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1.
Dev Biol ; 373(1): 196-204, 2013 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-23103585

RESUMO

Limb regeneration involves re-establishing a limb development program from cells within adult tissues. Identifying molecular handles that provide insight into the relationship between cell differentiation status and cell lineage is an important step to study limb blastema cell formation. Here, using single cell PCR, focusing on newly isolated Twist1 sequences, we molecularly profile axolotl limb blastema cells using several progenitor cell markers. We link their molecular expression profile to their embryonic lineage via cell tracking experiments. We use in situ hybridization to determine the spatial localization and extent of overlap of different markers and cell types. Finally, we show by single cell PCR that the mature axolotl limb harbors a small but significant population of Twist1(+) cells.


Assuntos
Ambystoma mexicanum/fisiologia , Tecido Conjuntivo/metabolismo , Extremidades/fisiologia , Músculo Esquelético/metabolismo , Regeneração/fisiologia , Células-Tronco/metabolismo , Proteína 1 Relacionada a Twist/metabolismo , Animais , Linhagem da Célula/fisiologia , Células do Tecido Conjuntivo/metabolismo , Hibridização In Situ , Mesoderma/citologia , Músculo Esquelético/citologia , Reação em Cadeia da Polimerase , Pele/citologia , Transcriptoma
2.
J Biophotonics ; 14(4): e202000457, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33345429

RESUMO

Transparency is widespread in nature, ranging from transparent insect wings to ocular tissues that enable you to read this text, and transparent marine vertebrates. And yet, cells and tissue models in biology are usually strongly light scattering and optically opaque, precluding deep optical microscopy. Here we describe the directed evolution of cultured mammalian cells toward increased transparency. We find that mutations greatly diversify the optical phenotype of Chinese Hamster Ovary cells, a cultured mammalian cell line. Furthermore, only three rounds of high-throughput optical selection and competitive growth are required to yield fit cells with greatly improved transparency. Based on 15 monoclonal cell lines derived from this directed evolution experiment, we find that the evolved transparency frequently goes along with a reduction of nuclear granularity and physiological shifts in gene expression profiles. In the future this optical plasticity of mammalian cells may facilitate genetic clearance of living tissues for in vivo microscopy.


Assuntos
Microscopia , Animais , Células CHO , Cricetinae , Cricetulus
3.
Science ; 347(6229): 1465-70, 2015 Mar 27.
Artigo em Inglês | MEDLINE | ID: mdl-25721503

RESUMO

Evolutionary expansion of the human neocortex reflects increased amplification of basal progenitors in the subventricular zone, producing more neurons during fetal corticogenesis. In this work, we analyze the transcriptomes of distinct progenitor subpopulations isolated by a cell polarity-based approach from developing mouse and human neocortex. We identify 56 genes preferentially expressed in human apical and basal radial glia that lack mouse orthologs. Among these, ARHGAP11B has the highest degree of radial glia-specific expression. ARHGAP11B arose from partial duplication of ARHGAP11A (which encodes a Rho guanosine triphosphatase-activating protein) on the human lineage after separation from the chimpanzee lineage. Expression of ARHGAP11B in embryonic mouse neocortex promotes basal progenitor generation and self-renewal and can increase cortical plate area and induce gyrification. Hence, ARHGAP11B may have contributed to evolutionary expansion of human neocortex.


Assuntos
Proteínas Ativadoras de GTPase/fisiologia , Regulação da Expressão Gênica no Desenvolvimento , Neocórtex/embriologia , Células-Tronco Neurais/citologia , Neurogênese/genética , Animais , Separação Celular , Proteínas Ativadoras de GTPase/química , Proteínas Ativadoras de GTPase/genética , Duplicação Gênica , Humanos , Ventrículos Laterais/citologia , Camundongos , Neocórtex/citologia , Neocórtex/metabolismo , Células-Tronco Neurais/metabolismo , Neuroglia/citologia , Neuroglia/metabolismo , Neurônios/citologia , Neurônios/metabolismo , Estrutura Terciária de Proteína , Transcriptoma
4.
Nat Commun ; 2: 154, 2011 Jan 11.
Artigo em Inglês | MEDLINE | ID: mdl-21224845

RESUMO

During mammalian cerebral cortex development, the G1-phase of the cell cycle is known to lengthen, but it has been unclear which neural stem and progenitor cells are affected. In this paper, we develop a novel approach to determine cell-cycle parameters in specific classes of neural stem and progenitor cells, identified by molecular markers rather than location. We found that G1 lengthening was associated with the transition from stem cell-like apical progenitors to fate-restricted basal (intermediate) progenitors. Unexpectedly, expanding apical and basal progenitors exhibit a substantially longer S-phase than apical and basal progenitors committed to neuron production. Comparative genome-wide gene expression analysis of expanding versus committed progenitor cells revealed changes in key factors of cell-cycle regulation, DNA replication and repair and chromatin remodelling. Our findings suggest that expanding neural stem and progenitor cells invest more time during S-phase into quality control of replicated DNA than those committed to neuron production.

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