Vacuolar Ph
Vacuolar Ph - Related Videos
09:25
Quantification of Cytosolic vs. Vacuolar Salmonella in Primary Macrophages by Differential Permeabilization
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Cited by 19 •
2015
We describe the quantification of cytosolic and vacuolar Salmonella typhimurium in bone-marrow derived macrophages using differential digitonin permeabilization.
06:27
An Assay to Quantify Cytosolic and Vacuolar Salmonella in Infected Primary Macrophages
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2025
This video demonstrates an assay to quantify cytosolic and vacuolar Salmonella typhimurium inside bone marrow-derived macrophages using differential permeabilization. The macrophages are infected with a fluorescent protein-expressing Salmonella, which is internalized inside a specialized vacuolar structure called Salmonella-containing vacuole or SCV. A subset of bacteria escape the SCV and enter the cytosol. The cholesterol-rich plasma membrane of macrophages is permeabilized using digitonin,...
13:55
Measurement of Vacuolar and Cytosolic pH In Vivo in Yeast Cell Suspensions
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Cited by 40 •
2013
Vacuolar and cytosolic pH can be measured in live yeast (S. cerevisiae) cells using ratiometric fluorescent dyes localized to specific cellular compartments. We describe procedures for measuring vacuolar pH with BCECF-AM, which localizes to the vacuole in yeast, and cytosolic pH with a cytosolic ratiometric pH-sensitive GFP (yeast pHluorin).
10:39
Single Cell Measurements of Vacuolar Rupture Caused by Intracellular Pathogens
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Cited by 22 •
2013
We describe a method for tracking the endomembrane rupture elicited by the intracellular bacteria Shigella flexneri and Mycobacterium tuberculosis upon host cell invasion. Our assay makes use of CCF4, a host cytoplasmic FRET probe in live or fixed cells. This reporter is degraded by an enzyme activity present on the bacterial surface.
02:52
A Technique to Establish a Bacterial Infection Model in an Ex Vivo Organ Culture
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2025
This video demonstrates a method to establish an infection model in placental tissues to study infection propagation. An ex vivo placental organ culture is infected with Listeria monocytogenes, a facultative intracellular parasite. The bacteria invade extravillous trophoblasts (EVTs) to enter the placental tissue and then propagate to other cells via cell-to-cell spread.
FAQs
Vacuolar H+-ATPases use proton transport across the vacuolar membrane to build an electrochemical gradient, meaning both hydrogen-ion concentration and electrical charge differ across the membrane. This gradient supplies the underlying force for maintaining the vacuole’s acidic internal conditions. Its stability depends on coordinated membrane transport rather than proton pumping alone, linking acidity to broader cellular homeostasis.
Moving hydrogen ions creates charge and concentration differences that must remain balanced for the gradient to persist. Counterion transport and ion exchange help stabilize these changes across the vacuolar membrane, preventing proton movement from becoming self-limiting. Their contribution explains why vacuolar acidity depends on a network of coordinated ionic processes instead of a single proton-transport mechanism.
Changes in vacuolar pH can affect processes that depend on the organelle’s acidic environment, including molecular storage, waste processing, protein degradation, and membrane trafficking. Because these activities contribute to cellular homeostasis, disrupted acidity can extend beyond the vacuole itself. Studying such changes helps connect organelle-level ion regulation with altered cellular function and disease-related processes.
Researchers can examine vacuolar pH directly as a way to investigate how organelle acidity relates to cellular function, or manipulate it to study the consequences of altered acidity. These approaches help distinguish whether changes in storage, degradation, trafficking, or stress responses accompany modified vacuolar conditions. The resulting observations clarify the functional importance of pH regulation within cells.
In plants, vacuolar pH provides context for understanding how the vacuole supports nutrient storage. Measuring or changing this property allows researchers to examine how acidity relates to the storage environment and to broader cellular homeostasis. This application places proton transport and ion balancing within a plant-specific framework, connecting membrane regulation with the biological handling of stored materials.
Vacuolar acidity is relevant because the vacuole participates in both membrane trafficking and protein degradation. Investigating its pH helps researchers determine how altered organelle conditions relate to these processes and whether cellular transport or breakdown functions change when acidity is disturbed. This provides a way to study the connection between membrane compartments, molecular turnover, and overall cell performance.