Methods for quantifying follicular numbers within the mouse ovary
Methods for quantifying follicular numbers within the mouse ovary
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DOI：
10.1530/rep.1.00095
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发表时间：
2004-05-01
期刊：
影响因子：
3.8
通讯作者：
Kerr, JB
中科院分区：
文献类型：
--
作者：
Myers, M;Britt, KL;Kerr, JB
准确估计不同发育阶段的卵泡数量是卵泡发生过程的重要指标，该过程与控制卵母细胞及其支持卵泡细胞生长和成熟的内分泌信号和旁分泌/自分泌机制有关。在使用各种方法的早期研究中报告的相似年龄和/或品系的每个卵巢的卵泡数量存在10倍或更大的差异，导致难以解释对照与实验条件下的卵巢功能。本研究描述了无偏的，无干扰的体视学方法，用于定量小鼠卵巢中早期和生长卵泡的数量。使用分级器方法对成年野生型卵巢的组织切片的限定部分进行采样。原始卵泡和初级卵泡分别用光学和物理解剖法计数。与卵泡大小或形状无关的分离器/剥离器方法估计为1930 +/- 286（S.E.M.）70日龄和100日龄每个卵巢分别有2227 ± 101个原始卵泡和137 ± 25个和265 ± 32个初级卵泡。根据连续切片的精确计数，100日龄时次级和晚期卵泡数估计为每个卵巢135个。使用分离器/物理分离器方法估计了透明带（先前卵泡闭锁的标志物）的残留量，每个卵巢约为500个。所描述的定量方法的应用将有助于更好地理解卵泡动力学和介导其生长和成熟的因素，并允许不同研究之间进行更好的比较。
Accurate estimation of the number of ovarian follicles at various stages of development is an important indicator of the process of folliculogenesis in relation to the endocrine signals and paracrine/autocrine mechanisms that control the growth and maturation of the oocytes and their supporting follicular cells. There are 10-fold or greater differences in follicular numbers per ovary at similar ages and/or strains reported in earlier studies using various methods, leading to difficulties with interpretation of ovarian function in control vs experimental conditions. This study describes unbiased, assumption-free stereological methods for quantification of early and growing follicular numbers in the mouse ovary. A fractionator approach was used to sample a defined fraction of histological sections of adult wild-type ovaries. Primordial and primary follicles were counted independently with the optical and physical disector methods. The fractionator/disector methods, which are independent of follicular size or shape, gave estimations of 1930 +/- 286 (S.E.M.) and 2227 +/- 101 primordial follicles, and 137 +/- 25 and 265 +/- 32 primary follicles per ovary at 70 and 100 days of age respectively. From exact counts on serial sections, secondary and later follicular numbers at 100 days of age were estimated at 135 per ovary. Remnants of zona pellucidae (a marker of previous follicular atresia) were estimated using a fractionator/physical disector approach and were approximately 500 per ovary. The application of the quantitative methods described will facilitate an improved understanding of follicular dynamics and the factors that mediate their growth and maturation and allow for a better comparison between different studies.