ENHANCING USE OF SEA URCHIN EGGS AS A RESEARCH RESOURCE

这项工作的长期目标是提高海胆 配子系统作为细胞和发育研究的模型。拟议 研究将开发出体外储存排卵卵的程序， 延长时间，从两周到四周或更长时间， 保持受精能力和正常发育。这将使 实验依赖于在鸡蛋中预先加载同位素以达到平衡， 允许表达注射的mRNA，并允许对细胞进行多次实验。 同一批鸡蛋。最后，拥有长寿的卵子将使发育 运输卵子的方法（与运输难以维持的成虫相反）， 这将扩大使用这一资源的研究人员的数量。 海胆卵通常在卵巢中储存数月。我们 初步结果表明， 在无菌条件下培养排卵卵的寿命。因为鸡蛋 在卵巢中存活数月，我们的实验方法将 包括模拟卵巢状况拟议的研究将 检查这些参数，如低O2张力，高CO2， 营养素和抗氧化剂。当最佳条件已经 确定后，该程序将扩大规模，以处理大量的鸡蛋 材料 利用容易研究的海胆配子进行的调查提供了 对受精、细胞分裂和调节 通过转录因子的基因活性。这些发现导致 在脊椎动物中发现了类似的现象， 控制细胞周期素（cyclins），钙释放的替代手段 （环ADP核糖）和细胞内pH调节（Na+-H+交换）， 举几个例子。加强这一资源将扩大对这些问题的研究， 形式，从而增加了我们对一般细胞的知识， 所有动物的发育现象。

The long term objective of this work is the enhancement of the sea urchin gamete system as a model for cell and developmental studies. The proposed research will develop procedures to store ovulated eggs in vitro for extended periods of time, from two to four weeks or longer, and still maintain fertilizability and normal development. This would permit experiments that depend on preloading eggs with isotopes to equilibrium, allow expression of injected mRNA and permit multiple experiments on the same batch of eggs. Finally, having long-lived eggs will allow development of methods to ship eggs (as opposed to shipping hard-to-maintain adults), which would expand the pool of researchers using this resource. Sea urchin eggs are normally stored in the ovary for many months. Our preliminary results have demonstrated a significant extension of the lifetime of ovulated eggs by culture under axenic conditions. Because eggs remain viable for many months in the ovary, our experimental approach will involve mimicking the ovarian conditions. The proposed research will examine such parameters as low O2 tension, high CO2, provision of nutrients and antioxidants. When the optimal conditions have been determined, the procedure will be scaled to handle large amounts of egg material. Investigations using the easily studied sea urchin gametes have provided important insights into fertilization, cell division and regulation of gene activity through transcription factors. These findings have led to discovery of similar phenomena in vertebrates such as molecules that control call division (the cyclins), alternate means of calcium release (cyclic ADP ribose) and intracellular pH regulation (Na+-H+ exchangers) to name a few. Enhancement of this resource will expand research on these forms with a resultant increase in our knowledge of general cell and developmental phenomena applicable to all animals.