Analysis of Nuclear Pore Complex Permeability in Mammalian Cells and Isolated Nuclei Using Fluorescent Dextrans

Area of Science:

• Cell Biology
• Molecular Biology
• Biophysics

Background:

• The nuclear envelope in eukaryotic cells separates the genome from the cytoplasm.
• Nuclear pore complexes (NPCs) are protein channels regulating molecule transport and forming a permeability barrier.
• Nuclear permeability can change with cell aging and disease.

Purpose of the Study:

• To provide a straightforward protocol for analyzing nuclear permeability.
• To enable researchers to study NPC function and nuclear transport dynamics.
• To investigate variations in nuclear permeability in different cellular contexts.

Main Methods:

• Utilizing plasma membrane-permeabilized cells and isolated nuclei.
• Employing fluorescently labeled dextrans of various sizes.
• Analyzing dextran diffusion using confocal microscopy.

Main Results:

• The protocol allows for quantitative assessment of nuclear permeability.
• Changes in nuclear pore complex barrier function can be detected.
• The method is applicable to diverse cell types and experimental conditions.

Conclusions:

• This protocol offers a valuable tool for studying nuclear pore complex function.
• It facilitates research into the dynamics of nuclear permeability in health and disease.
• The method supports investigations across various cell types and biological processes.