INTERRELATIONSHIP BETWEEN POLY(ADP-RIB) SYNTHESIS, INTRACELLULAR NAD LEVELS, AND MUSCLE OR CARTILAGE DIFFERENTIATION FROM MESODERMAL CELLS OF EMBRYONIC CHICK LIMB
INTERRELATIONSHIP BETWEEN POLY(ADP-RIB) SYNTHESIS, INTRACELLULAR NAD LEVELS, AND MUSCLE OR CARTILAGE DIFFERENTIATION FROM MESODERMAL CELLS OF EMBRYONIC CHICK LIMB
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DOI：
10.1073/pnas.72.5.1852
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发表时间：
1975-01-01
影响因子：
11.1
通讯作者：
ROSENBERG, MJ
中科院分区：
文献类型：
--
作者：
CAPLAN, AI;ROSENBERG, MJ
鸡胚肢体的中胚层细胞具有分化为肌肉或软骨的能力。来自该实验室的先前报告显示了吡啶核苷酸水平与这种分化之间的相关性，因此表明细胞NAD水平的波动在控制肌肉与软骨发育中起作用。Poly（ADP-Rib）是染色质相关的，并且由NAD的聚合与烟酰胺的切除形成。本文报道的研究表明：（A）聚（ADP-Rib）的净合成速率与体外培养的第24期肢体中胚层细胞向软骨细胞分化有关;（B）由暴露于烟酰胺或BrdUrd引起的软骨形成表达的抑制与聚（ADP-Rib B）合成的基础水平的维持相关，并且这种抑制依赖于外源性提供的烟酰胺或BrdUrd的浓度;（C）体外肢体中胚层细胞暴露于3-乙酰吡啶引起的软骨形成表达的增强与聚（ADP-Rib）如果针对ATP库的比活性校正或与未处理的培养物相比，基于每个细胞的合成;（D）来自中胚层细胞的分离的染色质具有用于合成聚（ADP-Rib）的酶机制;（E）该机制被烟酰胺、胸苷和3-乙酰吡啶抑制;和（F）新合成的聚（ADP-Rib）与染色质的离散部分结合或通过高柱盐从染色质中完全提取，这导致聚集，最终在琼脂糖柱的排阻体积位置洗脱。总之，这些观察结果为细胞NAD水平的波动如何与基因组相关机制进行通信或被其“感知”并最终导致差异表型表达提供了可能的解释。
Mesodermal cells of embryonic chick limbs have the capacity to differentiate into either muscle or cartilage. Previous reports from this laboratory show a correlation between pyridine nucleotide levels and this differentiation, and thus suggest that fluctuations in the cellular NAD levels play a role in the control of muscle versus cartilage development. Poly(ADP-Rib) is chromatin-associated and forms from the polymerization of NAD with the excision of nicotinamide. The studies reported here show that: (A) the rate of net synthesis of poly(ADP-Rib) is correlated with the differentiation of chondrogenic cells from stage 24 limb mesodermal cells grown in vitro; (B) inhibition of chondrogenic expression caused by exposure to nicotinamide or BrdUrd is correlated with maintenance of basal levels of poly(ADP-Rib) synthesis, and this inhibition is dependent on the concentration of nicotinamide or BrdUrd exogenously supplied; (C) potentiation of chondrogenic expression caused by exposure of limb mesodermal cells in vitro to 3-acetylpyridine is correlated with stimulation of the rate of poly(ADP-Rib) synthesis if corrected for the specific activity of the ATP pool or compared to untreated cultures on a per cell basis; (D) isolated chromatin from mesodermal cells has the enzymatic machinery for synthesizing poly(ADP-Rib); (E) this machinery is inhibited by nicotinamide, thymidine, and 3-acetylpyridine; and (F) newly synthesized poly(ADP-Rib) is either associated with a discrete fraction of chromatin or is completely extracted from chromatin by the high column salts, which result in an aggregation with eventual elution at the exclusion volume position of the agarose column. Taken together, these observations provide a possible explanation for how fluctuations in cellular NAD levels can communicate with or be "sensed" by genomic related machinery and eventually result in differtial phenotypic expression.