THE LIGAND OF THE C-KIT RECEPTOR PROMOTES OOCYTE GROWTH
THE LIGAND OF THE C-KIT RECEPTOR PROMOTES OOCYTE GROWTH
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DOI：
10.1006/dbio.1994.1020
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发表时间：
1994-01-01
影响因子：
2.7
通讯作者：
BACHVAROVA, RF
中科院分区：
文献类型：
--
作者：
PACKER, AI;HSU, YC;BACHVAROVA, RF
遗传学和描述性研究都表明c-kit受体及其配体KL参与了出生后小鼠卵巢中卵母细胞的生长过程。为了验证KL是一种卵母细胞生长因子的假设，我们利用卵母细胞培养系统研究了KL的体外效应。初步实验证实卵巢c-kit和KL都具有生物活性。免疫复合物激酶测定表明，主要存在于卵母细胞上的卵巢c-kit具有自磷酸化活性，骨髓来源的肥大细胞共培养测定表明颗粒细胞产生功能性KL。在胶原凝胶中培养的生长卵泡中加入10 ng/ml KL导致卵母细胞生长速率增加67%，并且在约50 ng/ml时实现了速率的加倍。抗c-kit单克隆抗体ACK 2严重抑制与卵巢细胞共培养的晚期胎儿和新生卵母细胞的生长，对10- 11日龄小鼠卵泡中培养的卵母细胞生长影响较小。染料木黄酮是一种酪氨酸激酶（包括c-kit）抑制剂，可阻断卵母细胞生长并破坏卵泡形态。在对颗粒细胞中KL产生的调节的初步研究中，我们发现，通过北方分析评估，二丁酰环AMP和生长的卵母细胞都能够诱导颗粒细胞单层中KL mRNA积累增加。这些研究表明，c-kit和KL是维持卵母细胞体外生长所必需的。
Both genetic and descriptive studies have implicated the c-kitreceptor and its ligand, KL, in the process of oocyte growth in the postnatal mouse ovary. In order to test the hypothesis that KL is an oocyte growth factor, we used an oocyte culture system to study its effectsin vitro. Initial experiments established that both ovarian c-kitand KL are biologically active. An immune complex kinase assay demonstrated that ovarian c-kit, found primarily on oocytes, has autophosphorylation activity, and a bone marrow-derived mast cell coculture assay indicated that granulosa cells produce functional KL. The addition of 10 ng/ml KL to growing follicles cultured in collagen gels resulted in a 67% increase in the rate of oocyte growth, and a doubling of the rate was achieved at around 50 ng/ml. ACK2, a monoclonal antibody against c-kit, severely inhibited the growth of late fetal and neonatal oocytes in coculture with ovarian cells and had less effect on growing oocytes cultured in follicles from 10- to 11-day-old mice. Genistein, an inhibitor of tyrosine kinases, including c-kit, blocked oocyte growth and disrupted follicle morphology. In initial studies on the regulation of KL production in granulosa cells, we found that both dibutyryl cyclic AMP and growing oocytes were able to induce increased KL mRNA accumulation in granulosa cell monolayers as assessed by Northern analysis. These studies demonstrate that c-kitand KL are required for maintenance of oocyte growthin vitro.