Biology subjects

Genin, O.

Publications and source records attributed to Genin, O..

3 recordsLinked to original sources

bioRxiv2022

HREM, RNAseq and cell-cycle analyses reveal the role of the G2/M-regulatory protein, Wee1, on the survivability of chicken embryos during diapause.

Avian blastoderm can enter into diapause when kept at low temperatures, and successfully resume development (SRD) when re-incubated in body-temperature. These abilities, which are largely affected by the temperature and duration of the diapause, are poorly understood at the cellular and molecular level. To determine how temperature affects embryonic morphology during diapause, High-Resolution Episcopic Microscopy (HREM) analysis was utilized. While blastoderms diapausing at 12{degrees}C for 28 days presented typical cytoarchitecture, similar to non-diapaused embryos, at 18{degrees}C much thicker blastoderms with higher cell-number were observed. RNAseq was conducted to discover the genes underlying these phenotypes, revealing differentially-expressed cell-cycle regulatory genes. Amongst them, Wee1, a negative-regulator of G2/M transition, was highly expressed at 12{degrees}C compared to 18{degrees}C. This finding suggested that cells at 12{degrees}C are arrested at the G2/M phase, as supported by bromodeoxyuridine incorporation (BrdU) assay and phosho-histone-H3 (pH3) immuno-staining. Inhibition of Wee1 during diapause at 12{degrees}C resulted in cell-cycle progression beyond the G2/M and augmented tissue volume, resembling the morphology of 18{degrees}C-diapaused embryos. These findings suggest that diapause at low temperatures leads to Wee1 upregulation which arrests the cell-cycle at the G2/M phase, promoting the perseverance of embryonic cytoarchitecture and future SRD. In contrast, Wee1 is not up-regulated during diapause at higher temperature, leading to continuous proliferation and maladaptive morphology associated with poor survivability. Combining HREM-based analysis with RNAseq and molecular manipulations, we present a novel mechanism that regulates the ability of diapaused-avian embryos to maintain their cytoarchitecture via cell-cycle arrest, which enables their SRD.

evolutionary biology↗

bioRxiv2021

How do avian embryos resume development following diapause? A new role for TGF-β in regulating pluripotency-related genes

Avian embryos can halt their development for long periods at low temperature in a process called diapause and successfully resume development when reincubated at maternal body temperature. Successful resumption of development depends on different factors, including temperature. We have recently shown that embryos that enter diapause at 18 {degrees}C present a significant reduction in their ability to develop normally when put back into incubation, compared to embryos entering diapause at 12 {degrees}C. However, the mechanisms underlying these differences are unknown. To address this question, transcriptome analysis was performed to compare the effect of diapause temperature on gene expression, and to identify pathways involved in the process. Genetic comparison and pathway-enrichment analysis revealed that TGF-{beta} and pluripotency-related pathways are differentially regulated at the two temperatures, with higher expression at 12 {degrees}C compared to 18 {degrees}C. Investigating the involvement of the TGF-{beta} pathway revealed an essential role for BMP4 in regulating the expression of the transcription factors Nanog and Id2, which are known to regulate pluripotency and self-renewal in embryonic stem cells. BMP4 gain- and loss-of-function experiments in embryos in diapause at the different temperatures revealed the main role of BMP4 in enabling resumption of normal development following diapause. Collectively, these findings identify molecular regulators that facilitate embryos ability to undergo diapause at different temperatures and resume a normal developmental program.

developmental biology↗

bioRxiv2019

Early pathological signs in young dysf-/- mice are improved by halofuginone

Dysferlinopathies are a non-lethal group of late-onset muscular dystrophies. Here, we evaluated the fusion ability of primary myoblasts from young dysf-/- mice and the muscle histopathology prior to, and during early stages of disease onset. The ability of primary myoblasts of 5-weekold dysf-/- mice to form large myotubes was delayed compared to their wild-type counterparts, as evaluated by scanning electron microscopy. However, their fusion activity, as reflected by the presence of actin filaments connecting several cells, was enhanced by the antifibrotic drug halofuginone. Early dystrophic signs were already apparent in 4-week-old dysf-/- mice; their collagen level was double that in wild-type mice and continued to rise until 5 months of age. Continuous treatment with halofuginone from 4 weeks to 5 months of age reduced muscle fibrosis in a phosphorylated-Smad3 inhibition-related manner. Halofuginone also enhanced myofiber hypertrophy, reduced the percentage of centrally nucleated myofibers, and increased muscle performance. Together, the data suggest an inhibitory effect of halofuginone on the muscle histopathology at very early stages of dysferlinopathy, and better generation of force and muscle performance. These results offer new opportunities for early pharmaceutical treatment in dysferlinopathies with favorable outcomes at later stages of life.

cell biology↗