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the nucleus of the ovum or sperm after fertilization but before they fuse to form the nucleus of the zygote

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出典(authority):フリー百科事典『ウィキペディア（Wikipedia）』「2016/01/23 10:33:55」(JST)

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1. 卵細胞質内精子注入法 intracytoplasmic sperm injection

English Journal

A non-surgical approach for male germ cell mediated gene transmission through transgenesis.

Usmani A, Ganguli N, Sarkar H, Dhup S, Batta SR, Vimal M, Ganguli N, Basu S, Nagarajan P, Majumdar SS.Author information Embryo Biotechnology Laboratory, National Institute of Immunology, New Delhi, India.AbstractMicroinjection of foreign DNA in male pronucleus by in-vitro embryo manipulation is difficult but remains the method of choice for generating transgenic animals. Other procedures, including retroviral and embryonic stem cell mediated transgenesis are equally complicated and have limitations. Although our previously reported technique of testicular transgenesis circumvented several limitations, it involved many steps, including surgery and hemicastration, which carried risk of infection and impotency. We improved this technique further, into a two step non-surgical electroporation procedure, for making transgenic mice. In this approach, transgene was delivered inside both testes by injection and modified parameters of electroporation were used for in-vivo gene integration in germ cells. Using variety of constructs, germ cell integration of the gene and its transmission in progeny was confirmed by PCR, slot blot and immunohistochemical analysis. This improved technique is efficient, requires substantially less time and can be easily adopted by various biomedical researchers.
Scientific reports.Sci Rep.2013 Dec 5;3:3430. doi: 10.1038/srep03430.
Microinjection of foreign DNA in male pronucleus by in-vitro embryo manipulation is difficult but remains the method of choice for generating transgenic animals. Other procedures, including retroviral and embryonic stem cell mediated transgenesis are equally complicated and have limitations. Althoug
PMID 24305437

Comparison of cytoskeletal integrity, fertilization and developmental competence of oocytes vitrified before or after in vitro maturation in a porcine model.

Egerszegi I, Somfai T, Nakai M, Tanihara F, Noguchi J, Kaneko H, Nagai T, Rátky J, Kikuchi K.Author information Research Institute for Animal Breeding and Nutrition, Herceghalom H-2053, Hungary. Electronic address: istvan.egerszegi@atk.hu.AbstractAim of the study was to investigate the effect of vitrification on viability, cytoskeletal integrity and in vitro developmental competence after in vitro fertilization (IVF) of oocytes vitrified before or after in vitro maturation (IVM) using a pig model. Oocytes from abattoir-derived porcine ovaries were vitrified at either the germinal vesicle (GV) or metaphase II (MII) stage by modified solid surface vitrification (SSV). Oocyte viability was evaluated by stereomicroscopic observation whereas their nuclear stage and morphology of microtubules and F-actin were observed by confocal microscopy after immunostaining. Fertilization was assessed by orcein staining. The survival rate after vitrification was higher for MII-stage than for GV-stage oocytes. However, the ability of surviving oocytes to reach the MII stage after vitrification at the GV stage (GV-vitrified oocytes) was similar to that of control oocytes. Furthermore, after IVM, GV-vitrified oocytes had better spindle and F-actin integrity than oocytes vitrified at the MII stage (MII-vitrified oocytes). In accordance with this result, GV-vitrified oocytes had better ability to extrude the second polar body and support male pronucleus formation after in vitro fertilization (IVF), in comparison to MII-vitrified oocytes. Fertilization rates did not differ among groups. Finally, the ability of GV-vitrified oocytes to develop into embryos was superior to that of MII-vitrified oocytes. However, both vitrified groups showed reduced blastocyst development compared with the control group. In conclusion vitrification of porcine oocytes at the GV stage is advantageous in conferring better cytoskeletal organization and competence to develop to the blastocyst stage in comparison with vitrification at the MII stage.
Cryobiology.Cryobiology.2013 Dec;67(3):287-92. doi: 10.1016/j.cryobiol.2013.08.009. Epub 2013 Aug 28.
Aim of the study was to investigate the effect of vitrification on viability, cytoskeletal integrity and in vitro developmental competence after in vitro fertilization (IVF) of oocytes vitrified before or after in vitro maturation (IVM) using a pig model. Oocytes from abattoir-derived porcine ovarie
PMID 23993921

Active DNA demethylation in post-mitotic neurons: A reason for optimism.

Gavin DP1, Chase KA2, Sharma RP3.Author information 1The Psychiatric Institute, Department of Psychiatry, University of Illinois at Chicago, 1601 W. Taylor St., Chicago, IL 60612, USA; Jesse Brown Veterans Affairs Medical Center, 820 South Damen Avenue (M/C 151), Chicago, IL 60612, USA. Electronic address: dgavin@psych.uic.edu.2The Psychiatric Institute, Department of Psychiatry, University of Illinois at Chicago, 1601 W. Taylor St., Chicago, IL 60612, USA.3The Psychiatric Institute, Department of Psychiatry, University of Illinois at Chicago, 1601 W. Taylor St., Chicago, IL 60612, USA; Jesse Brown Veterans Affairs Medical Center, 820 South Damen Avenue (M/C 151), Chicago, IL 60612, USA.AbstractOver the last several years proteins involved in base excision repair (BER) have been implicated in active DNA demethylation. We review the literature supporting BER as a means of active DNA demethylation, and explain how the various components function and cooperate to remove the potentially most enduring means of epigenetic gene regulation. Recent evidence indicates that the same pathways implicated during periods of widespread DNA demethylation, such as the erasure of methyl marks in the paternal pronucleus soon after fertilization, are operational in post-mitotic neurons. Neuronal functional identities, defined here as the result of a combination of neuronal subtype, location, and synaptic connections are largely maintained through DNA methylation. Chronic mental illnesses, such as schizophrenia, may be the result of both altered neurotransmitter levels and neurons that have assumed dysfunctional neuronal identities. A limitation of most current psychopharmacological agents is their focus on the former, while not addressing the more profound latter pathophysiological process. Previously, it was believed that active DNA demethylation in post-mitotic neurons was rare if not impossible. If this were the case, then reversing the factors that maintain neuronal identity, would be highly unlikely. The emergence of an active DNA demethylation pathway in the brain is a reason for great optimism in psychiatry as it provides a means by which previously pathological neurons may be reprogrammed to serve a more favorable role. Agents targeting epigenetic processes have shown much promise in this regard, and may lead to substantial gains over traditional pharmacological approaches.
Neuropharmacology.Neuropharmacology.2013 Dec;75:233-45. doi: 10.1016/j.neuropharm.2013.07.036. Epub 2013 Aug 16.
Over the last several years proteins involved in base excision repair (BER) have been implicated in active DNA demethylation. We review the literature supporting BER as a means of active DNA demethylation, and explain how the various components function and cooperate to remove the potentially most e
PMID 23958448

Japanese Journal

Treating Cloned Embryos, but Not Donor Cells, with 5-aza-2-deoxycytidine Enhances the Developmental Competence of Porcine Cloned Embryos

HUAN Yan Jun,ZHU Jiang,XIE Bing Teng,WANG Jian Yu,LIU Shi Chao,ZHOU Yang,KONG Qing Ran,HE Hong Bin,LIU Zhong Hua
Journal of Reproduction and Development, 2013
… Therefore, in this study, we treated porcine fetal fibroblasts (PFF) that then were used as donor nuclei for nuclear transfer or fibroblast-derived reconstructed embryos with 5-aza-dC, and the concentration- and time-dependent effects of 5-aza-dC on porcine cloned embryos were investigated by assessing pseudo-pronucleus formation, developmental potential and pluripotent gene expression of these reconstructed embryos. …
NAID 130003361044

In Vitro Fertilization and Development of Porcine Oocytes Matured in Follicular Fluid

AGUNG Budiyanto,OTOI Takeshige,FUCHIMOTO Dai-ichiro,SENBON Shoichiro,ONISHI Akira,NAGAI Takashi
Journal of Reproduction and Development 59(2), 103-106, 2013
… The total mean rates of sperm penetration, normal fertilization, male pronucleus (MPN) formation, cleavage, and development to the blastocyst stage of oocytes after insemination were significantly higher (P<0.01) in the rotating culture system than in the static culture system. …
NAID 130003361012

Sperm Pretreatment with Dithiothreitol Increases Male Pronucleus Formation Rates After Intracytoplasmic Sperm Injection (ICSI) in Swamp Buffalo Oocytes

CHANKITISAKUL Vibuntita,AM-IN Nutthee,THARASANIT Theerawat,SOMFAI Tamas,NAGAI Takashi,TECHAKUMPHU Mongkol
Journal of Reproduction and Development 59(1), 66-71, 2013
… Failure of male pronucleus formation has hampered the success of intracytoplasmic sperm injection (ICSI) in swamp buffalo. … The aim of the present study was to improve male pronucleus formation by pretreating sperm with various chemicals before ICSI. … At 18 h post-ICSI, pronucleus (PN) formation was found only in activated oocytes. …
NAID 130001854906