Loss of SHROOM3 affects neuroepithelial cell shape through regulating cytoskeleton proteins in cynomolgus monkey organoids.

Neural tube defects (NTDs) are severe congenital neurodevelopmental disorders arising from incomplete neural tube closure. Although folate supplementation has been shown to mitigate the incidence of NTDs, some cases, often attributable to genetic factors, remain unpreventable. The SHROOM3 gene has been implicated in NTD cases that are unresponsive to folate supplementation; at present, however, the underlying mechanism remains unclear. Neural tube morphogenesis is a complex process involving the folding of the planar epithelium of the neural plate. To determine the role of SHROOM3 in early developmental morphogenesis, we established a neuroepithelial organoid culture system derived from cynomolgus monkeys to closely mimic the in vivo neural plate phase. Loss of SHROOM3 resulted in shorter neuroepithelial cells and smaller nuclei. These morphological changes were attributed to the insufficient recruitment of cytoskeletal proteins, namely fibrous actin (F-actin), myosin II, and phospho-myosin light chain (PMLC), to the apical side of the neuroepithelial cells. Notably, these defects were not rescued by folate supplementation. RNA sequencing revealed that differentially expressed genes were enriched in biological processes associated with cellular and organ morphogenesis. In summary, we established an authentic in vitro system to study NTDs and identified a novel mechanism for NTDs that are unresponsive to folate supplementation.

Functional Connectivity-Based Parcellation of the Thalamus: An Unsupervised Clustering Method and Its Validity Investigation.

Node definition or delineating how the brain is parcellated into individual functionally related regions is the first step to accurately map the human connectome. As a result, parcellation of the human brain has drawn considerable attention in the field of neuroscience. The thalamus is known as a relay in the human brain, with its nuclei sending fibers to the cortical and subcortical regions. Functional magnetic resonance imaging techniques offer a way to parcellate the thalamus in vivo based on its connectivity properties. However, the parcellations from previous studies show that both the number and the distribution of thalamic subdivisions vary with different cortical segmentation methods. In this study, we used an unsupervised clustering method that does not rely on a priori information of the cortical segmentation to parcellate the thalamus. Instead, this approach is based on the intrinsic resting-state functional connectivity profiles of the thalamus with the whole brain. A series of cluster solutions were obtained, and an optimal solution was determined. Furthermore, the validity of our parcellation was investigated through the following: (1) identifying specific resting-state connectivity patterns of thalamic parcels with different brain networks and (2) investigating the task activation and psychophysiological interactions of specific thalamic clusters during 8-Hz flashing checkerboard stimulation with simultaneous finger tapping. Together, the current study provides a reliable parcellation of the thalamus and enhances our understating of thalamic. Furthermore, the current study provides a framework for parcellation that could be potentially extended to other subcortical and cortical regions.

Investigation of the dermal sensitizing potential of traditional medical extracts in local lymph node assays.

Traditional medical extracts are commonly used as complex mixtures, which may contain naturally occurring contact sensitizers. In this investigation, the mice local lymph node assay (LLNA) was performed to evaluate the dermal sensitization potential of Myrrh, Borneolum, Olibanum, Moschus and Cassia Bark, which are widely used in topical traditional medication. In the radioactive LLNA, the stimulation index (SI) values were calculated for each medical extract. Myrrh, Borneolum, Olibanum and Moschus induced dose-dependent cell proliferation and SI was more than 3. Cassia Bark showed no positive response over the range of test concentrations. In the flow cytometry analysis, the total number of CD3(+), CD4(+), and CD8(+) cells in local lymph nodes was increased in Moschus-, Olibanum-, Myrrh- and Borneolum-treated mice. The ratio of the B220(+)/CD3(+) (B/T cell ratio) and the percentage of I-A(k+) cells that was also positive for the CD69 marker (I-A(k+)/ CD69(+)) were increased in the Moschus-, Olibanum- and Myrrh-treated mice. However, no ofbvious change was observed in Borneolum-treated mice. Cassia Bark did not induce changes in the lymphocyte subpopulations. These results indicate that Moschus, Olibanum and Myrrh can be regarded as sensitizers, and Borneolum regarded as an irritant. Cassia Bark is neither a sensitizer nor an irritant. The combination of radioactive and flow cytometric LLNA can be used for the prediction of sensitizing potential of medical extracts which lead to allergic contact dermatitis in humans.