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Nathalie Jiatsa Donfack, Kele Amaral Alves, Benner Geraldo Alves, Rebeca Magalhães Pedrosa Rocha, Jamily Bezzera Bruno, Marcelo Bertolini, Regiane Rodrigues Dos Santos, Sheyla Farhaydes Souza Domingues, José Ricardo De Figueiredo, Johan Smitz, Ana Paula Ribeiro Rodrigues
This study aimed to evaluate the follicular morphology and development (follicular activation, cell proliferation, and hormone production), as well as the distribution pattern of Connexins 37 and 43 and SDF-1α after vitrification and in vitro culture of goat ovarian tissue. The study involved four experimental groups: fresh control, vitrified control, fresh culture and vitrified culture. The ovarian fragments were vitrified by a solid surface technique using the Ovarian Tissue Cryosystem and subsequently in vitro cultured for 7 days...
May 12, 2018: Theriogenology
Cristina Matthewman, Christina K Johnson, David M Miller Iii, Laura Bianchi
UNC-8 and MEC-4 are two members of the DEG/ENaC family of voltage-independent Na+ channels that share a high degree of sequence homology and functional similarity. For example, both can be hyperactivated by genetic mutations (UNC-8(d) and MEC-4(d)) that induce neuronal death by necrosis. Both depend in vivo on chaperone protein MEC-6 for function, as demonstrated by the finding that neuronal death induced by hyperactive UNC-8 and MEC-4 channels is prevented by null mutations in mec-6. UNC-8 and MEC-4 differ functionally in three major ways: 1) MEC-4 is calcium permeable whereas UNC-8 is not; 2) UNC-8, but not MEC-4, is blocked by extracellular calcium and magnesium in the micromolar range; 3) MEC-6 increases the number of MEC-4 channels at the cell surface in oocytes but does not have this effect on UNC-8...
April 25, 2018: American Journal of Physiology. Cell Physiology
Teppei Nakamura, Masataka Chihara, Osamu Ichii, Saori Otsuka-Kanazawa, Ken-Ichi Nagasaki, Yaser Hosny Ali Elewa, Osamu Tatsumi, Yasuhiro Kon
MRL/MpJ mice have abundant ovarian mast cells (MCs) as compared with other strains at postnatal day 0 (P0); however, they sharply decrease after birth. These ovarian MCs, particularly beneath the ovarian surface epithelium (SE), which express mucosal MC (MMC) marker, might participate in early follicular development. This study investigated the changes in spatiotemporal distribution of MCs in the perinatal MRL/MpJ mouse ovaries. At P0 to P7, the MCs were densely localized to the ovary, especially their caudomedial region around the ovary-fimbria connection...
2018: PloS One
Beatriz Fernandez-Fuertes, Alfonso Blanco-Fernandez, Colm J Reid, Kieran Meade, S Fair, Pat Lonergan
This study tested the hypothesis that sperm sialic acid (Sia) is required to reach the site of fertilization, and that successful fertilization requires recognition of Sia from both the sperm and oocyte to occur. In addition, it has recently been reported that Siglecs (Sia-binding-immunoglobulin-like lectins) are present on the sperm surface. Thus, the possibility that the recognition of oocyte Sia was sperm-Siglec-mediated was also addressed. Sperm exposed to neuraminidase (NMase) exhibited lower overall and progressive motility, which translated to a decreased ability to swim through cervical mucus from cows in oestrus...
April 4, 2018: Reproduction: the Official Journal of the Society for the Study of Fertility
Sira Defaus, Manuel Avilés, David Andreu, Ricardo Gutiérrez-Gallego
Seminal plasma proteins are relevant for sperm functionality and some appear responsible for establishing sperm interactions with the various environments along the female genital tract towards the oocyte. In recent years, research has focused on characterizing the role of these proteins in the context of reproductive biology, fertility diagnostics and treatment of related problems. Herein, we focus on the main protein of bovine seminal plasma, PDC-109 (BSP-A1/-A2), which by virtue of its lectin properties is involved in fertilization...
April 4, 2018: International Journal of Molecular Sciences
K Alkhodair, H Almhanna, J McGetrick, S Gedair, M E Gallagher, B Fernandez-Fuertes, T Tharmalingam, P B Larsen, E Fitzpatrick, P Lonergan, A C O Evans, S D Carrington, C J Reid
Sialic acid (Sia) is a major constituent of both the sperm glycocalyx and female reproductive mucosal surface and is involved in regulating sperm migration, uterotubal reservoir formation and oocyte binding. Siglecs (sialic acid-binding immunoglobulin - like lectins) commonly found on immune cells, bind to Sia in a linkage- and sugar-specific manner and often mediate cell-to-cell interactions and signalling. Proteomic and transcriptomic analysis of human and bovine sperm have listed Siglecs, but to date, their presence and/or localisation on sperm has not been studied...
April 2018: Reproduction: the Official Journal of the Society for the Study of Fertility
Lukas Wichmann, Kirsty Sophia Vowinkel, Alexander Perniss, Ivan Manzini, Mike Althaus
The epithelial sodium channel (ENaC) is a critical regulator of vertebrate electrolyte homeostasis. ENaC is the only constitutively open ion channel in the degenerin/ENaC protein family, and its expression, membrane abundance, and open probability therefore are tightly controlled. The canonical ENaC is composed of three subunits (α, β and γ), but a fourth δ-subunit may replace α and form atypical δβγ-ENaCs. Using Xenopus laevis as a model, here we found that mRNAs of the α- and δ-subunits are differentially expressed in different tissues and that δ-ENaC predominantly is present in the urogenital tract...
March 25, 2018: Journal of Biological Chemistry
Stephany El-Hayek, Qin Yang, Laleh Abbassi, Greg FitzHarris, Hugh J Clarke
Germ cells develop in a microenvironment created by the somatic cells of the gonad [1-3]. Although in males, the germ and somatic support cells lie in direct contact, in females, a thick extracellular coat surrounds the oocyte, physically separating it from the somatic follicle cells [4]. To bypass this barrier to communication, narrow cytoplasmic extensions of the follicle cells traverse the extracellular coat to reach the oocyte plasma membrane [5-9]. These delicate structures provide the sole platform for the contact-mediated communication between the oocyte and its follicular environment that is indispensable for production of a fertilizable egg [8, 10-15]...
March 8, 2018: Current Biology: CB
JoAnne S Richards
Follicular development and ovulation are complex development processes that are regulated by multiple, interacting pathways and cell types. The oocyte, cumulus cells, granulosa cells, and theca cells communicate to impact follicular development and ovulation. Many hormones and cytokines control intracellular regulatory networks and transcription factors, some of which are cell type specific. Molecular biology approaches and mutant mouse models have contributed immensely to our knowledge of what genes and signaling cascades impact each stage of follicular development and ovulation, and how the alteration of gene expression profiles and the activation of specific signaling pathways can impact ovarian cancer development in ovarian surface epithelial cells as well as granulosa cells...
2018: Vitamins and Hormones
Mohamed Fathi, Adel R Moawad, Magdy R Badr
Cryopreservation of oocytes would serve as an alternative to overcome the limited availability of dromedary camel oocytes and facilitate improvements in IVP techniques in this species. Our goal was to develop a protocol for the vitrification of camel oocytes at the germinal vesicle (GV) stage using different cryoprotectant combinations: 20% EG and 20% DMSO (VS1), 25% EG plus 25% DMSO (VS2) or 25% EG and 25% glycerol (VS3) and various cryo-carriers; straws or open pulled-straw (OPS) or solid surface vitrification (SSV); and Cryotop...
2018: PloS One
Jessica Kabutomori, Olivia Beloto-Silva, R Ryan Geyer, Raif Musa-Aziz
Xenopus laevis oocytes are a valuable tool for investigating the function of membrane proteins. However, regulations around the world, specifically in Brazil, render the import of Xenopus laevis frogs impractical, and, in some cases, impossible. Here, as an alternative, we evaluate the usefulness of the North American aquatic bullfrog Lithobates catesebeianus , which is commercially available in Brazil, for the heterologous expression of aquaporin (AQP) proteins. We have developed a method that combines a brief collagenase treatment and mechanical defolliculation for isolating individual oocytes from Lithobates ovaries...
March 29, 2018: Biology Open
Md Fazlur Rahman, Radhika Raj, Rajgopal Govindarajan
Combination antiretroviral drug treatments depend on 3'-deoxy-nucleoside analogs such as 3'-azido-3'-deoxythymidine (AZT) and 2'3'-dideoxyinosine (DDI). Despite being effective in inhibiting human immunodeficiency virus replication, these drugs produce a range of toxicities, including myopathy, pancreatitis, neuropathy, and lactic acidosis, that are generally considered as sequelae to mitochondrial damage. Although cell surface-localized nucleoside transporters, such as human equilibrative nucleoside transporter 2 (hENT2) and human concentrative nucleoside transporter 1 (hCNT1), are known to increase the carrier-mediated uptake of 3'-deoxy-nucleoside analogs into cells, another ubiquitously expressed intracellular nucleoside transporter (namely, hENT3) has been implicated in the mitochondrial transport of 3'-deoxy-nucleoside analogs...
May 2018: Drug Metabolism and Disposition: the Biological Fate of Chemicals
Kiley A Knapp, Eusebio S Pires, Sara J Adair, Arabinda Mandal, Anne M Mills, Walter C Olson, Craig L Slingluff, J Thomas Parsons, Todd W Bauer, Timothy N Bullock, John C Herr
Successful therapeutic options remain elusive for pancreatic cancer. The exquisite sensitivity and specificity of humoral and cellular immunity may provide therapeutic approaches if antigens specific for pancreatic cancer cells can be identified. Here we characterize SAS1B (ovastacin, ASTL , astacin-like), a cancer-oocyte antigen, as an attractive immunotoxin target expressed at the surface of human pancreatic cancer cells, with limited expression among normal tissues. Immunohistochemistry shows that most pancreatic cancers are SAS1Bpos (68%), while normal pancreatic ductal epithelium is SAS1Bneg ...
February 6, 2018: Oncotarget
S Haerteis, A Schork, T Dörffel, B N Bohnert, R Nacken, M Wörn, M Xiao, D Essigke, A Janessa, A H Schmaier, E P Feener, H-U Häring, M Bertog, C Korbmacher, F Artunc
AIM: Recent work has demonstrated that activation of the epithelial sodium channel (ENaC) by aberrantly filtered serine proteases causes sodium retention in nephrotic syndrome. The aim of this study was to elucidate a potential role of plasma kallikrein (PKLK) as a candidate serine protease in this context. METHODS: We analysed PKLK in the urine of patients with chronic kidney disease (CKD, n = 171) and investigated its ability to activate human ENaC expressed in Xenopus laevis oocytes...
February 28, 2018: Acta Physiologica
Youmei Wu, María José Luna, Lauren S Bonilla, Nicholas J P Ryba, James M Pickel
Design and engineering of complex knockin mice has revolutionized the in vivo manipulation of genetically defined cells. Recently development of the bacterial clustered regularly interspersed short palindromic repeats (CRISPR) associated protein 9 (Cas9) system for single site cleavage of mammalian genomes has opened the way for rapid generation of knockin mice by targeting homology directed repair to selected cleavage sites. We used this approach to generate new lines of mice that will be useful for a variety of aspects of neuroscience research...
2018: PloS One
J D J Gilbert, A Wells, S J Simpson
Costs and benefits of group living are a fundamental topic in behavioural ecology. Resource availability affects individuals' breeding prospects alone and in groups, as well as how reproduction is distributed within groups ("reproductive skew"). Here, in facultatively social thrips, we provide correlational evidence that breeding resources are associated with (1) whether solitary or social living is favoured, and (2) the degree of ovarian skew. Dunatothrips aneurae (Thysanoptera, Phlaeothripidae) cooperatively build silk "domiciles" on Australian Acacias, feeding exclusively from internal phyllode surfaces...
February 26, 2018: Scientific Reports
Damià Castelló, Ana Cobo, Enric Mestres, Maria Garcia, Ivette Vanrell, José Alejandro Remohí, Gloria Calderón, Nuno Costa-Borges
Vitrification is currently a well-established technique for the cryopreservation of oocytes and embryos. It can be achieved either by direct (open systems) or indirect (closed systems) contact with liquid nitrogen. While there is not a direct evidence of disease transmission by transferred cryopreserved embryos, it was experimentally demonstrated that cross-contamination between liquid nitrogen and embryos may occur, and thus, the use of closed devices has been recommended to avoid the risk of contamination...
April 2018: Cryobiology
Bettina Krueger, Limin Yang, Christoph Korbmacher, Robert Rauh
The epithelial Na+ channel (ENaC) is a heteromeric channel composed of three subunits (α, β, γ). At the C-terminus of each subunit, a PY-motif allows binding of the ubiquitin ligase Nedd4-2 which plays a key role in promoting ENaC retrieval from the plasma membrane. Phosphorylation of Nedd4-2 by the serum and glucocorticoid-inducible kinase 1 (Sgk1) reduces Nedd4-2 binding to the PY-motifs. In β and γENaC, threonine residues (βT613, γT623) belong to an extracellular signal-regulated kinase (ERK) motif and directly precede the PY-motifs...
April 2018: Pflügers Archiv: European Journal of Physiology
Ronald H F Hunter, Fernando López-Gatius
Bearing in mind specific parallels between cow and human ovarian physiology, as noted in the manuscript, we have measured whether the temperature in a pre-ovulatory follicle is cooler than that in adjacent tissues. Using a novel approach not requiring anaesthetics or surgical procedures, we found that follicular fluid bathing cow oocytes shortly before ovulation is cooler than the neighbouring uterine surface and cooler than deep rectal temperature (the reference body temperature in cattle). By contrast, Graafian follicles of comparable size and ultrasonic image that do not subsequently ovulate do not have a reduced antral temperature...
January 29, 2018: Journal of Assisted Reproduction and Genetics
Steven Molinarolo, Sora Lee, Lilia Leisle, John D Lueck, Daniele Granata, Vincenzo Carnevale, Christopher A Ahern
Voltage-gated, sodium ion-selective channels (NaV ) generate electrical signals contributing to the upstroke of the action potential in animals. NaV s are also found in bacteria and are members of a larger family of tetrameric voltage-gated channels that includes CaV s, KV s, and NaV s. Prokaryotic NaV s likely emerged from a homotetrameric Ca2+ -selective voltage-gated progenerator, and later developed Na+ selectivity independently. The NaV signaling complex in eukaryotes contains auxiliary proteins, termed beta (β) subunits, which are potent modulators of the expression profiles and voltage-gated properties of the NaV pore, but it is unknown whether they can functionally interact with prokaryotic NaV channels...
April 6, 2018: Journal of Biological Chemistry
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