Protocol to record and quantify the intracellular pH in contracting cardiomyocytes

Lyu, Yankun; Timofeyev, Valeriy; Overton, James; Thai, Phung N.; Yamoah, Ebenezer N.; Chiamvimonvat, Nipavan; Zhang, Xiao-Dong

doi:10.1016/j.xpro.2022.101301

Cited by 2 publications

(2 citation statements)

References 18 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It has been reported that CREB signaling activation promotes glucose metabolism in acidified pH i conditions [ 21 ]. Therefore, to study the pH i changes in WT or Nhe1 cKO microglia, we conducted flow cytometry with a novel pH indicator pHrodo whose fluorescent intensity is inversely correlated with pH i [ 22 ] (Fig. 4 a).…”

Section: Resultsmentioning

confidence: 99%

Deletion of Slc9a1 in Cx3cr1+ cells stimulated microglial subcluster CREB1 signaling and microglia-oligodendrocyte crosstalk

Song,

Oft,

Metwally

et al. 2024

J Neuroinflammation

View full text Add to dashboard Cite

Microglial Na/H exchanger-1 (NHE1) protein, encoded by Slc9a1, plays a role in white matter demyelination of ischemic stroke brains. To explore underlying mechanisms, we conducted single cell RNA-seq transcriptome analysis in conditional Slc9a1 knockout (cKO) and wild-type (WT) mouse white matter tissues at 3 days post-stroke. Compared to WT, Nhe1 cKO brains expanded a microglial subgroup with elevated transcription of white matter myelination genes including Spp1, Lgals3, Gpnmb, and Fabp5. This subgroup also exhibited more acidic pHi and significantly upregulated CREB signaling detected by ingenuity pathway analysis and flow cytometry. Moreover, the Nhe1 cKO white matter tissues showed enrichment of a corresponding oligodendrocyte subgroup, with pro-phagocytosis and lactate shuffling gene expression, where activated CREB signaling is a likely upstream regulator. These findings demonstrate that attenuation of NHE1-mediated H+ extrusion acidifies microglia/macrophage and may underlie the stimulation of CREB1 signaling, giving rise to restorative microglia-oligodendrocyte interactions for remyelination.

show abstract

Section: Resultsmentioning

confidence: 99%

Deletion of Slc9a1 in Cx3cr1+ cells stimulated microglial subcluster CREB1 signaling and microglia-oligodendrocyte crosstalk

Song,

Oft,

Metwally

et al. 2024

J Neuroinflammation

View full text Add to dashboard Cite

show abstract

“…It has been reported that CREB signaling activation promotes glucose metabolism in acidi ed pH i conditions (21). Therefore, to study the pH i changes in WT or Nhe1 cKO microglia, we conducted ow cytometry with a novel pH indicator pHrodo whose uorescent intensity is inversely correlated with pH i (22) (Fig. 4a).…”

Section: Deletion Of Microglial Nhe1 Acidi Ed Microglia With Activate...mentioning

confidence: 99%

Deletion of Slc9a1 in Cx3cr1+ cells stimulated microglial subcluster CREB1-BDNF signaling and microglia-oligodendrocyte crosstalk

Song,

Oft,

Metwally

et al. 2023

Preprint

View full text Add to dashboard Cite

Microglial Na/H exchanger-1 (NHE1) protein, encoded by Slc9a1, plays a role in white matter demyelination of ischemic stroke brains. To explore underlying mechanisms, we conducted single cell RNA-seq transcriptome analysis in conditional Slc9a1 knockout (cKO) and wild-type (WT) mouse white matter tissues at 3 days post-stroke. Compared to WT, Nhe1 cKO brains expanded a microglial subgroup with elevated transcription of white matter myelination genes including Spp1, Lgals3, Gpnmb, and Fabp5. This subgroup also exhibited more acidic pHi and significantly upregulated CREB signaling and BDNF secretion detected by ingenuity pathway analysis and flow cytometry. Moreover, the Nhe1 cKO white matter tissues showed enrichment of a corresponding oligodendrocyte subgroup, with pro-phagocytosis and lactate shuffling gene expression, where activated CREB signaling is a likely upstream regulator. These findings demonstrate that attenuation of NHE1-mediated H+ extrusion acidifies microglia and stimulates microglial CREB1-BDNF signaling, giving rise to restorative microglia-oligodendrocyte interactions to potentially improve post-stroke remyelination.

show abstract

Protocol to record and quantify the intracellular pH in contracting cardiomyocytes

Cited by 2 publications

References 18 publications

Deletion of Slc9a1 in Cx3cr1+ cells stimulated microglial subcluster CREB1 signaling and microglia-oligodendrocyte crosstalk

Deletion of Slc9a1 in Cx3cr1+ cells stimulated microglial subcluster CREB1 signaling and microglia-oligodendrocyte crosstalk

Deletion of Slc9a1 in Cx3cr1+ cells stimulated microglial subcluster CREB1-BDNF signaling and microglia-oligodendrocyte crosstalk

Contact Info

Product

Resources

About