Oligodendrocytes (OLs) are myelinating glial cells that form myelin sheaths around axons to ensure rapid and focal conduction of action potentials. Here, we found that an axonal outgrowth regulatory molecule, AATYK (apoptosis-associated tyrosine kinase), was up-regulated with OL differentiation and remyelination. We therefore studied its role in OL differentiation. The results showed that AATYK knockdown inhibited OL differentiation and the expression of myelin genes in vitro. Moreover, AATYK-deficiency maintained the proliferation status of OLs but did not affect their survival. Thus, AATYK is essential for the differentiation of OLs.
Animals ; Animals, Newborn ; Apoptosis Regulatory Proteins ; genetics ; metabolism ; Cell Differentiation ; drug effects ; physiology ; Cell Proliferation ; drug effects ; genetics ; Cells, Cultured ; Cuprizone ; toxicity ; Demyelinating Diseases ; chemically induced ; metabolism ; pathology ; Embryo, Mammalian ; Gene Expression Regulation, Developmental ; genetics ; Ki-67 Antigen ; metabolism ; Mice ; Mice, Inbred C57BL ; Myelin Basic Protein ; metabolism ; Myelin Proteolipid Protein ; metabolism ; Myelin Sheath ; drug effects ; metabolism ; Oligodendroglia ; drug effects ; metabolism ; Protein-Tyrosine Kinases ; genetics ; metabolism ; RNA, Small Interfering ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley

As the number of young cancer survivors increases, quality of life after cancer treatment is becoming an ever more important consideration. According to a report from the American Cancer Society, approximately 810,170 women were diagnosed with cancer in 2015 in the United States. Among female cancer survivors, 1 in 250 are of reproductive age. Anticancer therapies can result in infertility or sterility and can have long-term negative effects on bone health, cardiovascular health as a result of reproductive endocrine function. Fertility preservation has been identified by many young patients diagnosed with cancer as second only to survival in terms of importance. The development of fertility preservation technologies aims to help patients diagnosed with cancer to preserve or protect their fertility prior to exposure to chemo- or radiation therapy, thus improving their chances of having a family and enhancing their quality of life as a cancer survivor. Currently, sperm, egg, and embryo banking are standard of care for preserving fertility for reproductive-age cancer patients; ovarian tissue cryopreservation is still considered experimental. Adoption and surrogate may also need to be considered. All patients should receive information about the fertility risks associated with their cancer treatment and the fertility preservation options available in a timely manner, whether or not they decide to ultimately pursue fertility preservation. Because of the ever expanding number of options for treating cancer and preserving fertility, there is now an opportunity to take a precision medicine approach to informing patients about the fertility risks associated with their cancer treatment and the fertility preservation options that are available to them.
Adult Stem Cells ; Cell Culture Techniques ; Cryopreservation/*methods ; *Embryo, Mammalian ; Female ; Fertility Preservation/*methods ; Humans ; Neoplasms/drug therapy/*therapy ; *Oocytes ; Ovarian Follicle/drug effects/metabolism/transplantation ; *Ovary/transplantation ; Ovulation Induction/methods ; Precision Medicine

The tyrosine kinase system angiopoietin (Ang)/Tie interacts with vascular endothelial growth factor pathway and regulates vessel quiescence in adults as well as later steps of the angiogenic cascade related to vessel maturation. Since all Angs are able to bind to Tie-2 but none binds to Tie-1, the function of Tie-2 and its ligands have captured attention. However, emerging evidence indicates unique roles of the orphan receptor Tie-1 in angiogenesis under physiological and pathological conditions. It is required for maintaining vascular endothelial cell integrity and survival during murine embryo development and in adult and may be involved in modulating differentiation of hematopoietic cells in adult. Tie-1 exhibits poor tyrosine kinase activity and signals via forming heterodimers with Tie-2, inhibiting Tie-2 signaling mediated by Angs. This inhibition can be relieved by Tie-1 ectodomain cleavage mediated by tumor- and inflammatory-related factors, which causes destabilization of vessels and initiates vessel remodeling. Up-regulated Tie-1 expression has been found not only in some leukemia cells and tumor related endothelial cells but also in cytoplasm of carcinoma cells of a variety of human solid tumors, which is associated with tumor progression. In addition, it has pro-inflammatory functions in endothelial cells and is involved in some inflammatory diseases associated with angiogenesis. Recent research indicated that Tie-1 gene ablation exhibited significant effects on tumor blood- and lymph-angiogenesis and improved anti-Ang therapy, suggesting Tie-1 may be a potential target for tumor anti-angiogenesis treatment.
Angiogenesis Inhibitors ; therapeutic use ; Angiopoietins ; genetics ; metabolism ; Animals ; Embryo, Mammalian ; Embryonic Development ; genetics ; Endothelial Cells ; drug effects ; metabolism ; pathology ; Gene Expression Regulation, Developmental ; Gene Expression Regulation, Neoplastic ; Humans ; Mice ; Neoplasms ; drug therapy ; genetics ; metabolism ; pathology ; Neovascularization, Pathologic ; drug therapy ; genetics ; metabolism ; pathology ; Protein Binding ; Receptor, TIE-1 ; antagonists & inhibitors ; genetics ; metabolism ; Receptor, TIE-2 ; genetics ; metabolism ; Signal Transduction

The purpose of this study is to evaluate the embryotoxicity of alkaloids in Senecionis Scandentis Hebra on in vitro cultured mouse embryos. Mouse whole embryo culture (WEC) was applied in this study. Post-implantation (8.5 d) mouse embryos were isolated from their mothers, and cultured in medium of immediately centrifuged serum (ICS) with different concentrations of seneciphylline (target concentrations were 100, 50, 25 and 12.5 μg x mL(-1)) or senkirkine (target concentrations were 50, 25 and 12.5 μg x mL(-1)) for 48 h. After culturing completed, the development and organic morphodifferentiation of the cultured embryos were evaluated microscopically. Treatment with seneciphylline and senkirkine had adverse effects on the development and organic morphodifferentiation of embryos. The effect also had clear dose-response. Alkaloidals in Senecionis Scandentis Hebra had embryotoxicity on cultured embryos, which indicated that pregnant people exposed to Senecionis Scandentis Hebra may get potential risk on fetus.
Animals ; Embryo Culture Techniques ; Embryo, Mammalian ; drug effects ; Female ; Mice ; Pregnancy ; Pyrrolizidine Alkaloids ; toxicity ; Senecio ; chemistry ; Teratogens ; toxicity

OBJECTIVETo observe the regulatory effects of psoralen, oleanolic acid, and stilbene glucoside, three active components of psoralea fruit, glossy privet fruit and tuber fleeceflower root respectively, on Aβ25-35induced self-renewal and neuron-like differentiation of neural stem cells (NSCs).

METHODSEmbryonic NSCs werein vitro isolated and cultured from Kunming mice of 14-day pregnancy, and randomly divided into the control group, the Aβ25-35 group, the Aβ25-35 +psoralen group, the Aβ25-35 +oleanolic acid group, and the Aβ25-35 + stilbene glucoside group. The intervention concentration of Aβ25-35 was 25 µmol/L, and the intervention concentration of three active components of Chinese medicine was 10(-7)mol/L. The effect of three active components of Chinese medicine on the proliferation of NSCs was observed by counting method. The protein expression of Tubulin was observed by Western blot and immunofluorescence. The ratio of Tubulin+/DAPI was caculated. Results Compared with the control group, the sperical morphology of NSCs was destroyed in the Aβ25-35 group, the counting of NSCs, the expression of Tubulin protein, and the ratio of Tubulin /DAPI all decreased (P <0.01, P <0.05). Compared with the Aβ25-35 group, the counting of NSCs, the expression of Tubulin protein, and the ratio of Tubulin + /DAPI all increased in the three Chinese medicine treated groups (P <0. 01, P <0. 05).

CONCLUSIONS25 µmol/L Aβ25-35 could inhibit self-renewal and neuron-like differentiating of NSCs. But psoralen, oleanolic acid, and stilbene glucoside could promote self-renewal of NSCs and neuron-like differentiation.

Amyloid beta-Peptides ; physiology ; Animals ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Embryo, Mammalian ; Female ; Mice ; Neural Stem Cells ; Neurogenesis ; drug effects ; Neurons ; cytology ; Peptide Fragments ; physiology ; Pregnancy

Insufficient epigenetic reprogramming of donor nuclei is believed to be one of the most important causes of low development efficiency of mammalian somatic cell nuclear transfer (SCNT). Previous studies have shown that both the in vitro and in vivo development of mouse SCNT embryos could be increased significantly by treatment with various histone deacetylase inhibitors (HDACi), including Trichostatin A, Scriptaid, and m-carboxycinnamic acid bishydroxamide (CBHA), in which only the effect of CBHA has not yet been tested in other species. In this paper we examine the effect of CBHA treatment on the development of porcine SCNT embryos. We have discovered the optimum dosage and time for CBHA treatment: incubating SCNT embryos with 2 μmol/L CBHA for 24 h after activation could increase the blastocyst rate from 12.7% to 26.5%. Immunofluorescence results showed that the level of acetylation at histone 3 lysine 9 (AcH3K9), acetylation at histone 3 lysine 18 (AcH3K18), and acetylation at histone 4 lysine 16 (AcH4K16) was raised after CBHA treatment. Meanwhile, CBHA treatment improved the expression of development relating genes such as pou5f1, cdx2, and the imprinted genes like igf2. Despite these promising in vitro results and histone reprogramming, the full term development was not significantly increased after treatment. In conclusion, CBHA improves the in vitro development of pig SCNT embryos, increases the global histone acetylation and corrects the expression of some developmentally important genes at early stages. As in mouse SCNT, we have shown that nuclear epigenetic reprogramming in pig early SCNT embryos can be modified by CBHA treatment.
Acetylation ; Animals ; Blastocyst ; cytology ; Cell Nucleus ; metabolism ; Cinnamates ; pharmacology ; Embryo, Mammalian ; drug effects ; metabolism ; Embryonic Development ; drug effects ; Epigenesis, Genetic ; Female ; Gene Expression ; Histone Deacetylase Inhibitors ; pharmacology ; Histones ; metabolism ; Homeodomain Proteins ; genetics ; metabolism ; In Vitro Techniques ; Insulin-Like Growth Factor II ; genetics ; metabolism ; Nuclear Transfer Techniques ; Octamer Transcription Factor-3 ; genetics ; metabolism ; Swine

This study was conducted to establish an in vitro maturation (IVM) system by selection of efficient porcine serum during porcine in vitro production. To investigate the efficient porcine serum (PS), different types of PS [newborn pig serum, prepubertal gilt serum (PGS), estrus sow serum, and pregnancy sow serum] were used to supplement IVM media with or without gonadotrophin (GTH) and development rates of parthenogenetic activation (PA) and in vitro fertilization (IVF) embryos were then compared. The maturation rates of the PGS group was significantly higher when GTH was not added. Additionally, during development of PA embryos without GTH, the PGS group showed significantly higher cleavage and blastocyst formation rates. Moreover, the cleavage rates of IVF embryos were significantly higher in the PGS group, with no significant differences in the blastocyst formation. However, when GTH was supplemented into the IVM media, there were no significant differences among the four groups in the cleavage rates, development rates of the blastocyst, and cell number of the blastocyst after PA and IVF. In conclusion, PGS is an efficient macromolecule in porcine IVM, and GTH supplementation of the IVM media is beneficial when PS is used as macromolecule, regardless of its origin.
Animals ; Blastocyst/*drug effects ; Embryo, Mammalian/drug effects/*embryology/physiology/ultrastructure ; Fertilization in Vitro/veterinary ; Gonadotropins/administration & dosage/*metabolism ; In Vitro Oocyte Maturation Techniques/*methods/veterinary ; Parthenogenesis/*drug effects ; Sus scrofa/*embryology

OBJECTIVETo investigate the effect of maslinic acid preconditioning against injuries of rat cortical neurons induced by oxygen-glucose deprivation (OGD).

METHODSThe cortical neurons were isolated from Sprague-Dawley rat embryos at 15-17 days of gestation for primary culture. The cortical neurons were incubated with different concentrations (0.1, 1, and 10 micro;mol/L) of maslinic acid prior to OGD. The cell damage and viability were evaluated for lactate dehydrogenase (LDH) leakage and using MTT assay, respectively, and the expression of Bax protein was detected using Western blotting.

RESULTSOGD significantly increased LDH release rate and decreased the viability of the cells. After preconditioning with maslinic acid (1 and 10 micro;mol/L), LDH leakage rate was decreased and cell the viability increased in cells exposed to OGD. Western blotting showed that Bax expression in the cells decreased as maslinic acid concentrations increased.

CONCLUSIONPretreatment with maslinic acid can protect cultured embryonic rat cortical neurons against OGD-induced injury possibly in relation to decreased expression of Bax.

Animals ; Apoptosis ; Cell Hypoxia ; Cells, Cultured ; Cerebral Cortex ; cytology ; drug effects ; Embryo, Mammalian ; Glucose ; metabolism ; L-Lactate Dehydrogenase ; metabolism ; Neurons ; drug effects ; metabolism ; Oxygen ; metabolism ; Rats ; Rats, Sprague-Dawley ; Triterpenes ; pharmacology ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo establish a suitable protocol for activating mouse somatic cell nuclear transfer embryos with strontium chloride (SrCl2).

METHODSWe constructed and identified mouse nuclear transfer (NT) embryos. After nuclear injection, we activated the NT embryos using the following chemical activation methods: exposing the NT embryos to 5 and 10 mmol/L SrCl2 strontium for 1 -8 h, activating the NT embryos with 1-20 mmol/L SrCl2 strontium at 4 and 6 h, treating the NT embryos with 10 mmol/L SrCl2 strontium in different activating media, and exposing the NT embryos to 10 mmol/L SrCl2 strontium combined with 6-dimethylaminopurine (6-DMAP) and cytochalasin B (CB). After activation, the NT embryos were cultured in vitro in the cleavage medium.

RESULTSWhen the NT embryos were treated with SrCl2 at the concentration of 5 mmol/L, the cleavage rate was remarkably higher at 6 h (38.9%) than at 1 h (6.7%), 2 h (22.8%), 3 h (22.8%) and 4 h (25.6%) (P < 0.05), but with no significant differences from those at 5 h (28.9%), 7 h (34.4%) and 8 h (28.9%) (P > 0.05). When the NT embryos were treated with SrCl2 for 6 h, the rates of cleavage and blastulation were 68.9% and 7.2% at 10 mmol/L, markedly higher than at 1 mmol/L (28.3% and 0%), 2.5 mmol/L (35.6% and 0%), 5 mmol/L (37.8% and 1.1%), 7.5 mmol/L (60.6% and 2.2%), 15 mmol/L (51.7% and 1.1%), and 20 mmol/L (41.7% and 1.1%) (P < 0.05). The cleavage rate of the NT embryos cultured in the Ca2+ and Mg2+ KSOM medium was 27.8%, significantly lower than in the Ca(2+)-free KSOM (69.4%), Ca2+/Mg(2+)-free KSOM (66.1%), and Ca2+/Mg(2+)-free + EDTA KSOM (68.3%) (P < 0.05). The total cell blastocyst number was significantly larger in the NT embryos treated with SrCl2 + CB (45.40 +/- 2.23) than in those treated with SrCl2 (30.15 +/- 1.12), 6-DMAP (34.95 +/- 1.38), and 6-DMAP + CB (37.45 +/- 1.43) (P < 0.05).

CONCLUSIONSix-hour treatment with 10 mmol/L SrCl2 in Ca2+ alone or in combination with CB can well activate NT embryos in mice.

Animals ; Blastocyst ; cytology ; drug effects ; Embryo Culture Techniques ; Embryo, Mammalian ; cytology ; drug effects ; Embryonic Development ; drug effects ; Female ; Mice ; Mice, Inbred C57BL ; Nuclear Transfer Techniques ; Oocytes ; cytology ; drug effects ; Strontium ; pharmacology

OBJECTIVETo investigate the fetotoxicity of retrorsine.

METHODMouse whole embryo culture (WEC) was applied. Post-implantation (8.5 d) mouse embryos were isolated from their mothers and put into the medium of immediately centrifuged serum (ICS) prepared from rats. Different concentrations of retrorsine (12.5, 25, 50, 100 mg x L(-1)) were added into the WEC culture. Development (yolk sac diameter, crown-rump length, head length, somite number) and organic morphodifferentiation (yolk sac circulation, allantois, embryonic flexion, heart, brain, optic-otic-olfactory organ, branchial arch, maxillary, mandible, bud) of embryos were observed at 48 h after treatment.

RESULTObvious fetotoxicity could be observed in various retrorsine treatment groups in a dose-dependent manner. Development of embryos was delayed significantly at dose 12.5-100 mg x L(-1). Malformations were shown in all organic morphodifferentiation indexes, especially in otic-olfactory organ, branchial arch, maxillary, mandible, bud.

CONCLUSIONRetrorsine had obvious fetotoxicity in vitro WEC culture, indicating that exposure of pregnant mice to retrorsine may have potential risk on fetals.

Animals ; Dose-Response Relationship, Drug ; Embryo, Mammalian ; drug effects ; Female ; Male ; Mice ; Pregnancy ; Pyrrolizidine Alkaloids ; toxicity ; Rats ; Toxicity Tests ; methods