TY  - GEN
T1  - Mutations linked to loss of cell cycle control can render cells responsive to local differentiation cues
AU  - Cerveny, Kara L
AU  - Bronstein, Hannah
AU  - Hagen, Olivia
AU  - Lamb, Dayna B
AU  - Martin, Grace
AU  - Tower, Ingrid
AU  - Van Duzer, Avery
AU  - Welch, Evan
AU  - Varga, Máté
DO  - 10.17912/micropub.biology.000481
UR  - http://beta.micropublication.org/journals/biology/micropub-biology-000481/
AB  - Cell behaviors such as survival, proliferation, and death are governed by a bevy of cues, both from within the cells themselves and from the local tissue environment. To test whether a wild-type environment could encourage the survival and/or differentiation of neuronal progenitor cells with impaired cell cycle progression, we created chimeric zebrafish embryos containing mutant cells in wild-type retinae. We transplanted 10-20 cells from blastula stage donors into the region of early gastrula stage wild-type hosts fated to be eye field. Donor and host embryo pairs were cultured together; donors were genotyped when possible at 1 day post fertilization (dpf). We screened all transplants at 1 dpf for survival and location of clones. Hosts that contained labeled clones in their eyes were fixed at 3 dpf and either cryosectioned before immunostaining or subjected to immunostaining as wholemounts. All embryos analyzed in this study were imaged with epifluorescence and/or confocal microscopy. Consistent with previous reports, we found that cdk1, dtl, slbp, fbxo5, ahctf1, gins2, hdac1, mcm5, ssrp1a, and rbbp6 mutant retinae contained dying cells with pyknotic nuclei throughout the developing retinal neuroepithelium at 3 dpf. Moreover, all of these mutant embryos had eyes that were noticeably smaller than their wild-type siblings (Table 1; references therein). When cells from mutant embryos were transplanted into wild-type hosts, survival and/or differentiation was almost always compromised in a manner consistent with cell-autonomous cell death. In particular, wild-type hosts that contained clones of cdk1, dtl, mcm5, and rbbp6 mutant cells at 1 dpf rarely contained visible clones by 3 dpf (see Table 1 for numbers of chimeras analyzed and how many clones survived until 3 dpf ). For those clones that were visible, they were small (e.g., compare Fig 1A and 1F) and/or exhibited signs of apoptosis (e.g. Fig 1H). For example, by transplanting gins2 morphant cells labeled with a membrane-targeted red fluorescent protein into wild-type embryos, we observed mutant cells blebbing and/or fragmenting when integrated into wild-type retinae (Fig 1H) whereas wild-type sibling cells integrated fully into the host environment, highlighting typical retinal neuronal morphologies (Fig 1G).
PY  - 2021
JO  - microPublication Biology
ER  -