Analysis of Differentially Expressed Genes in Ovaries of Polytocous versus Monotocous Dairy Goats Using Suppressive Subtractive Hybridization

Abstract

In this study, suppressive subtractive hybridization (SSH) was performed to screen differentially expressed genes in ovarian tissues between polytocous and monotocous goats. From the SSH cDNA library, we obtained 29 differentially expressed sequence tags (ESTs) that have high similarity with known genes in the public database, which were involved in signal transduction, transcriptional regulation, cellular molecular dynamics, cytoskeleton, metabolism and oxidation-reduction. In addition, one novel EST that has no similarity with known genes in the public database was obtained. Eight ESTs, TIMP1, NUCB1, OAZ1, CXXC5, CAPNS2, ATP5A1, Z and RPL5, were further examined for the reproducibility of the SSH data by the real-time quantitative PCR. The results confirmed an increased expression of respective mRNA in ovarian tissues of polytocous goats compared with those of monotocous goats. The study has identified several genes (known or unknown) that may have effect on follicular development, ovulation and egg activation in goats.

Molecular characterization and hormonal regulation of tissue inhibitor of metalloproteinase 1 in goat ovarian granulosa cells

“…In a previous study, we have demonstrated that Timp1, a differentially expressed gene, was screened from ovarian tissues between polytocous and monotocous Guanzhong dairy goats by suppressive subtractive hybridization [23]. The results confirmed that the different levels of Timp1 mRNA expression in the ovarian tissues of polytocous and monotocous goats could lead to different ovulation rates, which causes a difference in litter size.…”

Identification and verification of differentially expressed genes in the caprine hypothalamic‐pituitary‐gonadal axis that are associated with litter size

“…These results indicate that litter size is a complicated trait. The number of differentially upregulated genes identified herein (>1000) was substantially more than previous reports from An et al (2012) (29 differentially up-regulated genes) or Chen et al (2012) (114 differentially up-regulated genes); this could be due to the tissues chosen for analysis, as the previous work was limited to the ovary whereas we looked at tissue representing the entire HPG axis.…”

“…Three genes identified by our SSH experiment had been previously identified by An et al (2012): ornithine decarboxylase antizyme 1 (OAZ1); ATP synthase, Hþ transporting, mitochondrial F1 complex, alpha subunit 1 (ATP5A1); and ribosomal protein L5 (RPL5). OAZ1 was identified in the hypothalamus and ovary (1 time each) of sexually mature Jining Grey goats.…”

“…The PCR‐based suppressive subtractive hybridization (SSH) technique is used to identify differentially expressed genes (Smits et al, ). The sensitivity of the SSH technique allowed for the identification of genes expressed in sheep ovaries during the reproductive off‐season (Chen et al, ) and in ovaries of polytocous versus monotocous dairy goats (An et al, ). Screening for functional genes of the HPG axis that affect reproduction using molecular biology techniques may facilitate the identification of pathways outside the ovary that are responsible for fertility in goats.…”

OAZ1 knockdown enhances viability and inhibits ER and LHR transcriptions of granulosa cells in geese

“…Our previous study suggested that increased OAZ1 expression might disrupt polyamine homeostasis by inhibiting ODC activity and suppress follicular development in geese [21]. Recently, studies from our and others laboratories indicate that OAZ1 regulates the ovarian and follicular development and ovulation by mediating intracellular polyamine homeostasis in the ovary [21–23]. Granulosa cells play a critical role in ovarian functions such as follicular development, ovulation, and steroidogenesis in both mammals and birds.…”