Quantification of in situ hybridization signals in rat testes

被引：8

作者：

Kimura, T ^{[1
]}

Kosaka, J ^{[1
]}

Nomura, T ^{[1
]}

Yamada, T ^{[1
]}

Miki, Y ^{[1
]}

Takagi, K ^{[1
]}

Kogami, T ^{[1
]}

Sasaki, J ^{[1
]}

机构：

[1] Okayama Univ, Grad Sch Med & Dent, Dept Cytol & Histol, Okayama 7008558, Japan

来源：

JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY | 2004年 / 52卷 / 06期

关键词：

quantification; posterization; in situ hybridization; rat; testis; ribosomal RNA;

D O I：

10.1369/jhc.4A6249.2004

中图分类号：

Q2 [细胞生物学];

学科分类号：

071009 [细胞生物学]; 090102 [作物遗传育种];

摘要：

We performed basic research into quantifying in situ hybridization (ISH) signals in rat testis, a suitable organ for the quantification because germ cells undergo synchronized development and show stage-specific gene expression. In this model experiment, rRNA was selected as the hyloridizable RNA in paraffin sections. Specimens fixed with Bouin's fixative and hybridized with digoxygenin-labeled probes could easily be analyzed quantitatively through "posterization" of the images. The amount of rRNA hybridized with the probe was greatest in early primary spermatocytes, followed by pachytene primary spermatocytes, then diplotene spermatocytes, and finally by secondary spermatocytes and spermatids. The amounts reached low levels in metaphase, anaphase, and telophase of meiotic division and early step 1 spermatids, and then slightly increased during spermiogenesis. ISH rRNA staining was a useful parameter for evaluation of the quantitative analysis of mRNA and the levels of hybridizable RNA in tissue sections.

引用

页码：813 / 820

页数：8

共 24 条

[1]

FUNDAMENTALS OF QUANTITATIVE AUTORADIOGRAPHY BY COMPUTER DENSITOMETRY FOR IN-SITU HYBRIDIZATION, WITH EMPHASIS ON P-33 [J].