Improvement in identification of pro-estrous mice by using a novel method of detecting vaginal mucous cells.
Improvement in identification of pro-estrous mice by using a novel method of detecting vaginal mucous cells.
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通过使用检测阴道粘液细胞的新方法改进发情前小鼠的识别。
DOI：
10.1007/s00441-020-03310-w
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发表时间：
2021
影响因子：
3.6
通讯作者：
Kazuki Yoshioka
中科院分区：
文献类型：
--
作者：
Makoto Sugiyama;Arata Yasunaga;Ryosuke Kobayashi;Hanae Fukasawa;Osamu Hashimoto;Shiro Kurusu;Hiroshi Sasada;Kazuki Yoshioka
使用阴道脱落细胞学准确鉴定小鼠发情周期是该物种受控繁殖的初始且关键的步骤。然而，通常很难区分周期的每个阶段，从而选择发情前的小鼠进行交配。为了提高发情前阶段识别的准确性，我们重新评估阴道皱襞组织学并修改脱落细胞学方法。使用卡诺溶液中的甲醇而不是多聚甲醛进行组织固定，结合阿尔新蓝染色而不是传统的吉姆萨染色，可以更好地显示角化层上方阴道上皮中的粘膜细胞层。发现皱襞组织学中的这种粘液层专门在发情前和动情晚期阶段形成，并且涂片样本中的粘液细胞仅在发情前阶段脱落。通过盲法测试发现，与传统方法相比，这种新方法可以提高发情前阶段的准确识别率（80% vs 50%）。与这一发现一致的是，通过新方法选择的“发情前”雌性的交配实验显示，成功率显着高于传统方法（78.0% vs 47.5%）。因此，我们的研究表明，阴道脱落粘液细胞是检测雌性小鼠“接受”状态的更好的潜在标记，从而提高交配成功率。
Accurate identification of the murine estrous cycle using vaginal exfoliative cytology is the initial and crucial step for controlled reproduction of this species. However, it is generally difficult to discriminate each stage of the cycle, and thus to select pro-estrous mice for mating. To increase the accuracy of identification of the pro-estrous stage, we re-evaluated the vaginal fold histology and modified the method of exfoliative cytology. Tissue fixation using methanol in Carnoy’s solution but not paraformaldehyde, combined with Alcian blue staining but not the conventional Giemsa staining, resulted in better manifestation of mucosal cell layers in the vaginal epithelium just above the keratinized layer. This mucous layer in the fold histology was found to form specifically in the pro-estrous and late di-estrous stages, and the mucous cells exfoliated in smear samples only in the pro-estrous stage. This novel method was found, by a blinded test, to increase the rate of accurate identification of the pro-estrous stage compared to the conventional method (80% vs 50%). Consistent with this finding, the mating experiment with “pro-estrous” females selected by the novel method revealed a significantly higher success rate than that with the conventional method (78.0% vs 47.5%). Thus, our study demonstrates vaginal exfoliative mucous cells as a better potential marker to detect the “receptive” state of female mice that leads to an improved success rate of mating.