Quantification of PERF 15 mRNA in tissue sections from rat testes
Quantification of PERF 15 mRNA in tissue sections from rat testes
复制标题
DOI：
10.1267/ahc.06016
复制
发表时间：
2006-01-01
影响因子：
2.4
通讯作者：
Sasaki, Junzo
中科院分区：
文献类型：
--
作者：
Kogami, Takashi;Miki, Yukari;Sasaki, Junzo
我们以前进行了基础研究，以量化原位杂交（ISH）信号在大鼠睾丸。在该实验模型中，我们选择核糖体RNA（rRNA）作为石蜡切片中的可杂交RNA，因为它允许我们通过图像的“后处理”来容易地定量分析用地高辛（DIG）标记的探针表达的ISH信号。我们应用这种方法来分析转录本PERF 15 mRNA的定量。PERF 15在睾丸中特异性表达，定位于精子头部的刚性细胞骨架结构中，并且被认为参与生精细胞的凋亡过程。信号的量化可有助于阐明PERF 15的详细功能。我们进一步用共聚焦激光扫描显微镜分析了伴随的信号。PERF 15 mRNA在双线期精母细胞中表达最高，在粗线期晚期、双线期精母细胞和早期精子细胞中表达量最高，粗线期早期精母细胞次之，晚期精子细胞最低。PERF 15可能参与导致减数分裂的事件，其中还涉及细胞凋亡。本研究可能有助于确定组织切片中mRNA的浓度。
We previously conducted basic research to quantify in situ hybridization ( ISH) signals in rat testes. In this experimental model, we selected ribosomal RNA ( rRNA) as the hybridizable RNA in paraffin sections, since it allowed us to easily analyze ISH signals expressed with digoxygenin (DIG)-labeled probes quantitatively through "posterization" of the images. We applied this method to analyze the quantification of transcript, PERF 15 mRNA. PERF 15 is expressed specifically in the testes and localized in the rigid cytoskeletal structure of the sperm head, and has been considered to be involved in the apoptotic process of spermatogenic cells. Quantification of the signals may help to clarify the detailed function of PERF 15. We further analyzed the signals concomitant with a confocal laser scanning microscope. The peak of PERF 15 mRNA expression was found in diplotene spermatocytes, and the amount of PERF 15 mRNA was greatest in late pachytene and diplotene spermatocytes and early spermatids, followed by early pachytene spermatocytes, and then late spermatids. PERF 15 may be involved in the events leading to meiotic division, in which apoptosis is also involved. The present study may help to determine the concentration of mRNA in tissue sections.