 2D POLIM might be further developed
|
|
| 5. |
- Shi, Juanzi, et al.
(författare)
-
Photo-Oxidation Reveals H-Aggregates Hidden in Spin-Cast-Conjugated Polymer Films as Observed by Two-Dimensional Polarization Imaging
- 2019
-
Ingår i: Chemistry of Materials. - : American Chemical Society (ACS). - 0897-4756 .- 1520-5002. ; 31:21, s. 8927-8936
-
Tidskriftsartikel (refereegranskat)abstract
- Spin-cast intermolecular interactions in conjugated polymer films lead to the formation of excited states delocalized over a few oriented and tightly packed conjugated segments. The optoelectronic properties of conjugated polymers are strongly dependent on the presence of such oriented domains at a nanoscale level. We observe oriented domains as large as several micrometers in size spontaneously formed in spin-cast PBDT-TPD films. Two-dimensional polarization imaging of fresh and photodegraded films showed a much higher visibility of the oriented domains in the degraded samples. We propose that the film is a mixture of two phases with different degrees of chain alignment. The photoluminescence of the more anisotropic phase is more stable against photodegradation in comparison with the less anisotropic phase. Photodegradation predominately quenches photoluminescence of the less anisotropic phase making the oriented domains more visible in the polarization contrasts. Spectral and energy transfer properties of the more oriented phase allowed us to assign it to weakly coupled H-aggregates with the suppressed 0-0 vibronic transition. Stable photoluminescence of H-aggregates in comparison with that of nonaggregated (less oriented) chains may help to understand degradation mechanisms of polymer devices and shows the role of energy transfer in this process. Selective degradation-induced quenching can reveal hidden inhomogeneity of conjugated polymer films.
|
|
| 6. |
- Shi, Juanzi, et al.
(författare)
-
Photoluminescence Polarization of MAPbBr3 Perovskite Nanostructures. Can the Dielectric Contrast Effect Explain It?
- 2022
-
Ingår i: ACS Photonics. - : American Chemical Society (ACS). - 2330-4022. ; 9:12, s. 3888-3898
-
Tidskriftsartikel (refereegranskat)abstract
- The dielectric contrast effect is usually evoked to explain anisotropy of optical properties of elongated nanoobjects, for example, semiconductor nanowires. We applied two-dimensional polarization imaging microscopy to measure the polarization of photoluminescence (PL) excitation and PL intensity of nanoaggregates of in-situ formed MAPbBr3perovskite nanoparticles in a stretched polymeric matrix. Scanning electron microscopy images of these objects were also acquired to characterize their sizes and shapes. We find that individual perovskite aggregates with sizes of 100-300 nm often possess a PL excitation polarization degree as high as 0.5-0.9, which is up to three times higher than the polarization degree of absorption predicted by the dielectric contrast effect. Small aggregates of nanoparticles possess an emission polarization degree substantially higher than that of excitation. Computer simulations of many possible scenarios show that the dielectric contrast alone cannot quantitatively explain the polarization properties of the studied objects. We propose energy transfer to localized emitting sites and the dependence of PL yield on excitation power density as possible factors strongly influencing the polarization properties of PL emission and PL excitation, respectively.
|
|
