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Rhonda D Kineman, Manuel D Gahete and Raul M Luque

variant’. This transcript was found to be dominantly expressed in the mouse hypothalamus (HPT) and pituitary (PIT), but was not detectable in the stomach. Physiologic relevance of In2-ghrelin variant is supported by the fact that mRNA levels are regulated

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Ying Zhang, Mingtong Xu, Shaoling Zhang, Li Yan, Chuan Yang, Wensheng Lu, Yan Li and Hua Cheng

preparation and transfection Three siRNAs were designed from the mouse GPR40 cDNA sequence (EMBL/GenBank/DDBJ accession no. AF539809). These siRNAs, containing 21 nucleotides, were synthesized by Ambion, Austin, TX, USA. The sense sequences targeted

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Luc J Martin and Jacques J Tremblay

identified the orphan nuclear receptor NUR77 (NR4A1) as a rapidly and strongly induced transcription factor that contributes to mouse Star transcription in Leydig cells in response to cAMP/forskolin stimulation ( Martin et al . 2008 ). The orphan nuclear

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Maxime A Tremblay, Raifish E Mendoza-Villarroel, Nicholas M Robert, Francis Bergeron and Jacques J Tremblay

INSL3 promoter and found that this element is recognized and activated by the KLF6 transcription factor, which we have located in the nuclei of somatic cells of the mouse testis. Furthermore, we report that KLF6 functionally cooperates with the nuclear

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Xiaohui Wang, Yidong Li, Xiaoyan Zhu, Yan Wang, Fei Diao and Jian Lu

antiproliferation of GC/GR by examining 1) the effect of GC/GR on the expression of SIRPA and its possible role in the antiproliferative effect of GC/GR in RAW264.7 cells and mouse peritoneal macrophage; 2) changes in the expression of CDK 2, cyclinD1, and cyclinB1

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S Miyagawa, A Suzuki, Y Katsu, M Kobayashi, M Goto, H Handa, H Watanabe and T Iguchi

Developmental exposure to a synthetic estrogen, diethylstilbestrol (DES), induces carcinogenesis in human and laboratory animals. In mice, neonatal DES treatment induces persistent proliferation and keratinization of the vaginal epithelium, even in the absence of the ovaries, resulting in cancerous lesions later in life. To understand the mechanisms underlying this persistent cell proliferation and differentiation, we characterized the gene expression patterns in the neonatally DES-exposed mouse vagina using DNA microarray and real-time quantitative RT-PCR. We found that genes related to cellular signaling, which are candidates for mediating the persistent proliferation and differentiation, were altered, and genes related to the immune system were decreased in the neonatally DES-exposed mouse vagina. We also noted high expression of interleukin-1 (IL-1)-related genes accompanied by phosphorylation of JNK1. In addition, expression IGF-I and its binding proteins was modulated and led to phosphorylation of IGF-I receptor and Akt, which is one of the downstream factors of IGF-I signaling. This led us to characterize the expression as well as the phosphorylation status of IL-1 and IGF-I signaling pathway components which may activate the phosphorylation cascade in the vagina of mice exposed neonatally to DES. These findings give insight into persistent activation in the vagina of mice exposed neonatally to DES.

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B A Evans, M John, K J Fowler, R J Summers, M Cronk, J Shine and G W Tregear

ABSTRACT

Relaxin is a polypeptide hormone that has a variety of physiological effects both on remodelling of collagen and on uterine contractility. These are most apparent during pregnancy. The sequences of relaxin cDNAs derived from ovaries of late-pregnant random-bred Swiss mice have been established. Multiple subclones obtained from three independent polymerase chain reaction experiments were found to encode relaxins which were identical except at position 11 in the A chain (Ile or Val). All mouse relaxin cDNAs expressed in the ovary during pregnancy had an extra tyrosine inserted prior to the final A chain cysteine residue, a result confirmed by direct sequencing of relaxin peptides. Whilst this tyrosine insertion must have local effects on the folding of the A chain, structure—activity studies will clarify whether it perturbs functional interaction with the relaxin receptor. We have shown that there is a single relaxin gene in the mouse genome, and that expression during pregnancy occurs in the ovary but is not detectable in the placenta, uterus or fetus.

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Young Sun Kang, Yun Gyu Park, Bo Kyung Kim, Sang Youb Han, Yi Hwa Jee, Kum Hyun Han, Mi Hwa Lee, Hye Kyoung Song, Dae Ryong Cha, Shin Wook Kang and Dae Suk Han

of fibronectin expression and cellular growth in rat mesangial cells ( Reddy et al. 2002 ). In this study, we examined the effects of Ang-II on VEGF mRNA expression and protein production in cultured mouse podocytes, and evaluated whether

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J. C. Pascall and K. D. Brown

ABSTRACT

The 5′-flanking sequence of the mouse epidermal growth factor gene has been isolated from a mouse genomic DNA library. S1 nuclease mapping indicated that the transcription start sites used in the submaxillary gland and the kidney are identical. Computer-aided sequence comparisons have indicated regions of the gene which may be involved in hormonal regulation.

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NJ Pavlos, J Xu, JM Papadimitriou and MH Zheng

Small GTP-binding proteins of the Rab subfamily are key regulators of intracellular vesicle transport. Here we report the isolation of a cDNA clone encoding the complete Rab3c isoform from mouse embryo using a degenerative PCR-based approach. Multiple sequence alignment revealed that the predicted amino acid sequence was identical to the previously identified rat Rab3c isoform and 98% identical to the published bovine Rab3c GTPase from brain. Furthermore by in situ hybridisation, Rab3c mRNA was detectable within various regions of the brain, cartilage and highly enriched within intestinal villi of foetal tissues. Chondrocytes in the hypertrophic zone, but not reserve or proliferative zones, expressed high levels of Rab3c. This pattern of expression corresponds with the genesis of matrix vesicles during endochondral ossification.In all, our results suggest that in addition to its functional role during regulated secretion in brain, Rab3c may play a part in matrix vesicle trafficking during skeletal development.