Dataset Information

Oocyte Penetration Speed Optimization Based on Intracellular Strain.

ABSTRACT: Oocyte penetration is an essential step for many biological technologies, such as animal cloning, embryo microinjection, and intracytoplasmic sperm injection (ICSI). Although the success rate of robotic cell penetration is very high now, the development potential of oocytes after penetration has not been significantly improved compared with manual operation. In this paper, we optimized the oocyte penetration speed based on the intracellular strain. We firstly analyzed the intracellular strain at different penetration speeds and performed the penetration experiments on porcine oocytes. Secondly, we studied the cell development potential after penetration at different penetration speeds. The statistical results showed that the percentage of large intracellular strain decreased by 80% and the maximum and average intracellular strain decreased by 25-38% at the penetration speed of 50 μm/s compared to at 10 μm/s. Experiment results showed that the cleavage rates of the oocytes after penetration increased from 65.56% to 86.36%, as the penetration speed increased from 10 to 50 μm/s. Finally, we verified the gene expression of oocytes after penetration at different speeds. The experimental results showed that the totipotency and antiapoptotic genes of oocytes were significantly higher after penetration at the speed of 50 μm/s, which verified the effectiveness of the optimization method at the gene level.

SUBMITTER: Liu Y

PROVIDER: S-EPMC8875814 | biostudies-literature | 2022 Feb

REPOSITORIES: biostudies-literature

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Oocyte Penetration Speed Optimization Based on Intracellular Strain.

Liu Yaowei Y Cui Maosheng M Zhang Yidi Y Zhao Xiangfei X Sun Mingzhu M Zhao Xin X

Micromachines 20220217 2

Oocyte penetration is an essential step for many biological technologies, such as animal cloning, embryo microinjection, and intracytoplasmic sperm injection (ICSI). Although the success rate of robotic cell penetration is very high now, the development potential of oocytes after penetration has not been significantly improved compared with manual operation. In this paper, we optimized the oocyte penetration speed based on the intracellular strain. We firstly analyzed the intracellular strain at ...[more]

PMID: 35208433

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Project description:Intestinal mucus is the first line of defense against intestinal pathogens. It acts as a physical barrier between epithelial tissues and the lumen that enteropathogens must overcome to establish a successful infection. We investigated the motile behavior of two Vibrio cholerae strains (El Tor C6706 and Classical O395) in mucus using single-cell tracking in unprocessed porcine intestinal mucus. We determined that V. cholerae can penetrate mucus using flagellar motility and that alkaline pH increases swimming speed and, consequently, improves mucus penetration. Microrheological measurements indicate that changes in pH between 6 and 8 (the physiological range for the human small intestine) had little effect on the viscoelastic properties of mucus. Finally, we determined that acidic pH promotes surface attachment by activating the mannose-sensitive hemagglutinin (MshA) pilus in V. cholerae El Tor C6706 without a measurable change in the total cellular concentration of the secondary messenger cyclic dimeric GMP (c-di-GMP). Overall, our results support the hypothesis that pH is an important factor affecting the motile behavior of V. cholerae and its ability to penetrate mucus. Therefore, changes in pH along the human small intestine may play a role in determining the preferred site for V. cholerae during infection.IMPORTANCE The diarrheal disease cholera is still a burden for populations in developing countries with poor sanitation. To develop effective vaccines and prevention strategies against Vibrio cholerae, we must understand the initial steps of infection leading to the colonization of the small intestine. To infect the host and deliver the cholera toxin, V. cholerae has to penetrate the mucus layer protecting the intestinal tissues. However, the interaction of V. cholerae with intestinal mucus has not been extensively investigated. In this report, we demonstrated using single-cell tracking that V. cholerae can penetrate intestinal mucus using flagellar motility. In addition, we observed that alkaline pH improves the ability of V. cholerae to penetrate mucus. This finding has important implications for understanding the dynamics of infection, because pH varies significantly along the small intestine, between individuals, and between species. Blocking mucus penetration by interfering with flagellar motility in V. cholerae, reinforcing the mucosa, controlling intestinal pH, or manipulating the intestinal microbiome will offer new strategies to fight cholera.

Dataset Information

Oocyte Penetration Speed Optimization Based on Intracellular Strain.

Publications

Oocyte Penetration Speed Optimization Based on Intracellular Strain.

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