Reproductive failure in mice lacking inter-alpha-trypsin inhibitor (ITI) – ITI target genes in mouse ovary identified by microarray analysis

Abstract

Bikunin, a Kunitz-type protease inhibitor, is found in blood and urine. It has been established by two laboratories independently that the bikunin knockout female mice display a severe reduction in fertility: the cumulus oophorus has a defect in forming the extracellular hyaluronanrich matrix during expansion. Proteins of the inter-alphatrypsin inhibitor (ITI) family are eliminated in mice in which the bikunin gene has been inactivated, since bikunin is essential for their biosynthesis. Proteins of the ITI family may contribute to the microenvironment in which ovulation takes place. It is not clear, however, whether a single mechanism affects the reproductive function including ovulation. For identifying the full repertoire of the ITI deficiency-related genes, a cDNA microarray hybridization screening was conducted using mRNA from ovaries of wild-type or bik / female mice. A number of genes were identified and their regulation was confirmed by real-time RT-PCR analysis. Our screen identified that 29 (0·7%) and 5 genes (0·1%) of the genes assayed were, respectively, up-and down-regulated twofold or more. The identified genes can be classified into distinct subsets. These include stress-related, apoptosis-related, proteases, signaling molecules, agingrelated, cytokines, hyaluronan metabolism and signaling, reactive oxygen species-related, and retinoid metabolism, which have previously been implicated in enhancing follicle development and/or ovulation. Real-time RT-PCR analysis confirmed that these genes were up-and down-regulated two-to tenfold by bikunin knockout. These studies demonstrate that proteins of the ITI family may exert potent regulatory effects on a major physiological reproductive process, ovulation.

Hormone-sensitive lipase deficiency alters gene expression and cholesterol content of mouse testis

“…The transcriptome data identified the genes involved in testis-specific biological processes (Musiolik 1971, Lee et al . 2012, Waclawska & Kurpisz 2012). HSL knockout affected the expression levels of many genes of lipid metabolism because HSL hydrolyzes glycerol and cholesterol esters.…”

Extracellular matrix in ovarian follicular development and disease

“…Bikunin and other factors cDNA microarray screening of ovaries from wild-type and bikunin null mice have identified the up-regulation of Cdc-42, which may prevent COC expansion by the up-regulation of cadherins in the pre-ovulatory follicle (Suzuki et al 2004). Cathepsin L, which is induced in pre-ovulatory follicles by the LH surge (Robker et al 2000), is also up-regulated in Bik -/-mice, as is the anti-apoptotic Bcl-2 protein.…”

“…Cathepsin L, which is induced in pre-ovulatory follicles by the LH surge (Robker et al 2000), is also up-regulated in Bik -/-mice, as is the anti-apoptotic Bcl-2 protein. Other up-regulated genes that may be important in the reproductive phenotype of bikunin-deficient mice include granulocytecolony stimulating factor, IL-2 and IL-5 receptor and protein kinase C (Suzuki et al 2004). Both cathepsin L and ADAMTS-1 are proteases whose gene expression is regulated by LH and the progesterone receptor (Robker et al Fig.…”

Expression and localization of Inter-alpha Inhibitors in rodent brain

“…Administration of exogenous bikunin and IAIPs has been shown to attenuate systemic inflammation (Lim et al, 2003, Singh et al, 2010) and inhibit tumor metastasis (Kobayashi et al, 2003, Suzuki et al, 2004). IAIPs exert beneficial effects in sepsis and systemic inflammatory disorders both in neonates and in adults (Lim et al, 2003, Singh et al, 2010).…”