Astrocytes are glial cells of the central nervous system that modulate neuronal function. Here, we present glyoxal-fixed astrocyte nuclei transcriptomics (GFAT), a protocol for the purification and transcriptomic analysis of astrocyte nuclei from the cortex and cerebellum of adult and aged fresh mouse brain. We describe steps for tissue dissection, glyoxal fixation, homogenization, nuclei isolation, antibody staining, fluorescence-activated cell sorting, and RT-qPCR or bulk RNA sequencing. GFAT does not require transgenic lines or viral injection and allows parallel astrocyte and neuron profiling.
Download full-text PDF |
Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10522990 | PMC |
| http://dx.doi.org/10.1016/j.xpro.2023.102599 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Protocol for detecting eDNA in ecological rare fish using RPA-CRISPR-Cas12a technology.
STAR Protoc
January 2025
School of Public Health, Chongqing Medical University, Chongqing 400016, China; Chongqing Miankai Biotechnology Research Institute Co., Ltd., Chongqing 400025, China. Electronic address:
The recombinase polymerase amplification (RPA)-CRISPR-Cas12a-FQ system enables sensitive detection of environmental DNA (eDNA) in rare fish species. Here, we present a protocol for eDNA amplification and Cas12a for target recognition using RPA. We describe steps for identifying a target site, synthesis and purification of CRISPR RNA (crRNA), and RPA isothermal amplification.
View Article and Find Full Text PDFProtocol for the purification of the plastid-encoded RNA polymerase from transplastomic tobacco plants.
STAR Protoc
January 2025
National Key Laboratory of Crop Genetic Improvement and National Centre of Plant Gene Research, Huazhong Agricultural University, Wuhan 430070, China. Electronic address:
The plastid-encoded RNA polymerase (PEP) plays an essential role in the transcription of the chloroplast genome. Here, we present a strategy to purify the transcriptionally active protein complex from transplastomic tobacco (Nicotiana tabacum) lines in which one of the PEP core subunits is fused to an epitope tag. We describe experimental procedures for designing transformation constructs for PEP purification, selection, and analysis of transplastomic tobacco plants.
View Article and Find Full Text PDFProtocol for high-yield bacterial expression and purification of the voltage-dependent anion channel 1 for high-throughput biophysical assays.
STAR Protoc
January 2025
Department of Molecular Medicine, University of Pavia, Pavia, Italy. Electronic address:
Voltage-dependent anion channel 1 (VDAC1) is a key protein in cellular metabolism and apoptosis. Here, we present a protocol to express and purify milligram amounts of recombinant VDAC1 in Escherichia coli. We detail steps for a fluorescence polarization-based high-throughput screening assay using NADH displacement, along with procedures for thermostability, fluorescence polarization, and X-ray crystallography.
View Article and Find Full Text PDFHeterologous Expression and Optimization of Fermentation Conditions for Recombinant Ikarugamycin Production.
Biotechnol Bioeng
January 2025
Chair of Technical Biochemistry, Technische Universität Dresden, Dresden, Saxony, Germany.
Ikarugamycin is a member of the natural product family of the polycyclic tetramate macrolactams (PoTeMs). The compound exhibits a diverse range of biological activities, including antimicrobial, antiprotozoal, anti-leukemic, and anti-inflammatory properties. In addition, it interferes with several crucial cellular functions, such as oxidized low-density lipoprotein uptake in macrophages, Nef-induced CD4 cell surface downregulation, and mechanisms of endocytosis.
View Article and Find Full Text PDFChromogenic bacterial staining of teeth: a scoping review.
BMC Oral Health
January 2025
Basic Medical and Dental Sciences Department, College of Dentistry, Ajman University, Ajman, UAE.
Article Synopsis
- This scoping review aims to investigate the causes, clinical traits, and treatment methods for chromogenic staining of teeth, following the PRISMA 2022 guidelines.
- A systematic search across multiple medical databases led to the identification of 989 articles, with 21 ultimately included for analysis, highlighting that chromogenic bacteria can cause staining in 3.1 to 18.5% of cases, primarily resulting in black discoloration.
- The findings indicate a lack of comprehensive literature on this topic, emphasizing the need for further research into effective management strategies for these challenging stains linked to specific oral bacteria.
Want AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!