본 연구는 한우의 자궁내막염에서 발현 변화를 보이는 유전자를 마이크로어레이를 이용하여 조사하였다. 정상적인 자궁내막과 자궁내막염이 있는 자궁내막을 비교한 결과, 전체 확인된 4,560개의 유전자 중 2,026개의 유전자가 자궁내막염에서 증가하였고, 2,534개의 유전자가 감소하였다. 본 연구에서는 상위 조절되는 유전자 10개씩을 정리하였다. 자궁내막염에서 filamin A, pancreatic anionic trypsinogen, Rho GDP dissociation inhibitor alpha, collagen type VI alpha 1, butyrate response factor 2, aggrecanses-2, annexin 14, aminopeptidease A, orphan transporter v7-3 및 epithelial stromal interaction 1의 발현율이 26배 이상 증가하였다. MHC class II antigen, integrin-binding sialoprotein, uterine milk protein precursor, down-regulated in colon cancer 1, glycoprotein 330, dickkopf-1, cfh protein, Ca2+-dependent secretion activator, UL16 binding protein 3 및 proenkephalin은 25.5배 이상 감소하였다. 본 연구에서 얻어진 유전자 정보는 한우의 자궁내막염 진단에 필요한 유용한 생물지표로 사용되어질 수 있을 것으로 사료된다.

Aging causes thymus involution, and genes in thymus play an important role in the development of the immune system. In this study, we compared genes expressed in thymus of neonatal and peripubertal rats using annealing control primers (ACPs)-based GeneFishing polymerase chain reaction (PCR) and semiquantitative reverse transcription (RT)-PCR. We identified 10 differentially expressed genes (DEGs) with 20 ACPs. Of 10 DEGs, bystin-like, collagen type V alpha 1 (COL5A1), and T-cell receptor beta-chain segment 2 (TCRB2) that are related to immune-function were detected in rat thymus. Bystin-like and TCRB2 were up-regulated, while COL5A1 was down-regulated in peripubertal thymus. Semiquantitative RT-PCR confirmed postnatal changes in expression of bystin-like, COL5A1, and TCRB2. These results suggest that bystin-like, COL5A1, and TCRB2 could regulate immune function controlled in thymus as age increases.

Throughout their meiotic maturation in most mammals, oocytes are arrested twice, prophase I and metaphase II. Being released from these arrests, transient or oscillation of intracellular Ca2+ concentration is observed in the ooplasm, which is not answered in relation to the specific role in the resumption of meiotic arrest. Recently, Ca2+/calmodulin-dependent protein kinase II (CaM KII) has been known as a Ca2+ oscillation decoder from the in vitro experiment. CaM KII is multifunctional serine/threonine kinase observed in most cells. Present studies were performed to investigate the role of CaM KII during resumption of meiotic arrest and activation in vitro of mouse oocytes. It was questioned whether CaM KII might be involved in the meiotic resumption of mouse oocytes. Compared to the control, both of CaM KII inhibitors, KN-93 and KN-62, significantly inhibited germinal vesicle breakdown (GVBD) of mouse oocytes in a dose-dependent manner. As the concentration of KN-93 increased, concomitant decrease of intracellular Ca2+ concentration ([Ca2+]i) was also observed using confocal laser scanning microscope (CLSM) and an intracellular Ca2+ indicator, fluo 3-AM. When GVBD oocytes were treated with 6% ethanol, small [Ca2+]i transient was observed in oocytes bathed with Ca2+-free medium and large increase was observed in oocytes bathed with Ca2+-containing medium, suggesting that [Ca2+]i transient could happen from intracellular Ca2+ store as well as Ca2+ influx through Ca2+-channel on the oolemma. However, KN-93 inhibited the [Ca2+]i transient of GVBD oocytes in both cases. Using monoclonal antibodies against α-subunit of CaM KII, tubulin and microtubule-assocaited proteins (MAPs), CaM KII has been colocalized on the spindle with tubulin and MAPs. The present study also demonstrated the presence of α-subunit of CaM KII in heart, kidney, testes, ovary as well as in brain of the mouse. In ovarian follicles, CaM KII was expressed in granulosa cells and oocytes. Based on overall the above results, followings are suggested. First, CaM KII might be involved in the regulatory mechanism of meiotic resumption. Second, CaM KII might play a regulatory role in the stabilization of microtubule.

Sperm specific non selective cation (CatSper) channels belong to the CatSper family of genes and are expressed only in sperm and testis. In general, gene expression profiles in the brains of humans and mice share the highest similarity with those in testis. Therefore, to identify whether CatSper genes are expressed in the mouse brain, we performed reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting. RT-PCR detected all four CatSper mRNAs in both the testis and the brain, with CatSper3 being the most highly expressed. Consistent with RT-PCR data, Western blot analysis showed that CatSper3 was expressed in the brain. We cloned CatSper3 variant 2 from eight-week mouse brain. We named the gene as CatSper3 variant 1 (V1) because the mouse CatSper3 is orthologous to human CatSper3, and another mouse CatSper3 variant (variant 2, V2) with truncated second transmembrane helix was identified. The open reading frame of mCatSper3 V1 consists of 1185 nucleotides and encodes a putative 395-amino acid protein. At the amino acid level, CatSper3 isolated from brain is 100 and 64.8% identical to that isolated from mouse testis and human CatSper3, respectively. Based on comparison between the mCatSper3 V1 ORF and mCatSper3 V2 using TopPred software, the alignment of amino acid sequences shows that the differences exist mainly in segment 2. The CatSper3 transcript consists of eight exons and seven introns, and alternative splice is present within the third exon. In HT22 cell, a mouse hippocampal neuronal cell line, H2O2-induced changes in CatSper3 expression were studied. H2O2 dramatically increased CatSper3 expression in HT22 cells in a dose-and time-dependent manner. The H2O2-induced increase in CatSper3 expression was offset by the addition of N-acetylcysteine (NAC), which is an antioxidant. Taken together, these data strongly indicate that CatSper3 is expressed in mouse brain as variant 1 and suggest that CatSper3 could be a potential target for the modulation of ROS.