Fluorescent Tracking of the Endoplasmic Reticulum in Live Pathogenic Fungal Cells

Area of Science:

• Mycology
• Cell Biology
• Biochemistry

Background:

• The endoplasmic reticulum (ER) is crucial for ergosterol biosynthesis in fungi.
• Ergosterol is essential for fungal cell membranes and a target for azole antifungal drugs.
• Specific fluorescent labeling of the fungal ER is needed to study its function and drug interactions.

Purpose of the Study:

• To develop and evaluate novel fluorescent ER trackers for pathogenic fungi.
• To enable live-cell imaging of the ER in Candida species.
• To investigate ER-mitochondria interactions in pathogenic fungi.

Main Methods:

• Development of a collection of fluorescent ER trackers.
• Evaluation of trackers in a panel of Candida species.
• Confocal microscopy for live-cell imaging.
• Use of mitotracker in combination with ER trackers.

Main Results:

• Successfully developed fluorescent ER trackers for live fungal cell imaging.
• Demonstrated ER specificity linked to the azole drug target enzyme.
• Observed specific ER labeling in wild-type but not mutant Candida cells.
• Visualized ER-mitochondria contact sites using combined trackers.

Conclusions:

• Novel fluorescent ER trackers are effective tools for studying pathogenic fungi.
• These trackers facilitate research on the ER and its organelle interactions.
• The tools aid in understanding fungal cell biology and azole drug mechanisms.