A systematic review and meta-analysis on the root effects and toxic mechanisms of rare earth elements.

Rare earth elements (REEs) have attracted much attention because of their unique physical and chemical properties. The root system is the plant organ most directly in contact with REEs, and it is critical to understand the mechanisms of interaction between the two. This paper investigates the effects of REEs on plant enrichment and fractionation, as well as on various developmental and toxicity indices of the root system. REEs are more likely to be deposited on the root surface under the influence of root secretion. The complexation between the two affects the uptake and fractionation of REEs and the altered pattern of root secretion. The toxicity mechanisms of REEs on plant root cells were lied in: (1) REEs generate reactive oxygen species after entering the plant, leading to oxidative stress and damage to plant cells; (2) REEs with higher charge-to-volume ratios compete for organic ligands with or displace Ca2+, further disrupting the normal function of plant root cells. It was shown that the sensitivity of inter-root microorganisms to REEs varied depending on the content and physicochemical properties of REEs. The paper also concluded with a meta-analysis of phytotoxicity induced by REEs, which showed that REEs affect plant physiological parameters. REEs, as a source of oxidative stress, triggered lipid peroxidation damage in plants and enhanced the activity of antioxidant enzymes, thus revealing the significant toxicity of REEs to plants. The phytotoxic effects of REEs increased with time and concentration. These results help to elucidate the ecotoxicology of rare earth-induced phytotoxicity.

Cortical somatostatin long-range projection neurons and interneurons exhibit divergent developmental trajectories.

The mammalian cerebral cortex contains an extraordinary diversity of cell types that emerge by implementing different developmental programs. Delineating when and how cellular diversification occurs is particularly challenging for cortical inhibitory neurons because they represent a small proportion of all cortical cells and have a protracted development. Here, we combine single-cell RNA sequencing and spatial transcriptomics to characterize the emergence of neuronal diversity among somatostatin-expressing (SST+) cells in mice. We found that SST+ inhibitory neurons segregate during embryonic stages into long-range projection (LRP) neurons and two types of interneurons, Martinotti cells and non-Martinotti cells, following distinct developmental trajectories. Two main subtypes of LRP neurons and several subtypes of interneurons are readily distinguishable in the embryo, although interneuron diversity is likely refined during early postnatal life. Our results suggest that the timing for cellular diversification is unique for different subtypes of SST+ neurons and particularly divergent for LRP neurons and interneurons.

Silencing of episomal transgene expression in liver by plasmid bacterial backbone DNA is independent of CpG methylation.

Minicircle DNA vectors devoid of plasmid bacterial backbone, (BB) DNAs, are transcriptionally persistent, whereas their parent plasmid counterparts are silenced in the liver. In this study we establish that circular plasmid BB provided in trans did not silence a transgene expression cassette in vivo, further confirming our previous conclusions that the covalent attachment of the plasmid BB to the expression cassette is required for DNA silencing. Given the high concentration of CpG dinucleotides in the plasmid BB, we investigated the role of DNA methylation on transgene silencing in vivo. The presence or absence of methylation in CpG motifs in routine plasmid BBs had no significant effect on transcriptional silencing. Furthermore, the removal of the CpG motifs from the BB did not ameliorate transcriptional silencing. Transgene silencing was partially inhibited when two tandem copies of the chicken cHS4 insulator at each end of a routine plasmid vector were used. These results are consistent with the idea that the transcriptional repression observed with plasmid DNA vectors in the liver is caused by formation of repressive heterochromatin on the plasmid DNA backbone, which then spreads and inactivates the transgene in cis, and that CpG content or methylation has little or no influence in the process.

Histone modifications are associated with the persistence or silencing of vector-mediated transgene expression in vivo.

One of the major obstacles to success in non-viral gene therapy is transcriptional silencing of the DNA vector. The mechanisms underlying gene silencing/repression in mammalian cells are complex and remain unclear. Because changes in chromatin structure and, in particular, histone modifications are involved in transcriptional regulation of endogenous genes, we hypothesized that changes in the pattern of histone modifications were related to the observed transcriptional silencing of exogenous DNA vectors. We used antibodies against specific modified histones to perform chromatin immunoprecipitation (ChIP) analyses on liver lysates from mice transfected with two types of plasmids: (i) DNA minicircles (MCs) devoid of bacterial plasmid backbone DNA, which showed marked persistence of transgene expression, and (ii) their parental plasmids, which were silenced over time. Silencing of the transgene from the parental vectors was accompanied by an increase in heterochromatin-associated histone modifications and a decrease in modifications typically associated with euchromatin. Conversely, the pattern of histone modifications on the MC DNA was consistent with euchromatin. Our data indicates that (i) episomal vectors undergo chromatinization in vivo, and (ii) both persistence and silencing of transgene expression are associated with specific histone modifications.