Coupled amplification and degradation of exogenous RNA injected in amphibian oocytes

AUTOR(ES)

Montreau, Nicole

FONTE

Oxford University Press

RESUMO

The early development of amphibians takes place in the absence of significant transcription and is controlled at the post-transcriptional level. We have reported that in vitro synthesized transcripts injected into axolotl fertilized eggs or oocytes were not continuously degraded as their abundance apparently fluctuated over time, with detected amounts sometimes higher than initial injected amounts. To further characterize this phenomenon, we have co-injected RNA chain terminators to prevent RNA synthesis. This led to the suppression of fluctuations and to a regular decrease in the amount of transcripts that appeared to be more stable in the presence of inhibitors. These observations indicate a coupling between RNA synthesis and an accelerated degradation. Throughout the time course, cRNA molecules could be detected, and their abundance increased in the early phase of the kinetics, supporting the implication of an RNA-dependent RNA polymerase in an asymmetric amplification process. Finally, when the fate of the injected transcripts was investigated in individual oocytes, we observed an absolute increase in abundance in some but not all oocytes, supporting the existence of a limiting step in the initiation of the RNA amplification stochastic process.

Documentos Relacionados

• In vivo repair of the 3'terminus of transfer RNA injected into amphibian oocytes.
• Microinjection of Rabbit Hemoglobin Messenger RNA into Amphibian Oocytes and Embryos
• Analysis of reconstruction of an RNP particle which stores 5S RNA and tRNA in amphibian oocytes.
• Differences in receptor-evoked membrane electrical responses in native and mRNA-injected Xenopus oocytes.
• Calcium channel currents in Xenopus oocytes injected with rat skeletal muscle RNA.