Immunofluorescence of Microtubule Assemblies in Amphibian Oocytes and Early Embryos

Nguyen, Thao; Mitchison, Timothy J.; Wühr, Martin

doi:10.1007/978-1-4939-9009-2_2

Cited by 2 publications

(1 citation statement)

References 37 publications

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“…Immunofluorescence of HP1α and Lamin-A/C is referred to in Nguyen et al (2019) . After nuclear transfer, oocytes were fixed in low FG fixative and post-fixed with MeOH/EGTA.…”

Section: Methodsmentioning

confidence: 99%

Deterministic nuclear reprogramming of mammalian nuclei to a totipotency-like state by Amphibian meiotic oocytes for stem cell therapy in humans

Wen,

Barbosa Triana,

Butler

et al. 2024

Biology Open

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The ultimate aim of nuclear reprogramming is to provide stem cells or differentiated cells from unrelated cell types as a cell source for regenerative medicine. A popular route towards this is transcription factor induction, and an alternative way is an original procedure of transplanting a single somatic cell nucleus to an unfertilized egg. A third route is to transplant hundreds of cell nuclei into the germinal vesicle (GV) of a non-dividing Amphibian meiotic oocyte, which leads to the activation of silent genes in 24 hours and robustly induced a totipotency-like state in almost all transplanted cells. We apply the third route for potential therapeutic use and describe a procedure by which the differentiated states of cells can be reversed so that totipotency and pluripotency gene expression are regained. Differentiated cells are exposed to GV extracts and are reprogrammed to form embryoid bodies, which shows the maintenance of stemness and could be induced to follow new directions of differentiation. We conclude that much of the reprogramming effect of eggs is already present in meiotic oocytes and does not require cell division or selection of dividing cells. Reprogrammed cells by oocytes could serve as replacements for defective adult cells in humans.

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“…Immunofluorescence of HP1α and Lamin-A/C is referred to in Nguyen et al (2019) . After nuclear transfer, oocytes were fixed in low FG fixative and post-fixed with MeOH/EGTA.…”

Section: Methodsmentioning

confidence: 99%

Deterministic nuclear reprogramming of mammalian nuclei to a totipotency-like state by Amphibian meiotic oocytes for stem cell therapy in humans

Wen,

Barbosa Triana,

Butler

et al. 2024

Biology Open

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Differential nuclear import sets the timing of protein access to the embryonic genome

et al. 2022

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The development of a fertilized egg to an embryo requires the proper temporal control of gene expression. During cell differentiation, timing is often controlled via cascades of transcription factors (TFs). However, in early development, transcription is often inactive, and many TF levels stay constant, suggesting that alternative mechanisms govern the observed rapid and ordered onset of gene expression. Here, we find that in early embryonic development access of maternally deposited nuclear proteins to the genome is temporally ordered via importin affinities, thereby timing the expression of downstream targets. We quantify changes in the nuclear proteome during early development and find that nuclear proteins, such as TFs and RNA polymerases, enter the nucleus sequentially. Moreover, we find that the timing of nuclear proteins’ access to the genome corresponds to the timing of downstream gene activation. We show that the affinity of proteins to importin is a major determinant in the timing of protein entry into embryonic nuclei. Thus, we propose a mechanism by which embryos encode the timing of gene expression in early development via biochemical affinities. This process could be critical for embryos to organize themselves before deploying the regulatory cascades that control cell identities.

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Immunofluorescence of Microtubule Assemblies in Amphibian Oocytes and Early Embryos

Cited by 2 publications

References 37 publications

Deterministic nuclear reprogramming of mammalian nuclei to a totipotency-like state by Amphibian meiotic oocytes for stem cell therapy in humans

Deterministic nuclear reprogramming of mammalian nuclei to a totipotency-like state by Amphibian meiotic oocytes for stem cell therapy in humans

Differential nuclear import sets the timing of protein access to the embryonic genome

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