Recent studies have demonstrated the generation of midbrain-like organoids (MOs) from human pluripotent stem cells. However, the low efficiency of MO generation and the relatively immature and ...heterogeneous structures of the MOs hinder the translation of these organoids from the bench to the clinic. Here we describe the robust generation of MOs with homogeneous distribution of midbrain dopaminergic (mDA) neurons. Our MOs contain not only mDA neurons but also other neuronal subtypes as well as functional glial cells, including astrocytes and oligodendrocytes. Furthermore, our MOs exhibit mDA neuron-specific cell death upon treatment with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine, indicating that MOs could be a proper human model system for studying the in vivo pathology of Parkinson's disease (PD). Our optimized conditions for producing homogeneous and mature MOs might provide an advanced patient-specific platform for in vitro disease modeling as well as for drug screening for PD.
Autophagy is an essential cellular mechanism that degrades cytoplasmic proteins and organelles to recycle their components; however, the contribution of autophagy during meiosis has not been studied ...in porcine oocytes maturing in vitro. In this study, we observed that the autophagy-related gene, LC3, was expressed in porcine oocytes during maturation for 44 h in vitro. Knockdown of the autophagy-related gene, BECN1, reduced both BECN1 and LC3 protein expression levels. Moreover, BECN1 knockdown and treatment with the autophagy inhibitor, LY294002, during maturation of porcine oocytes in vitro impaired polar body extrusion, disturbed mitochondrial function, triggered the DNA damage response, and induced early apoptosis in porcine oocytes. Autophagy inhibition during oocyte maturation also impaired the further developmental potential of porcine oocytes. These results indicate that autophagy is required for the in vitro maturation of porcine oocytes.
Nitric oxide (NO) is emerging as a key signalling molecule in plants. The chief mechanism for the transfer of NO bioactivity is thought to be S-nitrosylation, the addition of an NO moiety to a ...protein cysteine thiol to form an S-nitrosothiol (SNO). The enzyme S-nitrosoglutathione reductase (GSNOR) indirectly controls the total levels of cellular S-nitrosylation, by depleting S-nitrosoglutathione (GSNO), the major cellular NO donor. Here we show that depletion of GSNOR function impacts tomato (Solanum lycopersicum. L) fruit development. Thus, reduction of GSNOR expression through RNAi modulated both fruit formation and yield, establishing a novel function for GSNOR. Further, depletion of S. lycopersicum GSNOR (SlGSNOR) additionally impacted a number of other developmental processes, including seed development, which also has not been previously linked with GSNOR activity. In contrast to Arabidopsis, depletion of GSNOR function did not influence root development. Further, reduction of GSNOR transcript abundance compromised plant immunity. Surprisingly, this was in contrast to previous data in Arabidopsis that reported that reducing Arabidopsis thaliana GSNOR (AtGSNOR) expression by antisense technology increased disease resistance. We also show that increased SlGSNOR expression enhanced pathogen protection, uncovering a potential strategy to enhance disease resistance in crop plants. Collectively, our findings reveal, at the genetic level, that some but not all GSNOR activities are conserved outside the Arabidopsis reference system. Thus, manipulating the extent of GSNOR expression may control important agricultural traits in tomato and possibly other crop plants.
Mangiferin (MGN) is primarily found in the fruits, leaves, and bark of plants of the Anacardiaceae family, including mangoes. MGN exhibits various pharmacological effects, such as protection of the ...liver and gallbladder, anti‐lipid peroxidation, and cancer prevention. This study aimed to investigate the effects of MGN supplementation during in vitro culture (IVC) on the antioxidant capacity of early porcine embryos and the underlying mechanisms involved. Porcine parthenotes in the IVC medium were exposed to different concentrations of MGN (0, 0.01, 0.1, and 1 μM). The addition of 0.1 μM MGN significantly increased the blastocyst formation rate of porcine embryos while reducing the apoptotic index and autophagy. Furthermore, the expression of antioxidation‐related (SOD2, GPX1, NRF2, UCHL1), cell pluripotency (SOX2, NANOG), and mitochondria‐related (TFAM, PGC1α) genes was upregulated. In contrast, the expression of apoptosis‐related (CAS3, BAX) and autophagy‐related (LC3B, ATG5) genes decreased after MGN supplementation. These findings suggest that MGN improves early porcine embryonic development by reducing oxidative stress‐related genes.
The body condition of a snake species provides important physiological, morphological, and ecological information that elucidates its habits, life cycle, and competitive relationships. We measured ...the body size and condition of the wild Gloydius ussuriensis population in South Korea from 2018 to 2022, analyzed the degree of intraspecific niche overlap, and identified the geographic and climatic factors affecting their body condition. We found that the females were longer than the males. The body condition index (BCI) of G. ussuriensis differed depending on sex and season; the BCI of the females and males was highest in August and October, respectively. Environmental factors related to altitude and temperature affected the body condition of G. ussuriensis; BCI increased as the mean annual temperature and winter temperature increased; however, it increased when the annual temperature range decreased. The mean Pinaka index was 0.96, indicating a high degree of niche overlap; however, the niche overlap among the neonates was less than that among the adults and juveniles. To elucidate the causes of niche overlap and mechanisms behind the intraspecific competition among G. ussuriensis individuals, the habitat and utilization of food resources at different development stages of G. ussuriensis should be further investigated.
Our previous study established that chrysoeriol (CHE) can reduce reactive oxygen species (ROS) accumulation, apoptosis, and autophagy in vitro culture (IVC) of porcine embryos. However, the role of ...CHE in oocyte maturation and lipid homeostasis is unclear. Herein, we aimed to elucidate the effect of CHE on porcine oocyte competence in vitro maturation (IVM) and subsequent embryo development. The study chooses parthenogenetic activated porcine oocytes as the research model. The study revealed that the cumulus expansion index and related gene expressions are significantly elevated after supplementing 1 μM CHE. Although there were no significant differences in nuclear maturation and cleavage rates, the blastocyst formation rate and total cell numbers were significantly increased in the 1 μM CHE group. In addition, CHE improved the expression of genes related to oocyte and embryo development. ROS was significantly downregulated in all CHE treatment groups, and intracellular GSH (glutathione) was significantly upregulated in 0.01, 0.1, and 1 μM CHE groups. The immunofluorescence results indicated that mitochondrial membrane potential (MMP) and lipid droplet (LD), fatty acid (FA), ATP, and functional mitochondria contents significantly increased with 1 μM CHE compared to the control. Furthermore, CHE increased the expression of genes related to lipid metabolism, mitochondrial biogenesis, and β-oxidation.
Abstract
We investigated the influence of scan direction on subfoveal choroidal vascularity index (CVI) measurements using spectral-domain optical coherence tomography (SD-OCT) in young healthy ...subjects. Seventy-eight eyes of 41 healthy volunteers were included. Choroidal structures were obtained using SD-OCT with enhanced depth imaging (EDI) through radial scans at the center of the macula. The subfoveal choroidal images in the horizontal (0°), 45°, vertical (90°) and − 45° directions were recorded and CVIs were analyzed according to their respective directions using image binarization. Additionally, subfoveal choroidal thickness (SFCT), and axial eye length were measured. The SFCT and subfoveal CVI showed a negative correlation but were only significant for the 45° scan (Pearson’s r = − 0.262, P = 0.021). The axial eye length and subfoveal CVI had no significant correlation in any direction (all P > 0.05). In the Bland–Altman plot, the subfoveal CVI measurement showed high agreement among the four scan directions. When the SFCT was ≥ 300 µm, there was no difference in the measured values of the subfoveal CVI among the four scan directions; however, when the SFCT was < 300 µm, there was a significant difference in subfoveal CVI among the scan directions (one-way analysis of variance, F = 4.685, P = 0.004). In subfoveal CVI measurement, it is considered that the horizontal (0°) scan can represent the vertical (90°) or oblique (45°, − 45°) scans. However, when the SFCT is thinner, the subfoveal CVI in each direction of radial scan may vary significantly. Hence, caution is required in the interpretation.
Brain organoids have been considered as an advanced platform for in vitro disease modeling and drug screening, but numerous roadblocks exist, such as lack of large-scale production technology and ...lengthy protocols with multiple manipulation steps, impeding the industrial translation of brain organoid technology. Here, we describe the high-speed and large-scale production of midbrain organoids using a high-throughput screening-compatible platform within 30 days. Micro midbrain organoids (µMOs) exhibit a highly uniform morphology and gene expression pattern with minimal variability. Notably, µMOs show dramatically accelerated maturation, resulting in the generation of functional µMOs within only 30 days of differentiation. Furthermore, individual µMOs display highly consistent responsiveness to neurotoxin, suggesting their usefulness as an in vitro high-throughput drug toxicity screening platform. Collectively, our data indicate that µMO technology could represent an advanced and robust platform for in vitro disease modeling and drug screening for human neuronal diseases.
Purpose: To investigate the influence of scan direction on subfoveal choroidal vascularity index (CVI) measurements using spectral‐domain optical coherence tomography (SD‐OCT) in young healthy ...subjects.
Methods: Seventy‐eight eyes of 41 healthy volunteers were included. Choroidal structures were obtained using SD‐OCT with enhanced depth imaging (EDI) through radial scans at the center of the macula at 30° × 30°. The subfoveal choroidal images in the horizontal (0°), 45°, vertical (90°) and −45° directions were recorded and CVIs were analysed according to their respective directions, using image binarization. Additionally, subfoveal choroidal thickness (SFCT) and axial eye length were measured.
Results: The SFCT and subfoveal CVI showed a negative correlation but were only significant for the 45° scan (Pearson's r = −0.262, p = 0.021). The axial eye length and subfoveal CVI had no significant correlation in any direction (all p > 0.05). In the Bland–Altman plot, the subfoveal CVI measurement showed high agreement among the four scan directions. When the SFCT was ≥300 μm, there was no difference in the measured values of the subfoveal CVI among the four scan directions; however, when the SFCT was <300 μm, there was a significant difference in subfoveal CVI among the scan directions (one‐way analysis of variance, F = 4.685, p = 0.004).
Conclusions: In subfoveal CVI measurement, it is considered that the horizontal (0°) scan can represent the vertical (90°) or oblique (45°, −45°) scans. However, when the SFCT is thinner, the subfoveal CVI in each direction of radial scan may vary significantly. Hence, caution is required in the interpretation.
Chrysoeriol (CHE) is a flavonoid substance that exists in many plants. It has various physiological and pharmacological effects, including anti-inflammatory, antioxidant, anti-tumor, and protective ...activity, especially for the cardiovascular system and liver. Among common livestock embryos, porcine embryos are often considered high-quality objects for studying the antioxidant mechanisms of oocytes. Because porcine embryos contain high levels of lipids, they are more vulnerable to external stimuli, which affect development. Our study explored the influence of CHE supplementation on oxidative stress in porcine oocytes and its possible mechanisms. Different concentrations of CHE (0, 0.1, 1, and 3 µM) were supplemented in the in vitro culture medium of the porcine oocytes. The results showed that supplementation with 1 µM CHE significantly increased the blastocyst rate and total cell number of embryos in vitro. After finding the beneficial effects of CHE, we measured reactive oxygen species (ROS), glutathione (GSH), and mitochondrial membrane potential (MMP) when the oocytes reached the 4-cell stage of development and determined the levels of apoptosis, cell proliferation, and autophagy at the blastocyst stage of development. The expression levels of some related genes were preliminarily detected by qRT-PCR. The results showed that the apoptosis of blastocysts in the CHE-treated culture also decreased compared with the untreated culture. Furthermore, CHE downregulated intracellular ROS and increased GSH in the embryos. CHE was also shown to improve the activity of mitochondria and inhibit the occurrence of autophagy. In addition, antioxidant-related genes (SOD1, SOD2, and CAT) and cell pluripotency-related genes (SOX2, OCT4, and NANOG) were upregulated. At the same time, apoptosis-related (Caspase 3) and autophagy-related (LC3B) genes showed a downward trend after supplementation with CHE. These results indicate that CHE improved the development of porcine embryos in vitro by reducing oxidative stress and autophagy levels.
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