Logo image
Back
Component analysis of nucleolar protein compartments using Xenopus laevis oocytes
Journal article   Open access   Peer reviewed

Component analysis of nucleolar protein compartments using Xenopus laevis oocytes

Emily D Lavering, Irini N Petros and Daniel L Weeks
Development, growth & differentiation, Vol.64(6), pp.306-317
05/24/2022
DOI: 10.1111/dgd.12794
PMCID: PMC9474603
PMID: 35607824
Appears in  UI Libraries Support Open Access
url
https://doi.org/10.1111/dgd.12794View
Published (Version of record)CC BY-NC-ND V4.0,  Open Access

Abstract

The nucleolus is a multi-compartment, non-membrane-bound organelle within the nucleus. Nucleolar assembly is influenced by proteins capable of phase separation. Xenopus laevis oocytes contain hundreds of large nucleoli that provide experimental access for nucleoli that is unavailable in other systems. Here we detail methods to streamline the in vivo analysis of the compartmentalization of nucleolar proteins that are suspected of phase separation. The nucleolus is the main hub of ribosome biogenesis and here we present data supporting the division of proteins into nucleolar domains based on their function in ribosome biogenesis. We also describe the use of vital dyes such as Hoechst 33342 and Thioflavin T in nucleolar staining. Additionally, we quantify nucleolar morphology changes induced by heat shock and actinomycin D treatments. We suggest these approaches will be valuable in a variety of studies that seek to better understand the nucleolus, particularly those regarding phase separation. These approaches may also be instructive for other studies on phase separation, especially in the nucleus.
 cell nucleolus oocytes fluorescence microscopy Xenopus laevis biomolecular condensates UIOWA OA Agreement

Details

Metrics

24 Record Views
5 Times Cited - Web of Science
Logo image