Roy Debasmita, Michaelson David, Hochman Tsivia, Santella Anthony, Bao Zhirong, Goldberg Judith D, Hubbard E Jane Albert
Skirball Institute of Biomolecular Medicine, Helen L. and Martin S. Kimmel Center for Stem Cell Biology, Departments of Cell Biology and Pathology, New York University School of Medicine, 540 First Avenue, New York, NY 10016, USA.
Departments of Population Health and Environmental Medicine, Division of Biostatistics, New York University School of Medicine, 540 First Avenue, New York, NY, 10016, USA.
Dev Biol. 2016 Jan 1;409(1):261-271. doi: 10.1016/j.ydbio.2015.10.031. Epub 2015 Nov 11.
Many organisms accumulate a pool of germline stem cells during development that is maintained in later life. The dynamics of establishment, expansion and homeostatic maintenance of this pool are subject to both developmental and physiological influences including the availability of a suitable niche microenvironment, nutritional status, and age. Here, we investigated the dynamics of germline proliferation during stages of expansion and homeostasis, using the C. elegans germ line as a model. The vast majority of germ cells in the proliferative zone are in interphase stages of mitosis (G1, S, G2) rather than in the active mitotic (M) phase. We examined mitotic index and DNA content, comparing different life stages, mutants, and physiological conditions. We found that germ cells in larval stages cycle faster than in adult stages, but that this difference could not be attributed to sexual fate of the germ cells. We also found that larval germ cells exhibit a lower average DNA content compared to adult germ cells. We extended our analysis to consider the effects of distance from the niche and further found that the spatial pattern of DNA content differs between larval and adult stages in the wild type and among mutants in pathways that interfere with cell cycle progression, cell fate, or both. Finally, we characterized expansion of the proliferative pool of germ cells during adulthood, using a regeneration paradigm (ARD recovery) in which animals are starved and re-fed. We compared adult stage regeneration and larval stage expansion, and found that the adult germ line is capable of rapid accumulation but does not sustain a larval-level mitotic index nor does it recapitulate the larval pattern of DNA content. The regenerated germ line does not reach the number of proliferative zone nuclei seen in the continuously fed adult. Taken together, our results suggest that cell cycle dynamics are under multiple influences including distance from the niche, age and/or maturation of the germ line, nutrition and, possibly, latitude for physical expansion.
许多生物体在发育过程中积累了一批生殖系干细胞库,并在随后的生命中得以维持。该干细胞库的建立、扩增和稳态维持动态受到发育和生理因素的影响,包括合适的小生境微环境的可用性、营养状况和年龄。在这里,我们以秀丽隐杆线虫的生殖系为模型,研究了生殖系在扩增和稳态阶段的增殖动态。增殖区的绝大多数生殖细胞处于有丝分裂的间期阶段(G1期、S期、G2期),而非活跃的有丝分裂(M)期。我们检测了有丝分裂指数和DNA含量,比较了不同生命阶段、突变体和生理条件。我们发现幼虫阶段的生殖细胞比成虫阶段的细胞周期更快,但这种差异不能归因于生殖细胞的性命运。我们还发现,与成虫生殖细胞相比,幼虫生殖细胞的平均DNA含量较低。我们进一步扩展分析,考虑到与小生境距离的影响,还发现野生型幼虫和成虫阶段以及干扰细胞周期进程、细胞命运或两者的途径中的突变体之间,DNA含量的空间模式存在差异。最后,我们利用再生范式(ARD恢复)对成虫期生殖细胞增殖库的扩增进行了表征,在该范式中,动物先饥饿后再喂食。我们比较了成虫期再生和幼虫期扩增,发现成虫生殖系能够快速积累,但无法维持幼虫水平的有丝分裂指数,也不能重现幼虫期的DNA含量模式。再生的生殖系无法达到持续喂食的成虫中增殖区细胞核的数量。综上所述,我们的结果表明,细胞周期动态受到多种影响,包括与小生境的距离、生殖系的年龄和/或成熟度、营养以及可能的物理扩张空间。