Methodological improvements for fluorescence recordings in Xenopus laevis oocytes.

ABSTRACT: Xenopus laevis oocytes are a widely used model system because of their capacity to translate exogenous mRNA, but their high intrinsic background fluorescence is a disadvantage for fluorescence recordings. Here, we developed two distinct methods for improving fluorescence recordings from oocytes. One was a pharmacological method in which a small-molecule salt-inducible kinase inhibitor was co-injected with the mRNA of interest to stimulate melanin production. We interrogated the oocytes using cut-open voltage clamp with simultaneous fluorescence recording and found that by increasing the amount of light-absorbing melanin in these oocytes, we decreased their intrinsic background fluorescence. The treated oocytes produced fluorescence signals that were approximately four times larger.

SUBMITTER: Lee EEL 

PROVIDER: S-EPMC6363413 | biostudies-literature | 2019 Feb

REPOSITORIES: biostudies-literature