Reproductive toxicity has been an exciting topic of research in reproductive biology in recent years. Soluble fluoride salts are toxic at high concentrations; their reproductive toxicity was assessed in this study by administering different fluoride salt concentrations to mice. Continuous feeding for five weeks resulted in damage to the histological architecture of ovaries. The expression of genes, including Dazl, Stra8, Nobox, Sohlh1, and ZP3 gene, associated with oocyte formation were much lower in the experimental group as compared with the control group. The number of in vitro fertilization of mature oocytes were also much lower in the experimental group as compared with control. Moreover, the fertility of female mice, as assessed by mating with normal male mice, was also lower in experimental compared with control groups. The expression of the oocyte-specific genes: Bmp15, Gdf9, H1oo, and ZP2, which are involved in oocyte growth and the induction of the acrosome reaction, decreased with the fluoride administration. DNA methylation and histone acetylation (H3K18ac and H3K9ac) are indispensable for germline development and genomic imprinting in mammals, and fluoride administration resulted in reduced levels of H3K9ac and H3K18ac in the experimental group as compared with the control group, as detected by immunostaining. Our results indicate that the administration of high concentrations of fluoride to female mice significantly reduced the number of mature oocytes and hampered their development and fertilization. Thus, this study lays a foundation for future studies on fluoride-induced reproductive disorders in women.

/pdf/adverse-effects-of-high-concentrations-of-fluoride-on-2qbcieco5p.pdf

Adverse Effects of High Concentrations of Fluoride on Characteristics of the Ovary and Mature Oocyte of Mouse.

세계적으로 양식이 되고 있는 어패류 중 참굴(Crassostrea gigas)은 생산량과 가격에 있어서 가장 중요한 양식 품종의 하나 로 자리 잡고 있다(FAO, 2014). 우리나라에서 굴은 1897년 원산만에서 처음 양식되었으며, 1960년 이후부터 경남지역을 중심으로 연승수하식 방법이 널 리 보급되면서 생산량이 증가하였다. 2013년에는 연간 30,308 톤이 생산되었으며, 이중 경남지역이 25,906톤, 전남지역이 3,992톤을 생산하였다(KSIS, 2014). 우리나라 굴 소비는 산란이 끝난 10월에서 이듬해 2-3월 사 이이며, 이 시기에 굴은 산란에서 회복이 되고 체내에 글리코 겐을 축적하여 상품성이 높아지게 된다(Kim et al., 2009). 생 식소 발달과 더불어 저장되어 있는 에너지를 5-8월까지 소비 하고 가을과 겨울에 에너지를 다시 저장하는 것으로 알려져 있 다(Delaporte et al., 2006; Mallet et al., 2006). 특히 산란시기 는 비만도와 맛이 저하되며, 근육 조직이 다른 패류보다도 연하 여 소화 분해되기 쉬운 특성이 있어 식중독을 일으킬 확률이 높 고, 여름철 미생물 오염도 높아 굴 섭취량이 적은 것으로 사료 된다. 그러나 최근 굴 국밥 등 굴요리 전문점의 성업으로 냉동 굴보다는 생굴을 선호하고 있어 여름철에도 비계통적으로 생산 되고 있는 실정이다. 경남 거제만 및 자란만 굴(Crassostrea gigas)의 일반성분, pH, 글리코겐 함량의 월별 변화

Seasonal Variation in the Proximate Composition, pH and Glycogen Content of Oysters Crassostrea gigas Collected in Geoje and Jaran Bay in Korea

Epigenetic perturbations during the reprogramming process have been described as the primary cause of the low efficiency of somatic cell nuclear transfer (SCNT). In this study, we tested three strategies targeting nuclear reprogramming to investigate effects on equine SCNT. First, we evaluated the effect of treating somatic cells with chetomin, a fungal secondary metabolite reported to inhibit the trimethylation on histone 3 lysine 9 (H3K9 me3). Second, caffeine was added to the culture medium during the enucleation of oocytes and before activation of reconstructed embryos as a protein phosphatase inhibitor to improve nuclear reprogramming. Third, we tested the effects of the histone deacetylase inhibitor trichostatin A (TSA) added during both activation and early embryo culture. Although none of these treatments significantly improved the developmental rates of the invitro aggregated cloned equine embryos, the first equine cloned foal born in Australia was produced with somatic cells treated with chetomin. The present study describes the use of chetomin, caffeine and TSA for the first time in horses, serving as a starting point for the establishment of future protocols to target epigenetic reprogramming for improving the efficiency of equine cloning. Cloning is an expensive and inefficient process, but has gained particular interest in the equine industry. In this study we explored different strategies to improve cloning efficiency and produced the first cloned foal born in Australia. Our data serve as a starting point for the establishment of future protocols for improving equine cloning efficiency.

Targeting epigenetic nuclear reprogramming in aggregated cloned equine embryos

Levels of maturation-promoting factor (MPF) in oocytes decline after vitrification, and this decline has been suggested as one of the main causes of low developmental competence resulting from cryoinjury. Here, we evaluated MPF activity in vitrified mouse eggs following treatment with caffeine, a known stimulator of MPF activity, and/or the proteasome inhibitor MG132. Collected MII oocytes were vitrified and divided into four groups: untreated, 10 mM caffeine (CA), 10 μM MG132 (MG), and 10 mM caffeine +10 μM MG132 (CA+MG). After warming, the MPF activity of oocytes and their blastocyst formation and implantation rates in the CA, MG, and CA+MG groups were much higher than those in the untreated group. However, the cell numbers in blastocysts did not differ among groups. Analysis of the effectiveness of caffeine and MG132 for improving somatic cell nuclear transfer (SCNT) technology using cryopreserved eggs showed that supplementation did not improve the blastocyst formation rate of cloned mouse eggs. These results suggest that maintaining MPF activity after cryopreservation may have a positive effect on further embryonic development, but is unable to fully overcome cryoinjury. Thus, intrinsic factors governing the developmental potential that diminish during oocyte cryopreservation should be explored.

Maintained MPF Level after Oocyte Vitrification Improves Embryonic Development after IVF, but not after Somatic Cell Nuclear Transfer.

Implantable devices comprising a scaffold, cells, and/or therapeutic molecules are provided. Uses include inhibition of pathological immunity, elaboration of therapeutic growth factors, and immune augmentation. The implantable device may contain a porous substrate structure to which therapeutic cells are adherent, which can be implanted and explanted from a patient. In another embodiment, a medical device includes a porous substrate seeded with therapeutic cells, said device comprising of an additional layer allowing free exchange of molecules without cell escape. Said medical device is seeded with thymic medullary epithelial cells or progenitors of said thymic medullary epithelial cells. In conditions where tolerance to alloreactive donor cells is desired, the thymic medullary epithelial cells or progenitors thereof are derived from recipient cells. In conditions where tolerance to alloreactive recipient cells is desired, the thymic medullary epithelial cells or progenitors thereof are derived from donor cells.

Implantable cellular therapy device for treatment of graft versus host disease and tolerance induction

The development of embryos reconstructed by nuclear transfer (NT) is dependent upon numerous factors. Central to development is the quality and development of the recipient cytoplast. Typically oocytes Corresponding author: Joon-Hee Lee Tel: +82-55-772-1886 Fax: +82-55-772-1889 E-mail: sbxjh@gnu.ac.kr 168 ... Journal of Agriculture & Life Science 47(6) at either AI/TI or MII are potential candidates used as recipient cytoplasts for NT, because they contain active MPF, which causes NEBD and PCC in the transferred nucleus and may be essential for nuclear reprogramming. In vitro maturation of porcine oocytes generally spend longer times (44-48h), progress asynchronously from immature stage and also present insufficient MPF for NEBD and PCC. Here, we have developed three stages establishment of in vitro porcine oocyte maturation to select good quality of oocytes used as recipient cytoplasts for NT. Firstly, oocytes were assessed by the activity of glucose-6-phosphate dehydrogenase (G6PD) before in vitro oocyte maturation. Positively BCB stained oocytes reached to MII (67.0% vs. 58.4%) and produced more parthenogenetic blastocysts (50.2% vs. 29.6%) as compared with negatively BCB stained oocytes (P < 0.05). Secondly, positively BCB stained oocytes were treated with 5 μg/ml CHXM to block meiotic progression for synchronizing the cell cycle of oocytes for 16 hr. All of oocytes (130/130) were efficiently synchronized at GV stage by treatment with CHXM, whereas most of oocytes (w/o CHXM) were passed through GV stage (80/176). In addition, retrieval from treatment of CHXM could reversibly induce meiotic resumption and progresses synchronously in porcine oocytes. However, there was no difference between CHXM treated and non-treated group (21.9% vs. 22.3%) in the developmental rate to the blastocyst stage of parthenogenetic embryos. Following treatment of 5 mM caffeine (a phosphatase inhibitor) for 12 hr, all of oocytes were significantly increased the activity of MPF (P < 0.05). Finally, a piglet was produced from NT embryos reconstructed by using non-caffeine treated recipients.

Selection and Synchronization of recipient cytopalsts for improving the efficiency of porcine somatic cell nuclear transfer

Somatic cell nuclear transfer (SCNT) has generally demonstrated that a differentiated cell can convert into a undifferentiated or pluripotent state. In the SCNT experiment, nuclear reprogramming is induced by exposure of introduced donor nuclei to the recipient cytoplasm of matured oocytes. However, because the efficiency of SCNT still remains low, a combination of SCNT technique with the ex-ovo method may improve the normal development of SCNT embryos. Here we hypothesized that treatment of somatic cells with extracts prepared from the germinal vesicle (GV) stage Siberian sturgeon oocytes prior to their use as nuclear donor for SCNT would improve in vitro development. A reversible permeability protocol with 4 μg/mL of digitonin for 2 min at 4°C in order to deliver Siberian sturgeon oocyte extract (SOE) to porcine fetal fibroblasts (PFFs) was carried out. As results, the intensity of H3K9ac staining in PFFs following treatment of SOE for 7 h at 18°C was significantly increased but the intensity of H3K9me3 staining in PFFs was significantly decreased as compared with the control (p<0.05). Additionally, the level of histone acetylation in SCNT embryos at the zygote stage was significantly increased when reconstructed using SOE-treated cells (p<0.05), similar to that of IVF embryos at the zygote stage. The number of apoptotic cells was significantly decreased and pluripotency markers (Nanog, Oct4 and Sox2) were highly expressed in the blastocyst stage of SCNT embryos reconstructed using SOE-treated cells as nuclear donor (p<0.05). And there was observed a better development to the blastocyst stage in the SOE-treated group (p<0.05). Our results suggested that pre-treatment of cells with SOE could improve epigenetic reprogramming and the quality of porcine SCNT embryos.

/pdf/siberian-sturgeon-oocyte-extract-induces-epigenetic-1iv0nqr0tj.pdf

Siberian Sturgeon Oocyte Extract Induces Epigenetic Modifications of Porcine Somatic Cells and Improves Developmental Competence of SCNT Embryos

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Selection and Synchronization of recipient cytopalsts for improving the efficiency of porcine somatic cell nuclear transfer

Siberian Sturgeon Oocyte Extract Induces Epigenetic Modifications of Porcine Somatic Cells and Improves Developmental Competence of SCNT Embryos