Integrinâ Targeted Cyclic Forces Accelerate Neural Tubeâ Like Rosette Formation from Human Embryonic Stem Cells

Mechanical forces play important roles in human embryonic stem cell (hESC) differentiation. To investigate the impact of dynamic mechanical forces on neural induction of hESCs, this study employs acoustic tweezing cytometry (ATC) to apply cyclic forces/strains to hESCs by actuating integrinâ bound microbubbles using ultrasound pulses. Accelerated neural induction of hESCs is demonstrated as the result of combined action of ATC and neural induction medium (NIM). Specifically, application of ATC for 30 min followed by culture in NIM upregulates neuroecdoderm markers Pax6 and Sox1 as early as 6 h after ATC, and induces neural tubeâ like rosette formation at 48 h after ATC. In contrast, no changes are observed in hESCs cultured in NIM without ATC treatment. In the absence of NIM, ATC application decreases Oct4, but does not increase Pax6 and Sox1 expression, nor does it induce neural rossette formation. The effects of ATC are abolished by inhibition of FAK, myosin activity, and RhoA/ROCK signaling. Taken together, the results reveal a synergistic action of ATC and NIM as an integrated mechanobiology mechanism that requires both integrinâ targeted cyclic forces and chemical factors for accelerated neural induction of hESCs.Acoustic tweezing cytometry (ATC) applies integrinâ targeted cyclic forces/strains to human embryonic stem cells (hESCs). Dependent on FAK, myosin activity, and RhoA/ROCK signaling, synergistic action of ATC for 30 min and neural induction medium greatly accelerates neural induction of hESCs, resulting in upregulated neuroecdoderm markers Pax6 and Sox1 by 6 h and neural tubeâ like rosette formation at 48 h.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/151881/1/adbi201900064_am.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/151881/2/adbi201900064.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/151881/3/adbi201900064-sup-0001-SuppMat.pd

Acoustic Tweezing Cytometry Induces Rapid Initiation of Human Embryonic Stem Cell Differentiation.

Mechanical forces play critical roles in influencing human embryonic stem cell (hESC) fate. However, it remains largely uncharacterized how local mechanical forces influence hESC behavior in vitro. Here, we used an ultrasound (US) technique, acoustic tweezing cytometry (ATC), to apply targeted cyclic subcellular forces to hESCs via integrin-bound microbubbles (MBs). We found that ATC-mediated cyclic forces applied for 30 min to hESCs near the edge of a colony induced immediate global responses throughout the colony, suggesting the importance of cell-cell connection in the mechanoresponsiveness of hESCs to ATC-applied forces. ATC application generated increased contractile force, enhanced calcium activity, as well as decreased expression of pluripotency transcription factors Oct4 and Nanog, leading to rapid initiation of hESC differentiation and characteristic epithelial-mesenchymal transition (EMT) events that depend on focal adhesion kinase (FAK) activation and cytoskeleton (CSK) tension. These results reveal a unique, rapid mechanoresponsiveness and community behavior of hESCs to integrin-targeted cyclic forces

Belowground Rhizomes in Paleosols: The Hidden Half of an Early Devonian Vascular Plant

The colonization of terrestrial environments by rooted vascular plants had far-reaching impacts on the Earth system. However, the belowground structures of early vascular plants are rarely documented, and thus the plant−soil interactions in early terrestrial ecosystems are poorly understood. Here we report the earliest rooted paleosols (fossil soils) in Asia from Early Devonian deposits of Yunnan, China. Plant traces are extensive within the soil and occur as complex network-like structures, which are interpreted as representing long-lived, belowground rhizomes of the basal lycopsid Drepanophycus. The rhizomes produced large clones and helped the plant survive frequent sediment burial in well-drained soils within a seasonal wet−dry climate zone. Rhizome networks contributed to the accumulation and pedogenesis of floodplain sediments and increased the soil stabilizing effects of early plants. Predating the appearance of trees with deep roots in the Middle Devonian, plant rhizomes have long functioned in the belowground soil ecosystem. This study presents strong, direct evidence for plant−soil interactions at an early stage of vascular plant radiation. Soil stabilization by complex rhizome systems was apparently widespread, and contributed to landscape modification at an earlier time than had been appreciated

Acoustic Actuation of Integrin‐Bound Microbubbles for Mechanical Phenotyping during Differentiation and Morphogenesis of Human Embryonic Stem Cells

Early human embryogenesis is a dynamic developmental process, involving continuous and concomitant changes in gene expression, structural reorganization, and cellular mechanics. However, the lack of investigation methods has limited the understanding of how cellular mechanical properties change during early human embryogenesis. In this study, ultrasound actuation of functionalized microbubbles targeted to integrin (acoustic tweezing cytometry, ATC) is employed for in situ measurement of cell stiffness during human embryonic stem cell (hESC) differentiation and morphogenesis. Cell stiffness, which is regulated by cytoskeleton structure, remains unchanged in undifferentiated hESCs, but significantly increases during neural differentiation. Further, using the recently established in vitro 3D embryogenesis models, ATC measurements reveal that cells continue to stiffen while maintaining pluripotency during epiblast cyst formation. In contrast, during amniotic cyst formation, cells first become stiffer during luminal cavity formation, but softens significantly when cells differentiate to form amniotic cysts. These results suggest that cell stiffness changes not only due to 3D spatial organization, but also with cell fate change. ATC therefore provides a versatile platform for in situ measurement of cellular mechanical property, and cell stiffness may be used as a mechanical biomarker for cell lineage diversification and cell fate specification during embryogenesis.Ultrasound actuation of functionalized microbubbles targeted to integrin (acoustic tweezing cytometry) is employed for in situ measurement of cell stiffness during human embryonic stem cell neural differentiation and morphogenesis in 3D embryogenesis model. The results suggest that cell stiffness changes not only due to 3D spatial organization, but also with cell fate change.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/146940/1/smll201803137.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/146940/2/smll201803137_am.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/146940/3/smll201803137-sup-0001-S1.pd

Reward prediction error in learning-related behaviors

Learning is a complex process, during which our opinions and decisions are easily changed due to unexpected information. But the neural mechanism underlying revision and correction during the learning process remains unclear. For decades, prediction error has been regarded as the core of changes to perception in learning, even driving the learning progress. In this article, we reviewed the concept of reward prediction error, and the encoding mechanism of dopaminergic neurons and the related neural circuities. We also discussed the relationship between reward prediction error and learning-related behaviors, including reversal learning. We then demonstrated the evidence of reward prediction error signals in several neurological diseases, including Parkinson’s disease and addiction. These observations may help to better understand the regulatory mechanism of reward prediction error in learning-related behaviors

Transcription-associated mutation promotes RNA complexity in highly expressed genes - a major new source of selectable variation

Alternatively spliced transcript isoforms are thought to play a critical role for functional diversity. However, the mechanism generating the enormous diversity of spliced transcript isoforms remains unknown, and its biological significance remains unclear. We analyzed transcriptomes in saker falcons, chickens, and mice to show that alternative splicing occurs more frequently, yielding more isoforms, in highly expressed genes. We focused on hemoglobin in the falcon, the most abundantly expressed genes in blood, finding that alternative splicing produces 10-fold more isoforms than expected from the number of splice junctions in the genome. These isoforms were produced mainly by alternative use of de novo splice sites generated by transcription-associated mutation (TAM), not by the RNA editing mechanism normally invoked. We found that high expression of globin genes increases mutation frequencies during transcription, especially on nontranscribed DNA strands. After DNA replication, transcribed strands inherit these somatic mutations, creating de novo splice sites, and generating multiple distinct isoforms in the cell clone. Bisulfate sequencing revealed that DNA methylation may counteract this process by suppressing TAM, suggesting DNA methylation can spatially regulate RNA complexity. RNA profiling showed that falcons living on the high Qinghai–Tibetan Plateau possess greater global gene expression levels and higher diversity of mean to high abundance isoforms (reads per kilobases per million mapped reads ≥18) than their low-altitude counterparts, and we speculate that this may enhance their oxygen transport capacity under low-oxygen environments. Thus, TAM-induced RNA diversity may be physiologically significant, providing an alternative strategy in lifestyle evolution

Belowground rhizomes in paleosols:The hidden half of an Early Devonian vascular plant

The colonization of terrestrial environments by rooted vascular plants had far-reaching impacts on the Earth system. However, the belowground structures of early vascular plants are rarely documented, and thus the plant-soil interactions in early terrestrial ecosystems are poorly understood. Here we report the earliest rooted paleosols (fossil soils) in Asia from Early Devonian deposits of Yunnan, China. Plant traces are extensive within the soil and occur as complex network-like structures, which are interpreted as representing long-lived, belowground rhizomes of the basal lycopsid Drepanophycus. The rhizomes produced large clones and helped the plant survive frequent sediment burial in well-drained soils within a seasonal wet-dry climate zone. Rhizome networks contributed to the accumulation and pedogenesis of floodplain sediments and increased the soil stabilizing effects of early plants. Predating the appearance of trees with deep roots in the Middle Devonian, plant rhizomes have long functioned in the belowground soil ecosystem. This study presents strong, direct evidence for plant-soil interactions at an early stage of vascular plant radiation. Soil stabilization by complex rhizome systems was apparently widespread, and contributed to landscape modification at an earlier time than had been appreciated.National Natural Science Foundation of China [41272018]; Yunnan Key Laboratory for Palaeobiology, Yunnan University [2015DG007-KF04]; Key Laboratory of Economic Stratigraphy and Palaeogeography, Chinese Academy of Sciences (Nanjing Institute of Geology and Palaeontology)SCI(E)[email protected]

Experimental and analytical investigation on flexural behaviour of RC beams strengthened with NSM CFRP prestressed concrete prisms

This investigation aims to study the flexural behaviour of reinforced concrete (RC) beams strengthened with near-surface mounted (NSM) carbon fibre reinforced polymer prestressed concrete prisms (CFRP-PCPs). Eight RC beams were tested under monotonic loading until the failure load was reached. One beam was un-strengthened to act as a control beam. The other seven beams were strengthened with non-prestressed or prestressed NSM CFRP-PCPs. The effects of bond length, prestress level, and concrete type of the CFRP-PCPs on the flexural capacity, flexural crack and deflection are discussed in this paper. The results indicate that the flexural capacity of RC beams strengthened with NSM CFRP-PCPs was greater than the control beam. An obvious improvement was discovered in the crack resistance when the RC beams were strengthened with prestressed NSM CFRP-PCPs. The strengthened beams showed a higher first-cracking, yielding, and ultimate load as the bond length and prestress level of CFRP-PCPs increased up to a critical level. The beams strengthened with CFRP-PCPs, which were cast with ultra-high performance concrete (UHPC), exhibited greater load capacity than the corresponding beams with epoxy resin mortar. The analytical model of flexural response for the NSM CFRP-PCPs strengthening beams is presented. The analytical results are in good agreement with the experimental results, which revealed the NSM CFRP-PCPs is an effective technique for flexural strengthening of the RC beams