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Zygote: the Biology of Gametes and Early Embryos

Soleiman Kurd, Sara Hosseini, Fardin Fathi, Vahid Jajarmi, Mohammad Salehi
SummaryOne of the methods to generate transgenic animals is called sperm-mediated gene transfer (SMGT). Mature sperm cells can take up exogenous DNA molecules intrinsically and transfer them into the oocyte during fertilization. This study assessed the effect of dimethyl sulfoxide (DMSO) and electrolyte-free medium (EFM) on DNA uptake (EGFP-N1plasmid) in mouse sperm. Sperms cells cultured in human tubular fluid (HTF) without any treatment were considered as the control group. Sperms cells that were incubated in EFM and HTF with DNA/DMSO at 4°C were classified into EFM and HTF groups...
October 31, 2018: Zygote: the Biology of Gametes and Early Embryos
Feo Lima, Ftg Bezerra, G B Souza, Mht Matos, R van den Hurk, Jrv Silva
SummaryThis study aimed to investigate the effects of IL1β and TNFα on growth and maturation of oocytes from small follicles (1-3 mm) during in vitro culture. To this end, cumulus-oocyte complexes (COCs) with diameters of ~110 µm were cultured in TCM-199 medium alone or supplemented with IL1β (10 ng/ml), TNFα (10 ng/ml) or both for 48 h. The oocytes were measured at the beginning and at the end of the culture period. COCs were cultured for 20 h in pre-maturation medium and then half of the COCs of each group was destined for in vitro maturation and the remaining COCs were used to evaluate meiotic progression, mitochondrial distribution and the expression of mRNAs for GDF-9, c-Mos, Cyclin-B1 and H1foo...
October 31, 2018: Zygote: the Biology of Gametes and Early Embryos
George Shigueki Yasui, Taiju Saito, Yan Zhao, Takafumi Fujimoto, Etsuro Yamaha, Katsutoshi Arai
SummaryPolyspermy was initiated by microinjecting a multiple number of sperm into the activated and dechorionated eggs of dojo loach Misgurnus anguillicaudatus (Teleostei: Cobitidae). A 10-nl sperm suspension from an albino (recessive trait) male (105, 106, 107 or 108 sperm ml -1) was microinjected into eggs from a wild-type female. Although the rates of embryos developing into the blastula stage in the injection group at the highest sperm concentration were similar to that of the control group, the hatching rates of the injection group were much lower...
October 29, 2018: Zygote: the Biology of Gametes and Early Embryos
Martin Morovic, Olga Østrup, Frantisek Strejcek, Michal Benc, Matej Murin, Katarina Jedlickova, Alexandra Bartkova, Andrea Lucas-Hahn, Lazo Pendovski, Jozef Laurincik
SummaryThe present study examines the role of RNA polymerase I (RPI)-mediated transcription, maternally inherited rRNA and nucleolar proteins in the resumption of fibrillogranular nucleoli during embryonic genome activation (EGA) in porcine embryos. Late 4-cell embryos were incubated in the absence (control) or presence of actinomycin D (AD) (0.2 μg/ml for inhibition of RPI; 2.0 μg/ml for inhibition of total transcription) and late 2-cell embryos were cultured to the late 4-cell stage with 0.2 μg/ml AD to block EGA...
October 12, 2018: Zygote: the Biology of Gametes and Early Embryos
A Hassan, V T Okomoda, F A B Sanusi
SummaryThis study investigated the breeding parameters and embryogenic development of diploid and heat shock-induced triploid eggs of Anabas testudineus (Bloch, 1792). To this effect, broodstocks of A. testudineus were induced to spawn using the Ovaprim® hormone. After fertilization, the eggs were divided into two groups and one portion heat shocked at 41°C (for 3 min), at approximately 4 min after fertilization. Results of fertilization, hatchability, as well as the sequence and timing of embryogenic development were collated from three breeding trials...
October 9, 2018: Zygote: the Biology of Gametes and Early Embryos
Marcela Pinheiro Paz, Francisca Geovania Canafístula de Sousa, Benner Geraldo Alves, Carlos Henrique Lobo, Antonia Débora Sales, Kaio Cesar Simiano Tavares, Naíza Arcângela Ribeiro de Sá, Denise Damasceno Guerreiro, Carolina Maside, Rebeca Magalhães Pedrosa Rocha, Luciana Relly Bertolini, Vilceu Bordignon, José Ricardo de Figueiredo, Ana Paula Ribeiro Rodrigues
SummaryThe objectives were to develop an effective protocol for transfection of ovine secondary follicles and to assess the effect of attenuating aquaporin 3 (AQP3) using a small interfering RNA (siRNA-AQP3) on antrum formation and follicular growth in vitro. Various combinations of Lipofectamine® volumes (0.5, 0.75 or 1.0 µl), fluorescent oligonucleotide (BLOCK-iT ™) concentrations (3.18, 27.12 or 36.16 nM) and exposure times (12, 14, 16, 18 or 20 h) were tested. The BLOCK-iT™ was replaced by siRNA-AQP3 in the transfection complex...
October 5, 2018: Zygote: the Biology of Gametes and Early Embryos
Atsushi Saitoh, Yuki Takada, Mayu Horie, Tomoya Kotani
SummaryTranslational regulation of mRNAs is crucial for promoting various cellular and developmental processes. Pumilio1 (Pum1) has been shown to play key roles in translational regulation of target mRNAs in many systems of diverse organisms. In zebrafish immature oocytes, Pum1 was shown to bind to cyclin B1 mRNA and promote the formation of cyclin B1 RNA granules. This Pum1-mediated RNA granule formation seemed critical to determine the timing of translational activation of cyclin B1 mRNA during oocyte maturation, leading to activation of maturation/M-phase-promoting factor (MPF) at the appropriate timing...
October 5, 2018: Zygote: the Biology of Gametes and Early Embryos
Manish Kumar, Mona Faraji, Parul Sarwalia, Sandeep Kumar, Moloya Gohain, Sachinandan De, Rakesh Kumar, T K Datta
SummaryMaturing oocytes have diverse developmental potential and good quality oocytes exhibit a better ability to attain physiological milestones in a time-dependent manner. This situation necessitates the confirmation of oocyte developmental status more precisely under an in vitro embryo production (IVEP) regime. The aim of this study was to explain timely events in germinal vesicle breakdown (GVBD), an important milestone of oocyte nuclear maturation, to delineate the developmental capacity of Bubalus bubalis oocytes...
October 5, 2018: Zygote: the Biology of Gametes and Early Embryos
Thanapol Nongbua, Essraa M Al-Essawe, Anders Edman, Anders Johannisson, Jane M Morrell
SummaryThe aim of this study was to investigate the effect of adding homologous or heterologous bovine seminal plasma (SP) to SP-free sperm samples before freezing on sperm quality after thawing. Ejaculates from bulls of known fertility were used as a source of SP. The SP was removed from further aliquots of the same ejaculates by colloid centrifugation to create SP-free sperm samples; the resuspended sperm pellets were treated with homologous or heterologous SP from high or low fertility bulls at 0%, 1% or 5% before freezing...
October 5, 2018: Zygote: the Biology of Gametes and Early Embryos
Elnaz Zand, Rouhollah Fathi, Mitra Heydari Nasrabadi, Mohammad Jafari Atrabi, Norah Spears, Vahid Akbarinejad
SummaryThe high miscarriage rates that result following transfer of embryos derived from in vitro maturation (IVM) of oocytes necessitate improvements in the processes involved. This study aimed to improve the quality of in vitro matured oocytes using granulosa cell conditioned medium (GCCM) as the culture medium. In this work, germinal vesicle (GV)-stage oocytes from NMRI mice were collected and cultured using three types of culture medium: Base medium (BM) (control), 50% granulosa cell conditioned medium (GCCM50) and 100% GCCM (GCCM100)...
October 3, 2018: Zygote: the Biology of Gametes and Early Embryos
Ahmed M Elomda, Mohamed F Saad, Ayman M Saeed, Ashraf Elsayed, Ahmed O Abass, Hosam M Safaa, Gamal M K Mehaisen
SummaryOxidative stress is a major cause of defective embryo development during in vitro culture. Retinoids are recognized as non-enzymatic antioxidants and may have an important role in the regulation of cell differentiation and vertebrate development. However, there are not enough reports discussing the antioxidant and developmental capacity of retinoids, including retinol (RT), on the in vitro development of embryos recovered from livestock animals, particularly in rabbit species. Therefore, morula embryos obtained from nulliparous Red Baladi rabbit does were cultured for 48 h in TCM199 medium in the absence of RT (control group) or in the presence of RT at concentrations of 10, 100 and 1000 nM...
August 2018: Zygote: the Biology of Gametes and Early Embryos
Tuğba Kotil, M Ertan Kervancıoğlu, Gülçin Ekter Kanten, Gülden Tunalı, Seyhun Solakoğlu
SummaryDigyny, the presence of a third pronucleus due to the failure of second polar body extrusion, is problematic after intracytoplasmic sperm injection (ICSI) practices. Mitochondria have critical roles such as production of adenosine triphosphate (ATP) and regulation of Ca2+ homeostasis during oocyte maturation, fertilization and the following development, while the regulation of meiotic spindle formation, chromosome segregation, pronuclear apposition and cytokinesis is closely associated with the cytoskeleton...
August 2018: Zygote: the Biology of Gametes and Early Embryos
D V C Almeida, J S Lima, D L Leão, K G Oliveira, R R Santos, S F S Domingues, M S Miranda
SummaryThe aim of this study was to evaluate the effect of incubating semen for different periods (90, 270 or 450 min) with or without Trolox® (100 or 150 µM) on the quality of sperm from Saimiri collinsi. Sperm motility, vigour, and plasma membrane integrity (PMI) were evaluated in both fresh semen and semen incubated for different time periods, i.e. 90, 270 or 450 min of incubation. Supplementation of semen extender with Trolox® 100 µM improved sperm motility, vigour and PMI for up to 270 min of incubation...
August 2018: Zygote: the Biology of Gametes and Early Embryos
Frederico da Silva Bitecourt, Carina Oliveira Dumont Horta, Karen Santos Lima, Bruno Bastos Godoi, Fernanda Luiza Menezes Bello, Cíntia Maria Rodrigues, Luana Pereira Leite Schetino, Kinulpe Honorato-Sampaio
SummaryPrevious studies have established a model of atresia in preovulatory follicles after stimulation of immature rats with equine chorionic gonadotropin (eCG). This gonadotropin recruits a follicular pool and the deprivation of preovulatory luteinizing hormone (LH) surge induces the atresia in preovulatory follicles. The present study investigated the occurrence of ovulation and provided some morphological features of granulosa cell (GC) apoptosis of atretic follicles at 0, 48, 72 and 120 h after eCG stimulation...
August 2018: Zygote: the Biology of Gametes and Early Embryos
João Diego de Agostini Losano, Daniel de Souza Ramos Angrimani, Roberta Ferreira Leite, Bárbara do Carmo Simões da Silva, Valquíria Hyppolito Barnabe, Marcilio Nichi
SummaryDespite sperm mitochondrial relevance to the fertilization capacity, the processes involved in the production of ATP and functional dynamics of sperm mitochondria are not fully understood. One of these processes is the paradox involved between function and formation of reactive oxygen species performed by the organelle. Therefore, this review aimed to provide data on the role of sperm mitochondria in oxidative homeostasis and functionality as well the tools to assess sperm mitochondrial function.
August 2018: Zygote: the Biology of Gametes and Early Embryos
Luciana Nakaghi Ganeco-Kirschnik, Irene Bastos Franceschini-Vicentini, Maria do Carmo Faria Paes, Laura Satiko Okada Nakaghi
Brycon orbignyanus is an important large teleost that is currently on the list of endangered species, therefore studies on its reproductive biology and embryology are fundamental to help species conservation and recovery. The objective of this research was to characterize the events that occur during extrusion, fertilization and embryonic development of the species. The samples were collected at predetermined times, fixed and processed for light microscopy and scanning electron microscopy. The greenish oocytes were spherical, had translucent chorion and a mean diameter of 1...
August 2018: Zygote: the Biology of Gametes and Early Embryos
Rosilane Gomes de Souza de Oliveira, Arley Faria José de Oliveira, Marle Angélica Villacorta Correa
SummaryThe aim of this study was to evaluate if hormonal treatment causes changes in the morphology of matrinxã Brycon amazonicus spermatozoa. Twelve males were randomized into three treatment groups: 1, Ovaprim™ (0.5 ml/kg); 2, carp pituitary extract (CPE; 2.0 mg/kg); and 3, NaCl solution 0.9% - control group (0.5 ml/kg); with four replicates. Morphological sperm analysis was performed using an optical and scanning electron microscope. The percentage of normal spermatozoa (49.6±4.6% to 60.8±2.8%), with primary (26...
August 2018: Zygote: the Biology of Gametes and Early Embryos
Seda Karabulut, Asuman Demiroğlu-Zergeroğlu, Elif Yılmaz, Pelin Kutlu, İlknur Keskin
SummaryThe negative effects of cryopreservation on sperm parameters are well documented but little information is known about molecular basis of the process. The aim of the present study was to investigate the possible effects of sperm cryopreservation on main apoptotic signs including DNA fragmentation and caspase-3 activation and to determine if these effects vary according to sperm parameters. Sperm samples of 72 patients were cryopreserved. The patients were sub-grouped as normozoospermic or non-normozoospermic patients according to their semen parameters...
August 2018: Zygote: the Biology of Gametes and Early Embryos
José A B Bezerra, Andréia M Silva, Patrícia C Sousa, Lívia B Campos, Érica C G Praxedes, Luana G P Bezerra, Thibério S Castelo, Ana L P Souza, Alexandre R Silva
SummaryThe aim of this study was to establish a functional freezing-thawing protocol for epididymal sperm of collared peccaries (Pecari tajacu L., 1758) by comparing different extenders. The epididymal sperm from 12 sexually mature males was recovered by retrograde flushing using Tris-based or coconut water-based (ACP®-116c) extenders. After initial evaluation, samples were diluted and frozen with the same extenders to which 20% egg yolk and 6% glycerol were added. After 2 weeks, thawing was performed at 37°C/60 s and sperm motility, vigour, morphology, functional membrane integrity, sperm viability, sperm plasma membrane integrity, and a computer-assisted semen analysis (CASA) were assessed...
August 2018: Zygote: the Biology of Gametes and Early Embryos
Zigomar da Silva, Andressa Pereira de Souza, José Rodrigo Claudio Pandolfi, Francisco Noé da Fonseca, Carlos André da Veiga Lima-Rosa, Mariana Groke Marques
SummaryThe aim of this study was to optimize protocols for electroporation (EP) and polyfection (PLF) using polyethyleneimine (PEI) for pig sperm transfection and to determine which method was the most efficient. For EP standardization, different voltages, amounts and times of electric pulses were tested using propidium iodide (PI) as reporter. For PLF standardization, different concentrations of fluorescein isothiocyanate (FITC)-labelled PEI (PEI/FITC) were incubated with sperm for different periods of time...
August 2018: Zygote: the Biology of Gametes and Early Embryos
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