Dissociated cell cultures of the rodent hippocampus have become a standard model for studying many facets of neural development, including the development of polarity, axonal and dendritic growth, and synapse formation. The cultures are quite homogeneous--∼90% of the cells are pyramidal neurons--and it is relatively easy to express green fluorescent protein (GFP)-tagged proteins by transfection. This article describes the cultures and the key features of the system used to image them. It also includes suggestions on labeling cells with GFP-tagged proteins. It concludes with a discussion of the advantages and disadvantages of this culture system.
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http://dx.doi.org/10.1101/pdb.ip068221 | DOI Listing |
Arch Microbiol
January 2025
Agricultural Botany Department, Faculty of Agriculture, Suez Canal University, 41522, Ismailia, Egypt.
Researchers have reported that Bacillus megaterium BM18-2 reduces Cd toxicity in Hybrid Pennisetum, but understanding the interaction between plants and associated endophytes is crucial for understanding phytoremediation strategies under heavy metal stress. The current study aims to monitor the colonization patterns of GFP-labeled endophytic bacteria BM18-2 on Hybrid Pennisetum grass. Additionally, it will monitor Cd's effect on plant bacterial colonization.
View Article and Find Full Text PDFGenes (Basel)
December 2024
Faculty of Frontiers of Innovative Research in Science and Technology (FIRST), Konan University, Kobe 650-0047, Japan.
Background/objectives: Actin plays a crucial role not only in the cytoplasm, but also in the nucleus, influencing various cellular behaviors, including cell migration and gene expression. Recent studies reveal that nuclear actin dynamics is altered by cellular stresses, such as DNA damage; however, the effect of heat shock on nuclear actin dynamics, particularly in the nucleolus, remains unclear. This study aims to elucidate the contribution of nucleolar actin to cellular responses under heat shock conditions.
View Article and Find Full Text PDFBiomolecules
December 2024
Department of Neurosurgery, Zhongnan Hospital of Wuhan University, School of Pharmaceutical Sciences, Wuhan University, Wuhan 430071, China.
Hyaluronan (HA) is one of the crucial components of the extracellular matrix in vertebrates and is synthesized by three hyaluronan synthases (HASs), namely HAS1, HAS2, and HAS3. The low expression level of HASs in normal keratinocytes and other various types of cells presents a recognized challenge, impeding biological and pathological research on their localization. In this study, the human proteins HAS1, HAS2, and HAS3 with fused maltose-binding protein (MBP) tags were successfully expressed at high levels and purified for the first time in HEK293F cells.
View Article and Find Full Text PDFProc Natl Acad Sci U S A
December 2024
Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, NY 10029.
mBio
December 2024
Tumour Virology, International Centre for Genetic Engineering and Biotechnology, Trieste, Italy.
Unlabelled: Previous studies have shown that E6 interacts with the E6-associated protein (E6AP) ubiquitin-protein ligase and directs its ubiquitylation activity toward several specific cellular proteins, one of the most important of which is p53. Interestingly, E6AP not only aids in the E6-directed degradation of cellular substrates but also stabilizes the E6 protein by protecting it from proteasome-mediated degradation. However, there is no information available about the ubiquitin ligases that regulate the stability and activity of the human papillomavirus (HPV) E6 oncoprotein in the absence of E6AP.
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