ArticleEukaryotic cell2009
Live-Cell imaging and measurement of intracellular pH in filamentous fungi using a genetically encoded ratiometric probe.
Article in Eukaryotic cell, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 31 papers.
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Who cites it
31 citing papers in PubMed, 86 citations in OpenAlex.
- AlphaFold modeling uncovers global structural features of class I and class II fungal hydrophobins.Protein science : a publication of the Protein Society · 2025Article
- Hyphae of the fungus Aspergillus nidulans demonstrate chemotropism to nutrients and pH.PLoS biology · 2024Article
- Unlocking the distinctive enzymatic functions of the early plant biomass deconstructive genes in a brown rot fungus by cell-free protein expression.Applied and environmental microbiology · 2024Article
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- Novel Identification of the Collection of Pathogenic Fungal SpeciesPathogens (Basel, Switzerland) · 2023Article
- Enhancing biofuels production by engineering the actin cytoskeleton in Saccharomyces cerevisiae.Nature communications · 2022Article
- Generating new mixtures of food additives with antimicrobial and cytotoxic potency againstFood science & nutrition · 2022Article
- Advances and Challenges in FluorescenceFrontiers in microbiology · 2022Article
- Bioleaching of Manganese Oxides at Different Oxidation States by Filamentous FungusJournal of fungi (Basel, Switzerland) · 2021Article
- Virtual 2-D map of the fungal proteome.Scientific reports · 2021Article
- Establishment of the monomeric yellow-green fluorescent protein mNeonGreen for life cell imaging in mycelial fungi.AMB Express · 2020Article
- Unconventional Secretion of Nigerolysins A fromMicroorganisms · 2020Article
- Differential Effect of Extracellular Acidic Environment on IL-1β Released from Human and Mouse Phagocytes.International journal of molecular sciences · 2020Article
- Tolerance to alkaline ambient pH in Aspergillus nidulans depends on the activity of ENA proteins.Scientific reports · 2020Article
- Self-assembled amphiphilic fluorescent probe: detecting pH-fluctuations within cancer cells and tumour tissues.Chemical science · 2020Article
- Transient intracellular acidification regulates the core transcriptional heat shock response.eLife · 2020Article
- Live Cell Measurement of the Intracellular pH of Yeast by Flow Cytometry Using a Genetically-Encoded Fluorescent Reporter.Bio-protocol · 2020Article
- Regulators of nitric oxide signaling triggered by host perception in a plant pathogen.Proceedings of the National Academy of Sciences of the United States of America · 2020Article
- Article
- Fluorescent Proteins for Investigating Biological Events in Acidic Environments.International journal of molecular sciences · 2018Review
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Authors and funding
4 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
A novel, genetically encoded, ratiometric pH probe (RaVC) was constructed to image and measure intracellular pH in living hyphae of Aspergillus niger. RaVC is a chimeric protein based on the pH-sensitive probe pHluorin, which was partially codon optimized for expression in Aspergillus. Intracellular pH imaging and measurement was performed by simultaneous, dual-excitation confocal ratio imaging. The mean cytoplasmic pH measured was 7.4 to 7.7 based on calibrating RaVC in situ within nigericin-treated hyphae. Pronounced, longitudinal cytoplasmic pH gradients were not observed in the apical 20 microm of actively growing hyphae at the periphery of 18-h-old colonies. The cytoplasmic pH remained unchanged after prolonged growth in buffered medium with pH values between 2.5 or 9.5. Sudden changes in external pH significantly changed cytoplasmic pH by <1.3 pH units, but it returned to its original value within 20 min following treatment. The weak acid and antifungal food preservative sorbic acid caused prolonged, concentration-dependent intracellular acidification. The inhibition of ATPases with N-ethylmaleimide, dicychlohexylcarbodimide, or sodium azide caused the cytoplasmic pH to decrease by <1 pH unit. Treatment with the protonophore carbonyl cyanide m-chlorophenylhydrazone or cyanide p-(trifluoromethoxy) phenylhydrazone reduced the cytoplasmic pH by <1 pH unit. In older hyphae from 32-h-old cultures, RaVC became sequestered within large vacuoles, which were shown to have pH values between 6.2 and 6.5. Overall, our study demonstrates that RaVC is an excellent probe for visualizing and quantifying intracellular pH in living fungal hyphae.
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