Protocol for intra-nerve AAV injection and dorsal root potential recording for optogenetic modulation of the peripheral sensory nerve activity [0.03%]
Akito Kosugi,Wupuer Sidikejiang,Shinji Kubota et al.
Akito Kosugi et al.
Optogenetic modulation of peripheral sensory nerve activity holds great potential for revealing the mechanisms underlying sensory disorders, with important implications for developing therapeutic strategies. Here, we present a protocol for ...
Satoshi Morita,Takayuki Kondo,Haruhisa Inoue
Satoshi Morita
Here, we present a protocol for describing the differentiation of human cortical neurons from induced pluripotent stem cells (iPSCs) and the subsequent analysis of lipid membrane fluidity using the LipiORDER fluorescent probe. We describe s...
Protocol for genetic manipulation of cervical lymph node-innervating nociceptive neurons in mice via retrograde viral tracing [0.03%]
Yibo Guo,Tong Ji,Yu Zhang
Yibo Guo
This protocol details the surgical procedure for genetically manipulating cervical lymph node (LN)-innervating nociceptive neurons in mice. We describe steps for exposing cervical LNs, microinjecting retrograde adeno-associated viruses (AAV...
Protocol for preparing tyramine functionalized hyaluronic acid via aqueous DMTMM-mediated conjugation [0.03%]
Gregor Miklosic,Daphne van der Heide,Claudia Loebel et al.
Gregor Miklosic et al.
Hyaluronic acid is a ubiquitous glycosaminoglycan with numerous biological and structural functions in the body. Here, we present a protocol for the practical and efficient functionalization of HA with tyramine. We describe the reaction con...
Protocol to measure transcriptional bursting of endogenous genes using high-throughput RNA-FISH [0.03%]
Varun Sood,Tom Misteli
Varun Sood
We present a protocol to measure transcriptional bursting of endogenous human genes that is widely applicable to adherent cells. We describe the steps for nascent RNA fluorescence in situ hybridization (nscRNA-FISH) in a high-throughput ima...
Protocol to assess calcium signaling in response to metabolic alterations by flow cytometry and live-cell imaging [0.03%]
Laura Schlautmann,Sven Burgdorf
Laura Schlautmann
Calcium (Ca2+) signaling is essential to cellular processes, such as signal transduction and migration, and dysregulation of Ca2+ homeostasis can impact cellular function. Here, we present a protocol to analyze Ca2+ signaling by its release...
Zhihuang Liang,Shufang Chen,Tiezhu Shi et al.
Zhihuang Liang et al.
Renal cancer is characterized by an insidious onset and a high tendency for lung metastasis in advanced stages. Here, we present a protocol for constructing an orthotopic mouse model that recapitulates the spontaneous metastasis process of ...
Protocol for visualizing microglial lysosomal content by immunohistochemistry in the rodent brain [0.03%]
Celeste Laporte,Masha Prager-Khoutorsky
Celeste Laporte
Microglia, the resident phagocytes of the central nervous system, clear diverse substrates in development, aging, injury, and disease. Here, we present a protocol to visualize phagocytosed content within microglial lysosomes in the rodent b...
Protocol for targeted peripheral adeno-associated viral vector transduction of the intradental neurons in mice [0.03%]
Ling-Yu Liu,Amy Chang,Isaac T Berthaume et al.
Ling-Yu Liu et al.
Here, we present the methodology for surgical preparation of the mouse molar teeth and subsequent adeno-associated virus (AAV) transduction of intradental neurons within the tooth (AAV-i.d.). We describe steps for mouse positioning, microsc...
Lucy Funes,Frederick R Maxfield,Santiago Solé-Domènech
Lucy Funes
Microglia use digestive exophagy to partially degrade, extracellularly, Alzheimer's amyloid-beta aggregates that are too large to be phagocytosed. Here, we present a protocol to quantify this mechanism in cell culture. We describe steps for...