Effects of small-molecule compounds on fibroblast properties in golden snub-nosed monkey (Rhinopithecus roxellana).

BACKGROUND: Golden snub-nosed monkey (Rhinopithecus roxellana) is an endangered primate species, whose molecular material for conservation purposes has not yet been maintained. Although small-molecule compounds (SMCs) have been reported to improve induced pluripotent stem cells (iPSCs), their efficiency in the interspecies-transferred nucleus is still unknown. METHODS: We thus used the fibroblasts from the golden snub-nosed monkey treated with SMC as donor cells, injected into the enucleated oocytes of goats, to test such efficiency. Gene expression profiles in the cell-constructed embryos with and without SMCs were compared by qPCR. RESULTS: The results show that cell morphology undergoes remarkable changes (volume is smaller than normal cells, and many black spots in the cytoplasm were found); pluripotent genes (Oct4, Sox2, and Nanog) significantly increased with SMC treatment. CONCLUSIONS: This study demonstrates that SMCs alter the properties of donor cells and promote the expression of pluripotent genes in hybrid embryos.

Effects of Alcohol on Mitochondrial Functions of Cumulus Cells in Mice.

Alcohol is an important compound used in food, agriculture, and medicine. In this study, we investigated the effect of alcohol on oocyte quality in mice by exposing animals for different duration times during an estrous cycle. Cumulus-oocyte complexes were collected from mice after pregnant mare serum gonadotropin- and human chorionic gonadotropin-induced superovulation. Ovulation number, E2 level in serum, and parthenogenetic embryo development in vitro were evaluated. Mitochondrial gene expression, mitochondrial membrane potential, and reactive oxygen species (ROS) levels in the cumulus were also assessed. The results showed that acute exposure to alcohol did not affect ovulation time (p > 0.05). Blasocyst formation rate in vitro was significantly improved after 1 and 2 days of alcohol exposure (p < 0.01). Mitochondrial membrane potential was significantly increased after 1-4 days of alcohol exposure (p < 0.05), but it decreased after 5 days (p < 0.05). ROS levels remained relatively low after 2, 3, and 4 days of exposure (p < 0.05), and they significantly increased after 6 days (p < 0.05). In addition, alcohol altered the expression of mitochondrial and nuclear genes in the cumulus. Taken together, our data suggest that acute exposure to alcohol affects oocyte quality by influencing the function and gene expression in the cumulus. These results underscore potential implications for the development of human reproductive therapeutics.

A novel tissue inhibitor of metalloproteinase in blood clam Tegillarca granosa: molecular cloning, tissue distribution and expression analysis.

Tissue inhibitor of metalloproteinases (TIMPs) were originally characterized as inhibitors of matrix metalloproteinases (MMPs), but their range of activities has been found to be broader as it includes the inhibition of several of the MMPs, etc. The cDNA encoding TIMP-4-like gene from blood clam Tegillarca granosa (designated as Tg-TIMP-4-like) which is the first tissue inhibitor of metalloproteinase identified in blood clams, was cloned and characterized. It was of 1164 bp, and an open reading frame (ORF) of 666 bp encoding a putative protein of 222 amino acids. The predicted amino acid sequence comprised all recognized functional domains found in other TIMP homologues and showed the highest (30.56%) identity to the TIMP-1.3 from Crassostrea gigas. Several highly conserved motifs including several TIMP signatures, amino acid residue Cys³° responsible for coordinating the metal ions, the Cys-X-Cys motif and the putative NTR (netrin) domain were almost completely conserved in the deduced amino acid of Tg-TIMP-4 like, which indicated that Tg-TIMP-4-like should be a member of the TIMP family. The mRNA expression of Tg-TIMP-4-like in the tissues of mantle, adductor muscle, foot, gill, hemocyte and hepatopancreas was examined by quantitative real-time PCR (qT-PCR) and mRNA transcripts of Tg-TIMP-4-like were mainly detected in hemocyte, and weakly detected in the other tissues. We also observed that Tg-TIMP-4 like mRNA accumulated significantly during Vibrio parahaemolyticus, Peptidogylcan (PGN) and Lipopolysaccharide (LPS) challenge, whereas the timing and quantitative differences of mRNA expression against different challenge indicated that Tg-TIMP-4-like may play a pivotal role in mollusc defense mechanisms.