Gene expression analysis of the pro-oestrous-stage rat uterus reveals neuroligin 2 as a novel steroid-regulated gene
Han-Seung Kang* A , Chae-Kwan Lee* B , Ju-Ran Kim C , Seong-Jin Yu C , Sung-Goo Kang C E , Deog-Hwan Moon B , Chang-Hee Lee B and Dong-Kyoo Kim D
+ Author Affiliations
- Author Affiliations
A Department of Life Science, College of Natural Sciences, Hanyang University, Seoul 133-791, South Korea.
B Institute of Industrial Medicine and Department of Occupational and Environmental Medicine, Paik Hospital, Inje University, Busan 614-735, South Korea.
C School of Biotechnology and Biomedical Science, Institute of Basic Sciences, Inje University, Kimhae 621-749, South Korea.
D Department of Chemistry, College of Natural Sciences, Inje University, Kimhae 621-749, South Korea.
E Corresponding author. Email: biosgkan@inje.ac.kr
Reproduction, Fertility and Development 16(8) 763-772 https://doi.org/10.1071/RD04040
Submitted: 29 March 2004 Accepted: 7 November 2004 Published: 13 January 2005
Abstract
In the present study, differential gene expression in the uteri of ovariectomised (OVX) and pro-oestrous rats (OVX v. pro-oestrus pair) was investigated using cDNA expression array analysis. Differential uterine gene expression in OVX rats and progesterone (P4)-injected OVX rats (OVX v. OVX + P4 pair) was also examined. The uterine gene expression profiles of these two sets of animals were also compared for the effects of P4 treatment. RNA samples were extracted from uterine tissues and reverse transcribed in the presence of [α32P]-dATP. Membrane sets of rat arrays were hybridised with cDNA probe sets. Northern blot analysis was used to validate the relative gene expression patterns obtained from the cDNA array. Of the 1176 cDNAs examined, 23 genes showed significant (>two-fold) changes in expression in the OVX v. pro-oestrus pair. Twenty of these genes were upregulated during pro-oestrus compared with their expression in the OVX rat uterus. In the OVX v. OVX + P4 pair, 22 genes showed significant (>two-fold) changes in gene expression. Twenty of these genes were upregulated in the OVX + P4 animals. The genes for nuclear factor I–XI, afadin, neuroligin 2, semaphorin Z, calpain 4, cyclase-associated protein homologue, thymosin β-4X and p8 were significantly upregulated in the uteri of the pro-oestrus and OVX + P4 rats of both experimental pairs compared with the OVX rat uteri. These genes appear to be under the control of P4. One of the most interesting findings of the present study is the unexpected and marked expression of the neuroligin 2 gene in the rat uterus. This gene is expressed at high levels in the central nervous system and acts as a nerve cell adhesion factor. According to Northern blot analysis, neuroligin 2 gene expression was higher during the pro-oestrus and metoestrus stages than during the oestrus and dioestrus stages of the oestrous cycle. In addition, neuroligin 2 mRNA levels were increased by both 17β-oestradiol (E2) and P4, although P4 administration upregulated gene expression to a greater extent than injection of E2. These results indicate that neuroligin 2 gene expression in the rat uterus is under the control of both E2 and P4, which are secreted periodically during the oestrous cycle.
Extra keywords: neurexin, PDZ domain, post-synaptic scaffolding protein-95, proliferation.
Acknowledgments
This work was supported by grant no. 2000-1-20700-002-3 from the Basic Research Program and BPRC (2004) of the KOSEF, and in part by the Inje Research Foundation and Post-master Programme (2004).
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* H.-S. Kang and C.-K. Lee contributed equally to this paper.
