Vacuolar Pathway
Vacuolar Pathway - 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).
01:27
C4 Pathway and CAM
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2019
Most plants use the C3 pathway for carbon fixation. However, some plants, such as sugar cane, corn, and cacti that grow in hot conditions, use alternative pathways to fix carbon and conserve energy loss due to photorespiration. Photorespiration is the process that occurs when the oxygen concentration is high. Under such conditions, the rubisco enzyme in the Calvin cycle binds O2 instead of CO2, which halts photosynthesis and consumes energy. C4 Pathway The C4 pathway is used by plants such as...
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.
FAQs
Cargo sorting determines whether material continues toward the vacuole after processing in the Golgi apparatus. The Golgi organizes cargo into vesicles, which provide a route through endosomes before delivery to the tonoplast. This step coordinates the movement of proteins, lipids, and other materials through the endomembrane system and supports their appropriate delivery to the vacuole.
Endosomes serve as an intermediate compartment between Golgi-derived vesicles and the tonoplast. Cargo passes through this stage before the transport route reaches the vacuole, linking sorting events in the Golgi with later membrane delivery. Their position in the pathway helps organize intracellular trafficking and contributes to the controlled movement of diverse materials.
Fusion with the tonoplast provides the final membrane-delivery step for cargo traveling through the pathway. Once this connection occurs, materials can reach the vacuole, where they contribute to storage, degradation, or cellular homeostasis. The process therefore connects intracellular transport with the functional roles of the vacuole in plant cells.
The pathway supports homeostasis by directing materials to a compartment involved in storage and degradation. It also contributes to nutrient storage, ion balance, and responses to environmental stress. Because these functions depend on the delivery of proteins, lipids, and other cargo, changes in trafficking can influence how plant cells maintain internal conditions and respond to their surroundings.
A conceptual workflow begins with cargo entering the endomembrane system at the endoplasmic reticulum. It then moves through the Golgi apparatus, where cargo is sorted into vesicles. These vesicles pass through endosomes and eventually reach the tonoplast, whose fusion with the transport route allows delivery to the vacuole. This sequence provides a framework for studying intracellular trafficking.
Studying this pathway helps researchers examine plant development, cargo sorting, nutrient storage, ion balance, and responses to environmental stress. It also provides context for biotechnology projects involving engineered storage proteins, because successful delivery to the vacuole is relevant to how such proteins are handled within plant cells. The pathway therefore connects cell biology with applied plant research.