Laird Diana J, Weissman Irving L
Department of Biological Sciences, Hopkins Marine Station, Stanford University, Pacific Grove, CA 93950, USA.
Dev Biol. 2004 Sep 15;273(2):185-94. doi: 10.1016/j.ydbio.2004.05.029.
Telomerase is critical for the protection of germ line and stem cell chromosomes from fatal shortening during replication. In most organisms, telomerase activity is suppressed in progressively committed cells and falls to basal rates in terminally differentiated lineages. The colonial ascidian Botryllus schlosseri propagates asexually and sexually, presumably from pools of stem cells that self-renew throughout the 2- to 5-year colony life span. Asexual budding takes place continuously from the parental body wall. When the colony reaches a critical size, sexual reproduction commences with the generation of gonads. Here, we establish the existence of 6-15 kb telomeres on the ends of Botryllus chromosomes. We develop a real-time quantitative PCR telomeric repeat amplification protocol (TRAP) assay that reliably detects 0.2-100 TPG units in cells and tissues. We find highest levels of enzymatic activity in the gonads, developing embryos, and tissues containing the earliest asexual buds. Telomerase activity appears to be suppressed in later buds during organogenesis and falls to basal rates in mature zooids. We postulate that this pattern reflects maximum telomere restoration in somatic stem cells of early buds and suppression of telomerase activity in progenitors and terminally differentiated cells, indicative of an alternate role for stem cells as repeated body regenerators in colonial life histories.
端粒酶对于保护生殖系和干细胞染色体在复制过程中免于致命性缩短至关重要。在大多数生物体中,端粒酶活性在逐渐分化的细胞中受到抑制,并在终末分化谱系中降至基础水平。群体海鞘Botryllus schlosseri通过无性和有性方式繁殖,推测源自干细胞池,这些干细胞在2至5年的群体寿命中进行自我更新。无性出芽持续发生于亲本体壁。当群体达到临界大小时,随着性腺的产生开始有性生殖。在此,我们确定了Botryllus染色体末端存在6 - 15 kb的端粒。我们开发了一种实时定量PCR端粒重复序列扩增协议(TRAP)检测方法,可可靠地检测细胞和组织中0.2 - 100 TPG单位。我们在性腺、发育中的胚胎以及含有最早无性芽的组织中发现了最高水平的酶活性。在器官发生过程中,端粒酶活性在后期芽中似乎受到抑制,并在成熟个体中降至基础水平。我们推测这种模式反映了早期芽的体细胞干细胞中端粒的最大程度恢复以及祖细胞和终末分化细胞中端粒酶活性的抑制,这表明干细胞在群体生活史中作为反复身体再生者具有另一种作用。