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Molecular Reproduction and Development

Zer Vue, Gabriel Gonzalez, C Allison Stewart, Shyamin Mehra, Richard R Behringer
Endometrial or uterine glands secrete substances essential for uterine receptivity to the embryo, implantation, conceptus survival, and growth. Adenogenesis is the process of gland formation within the stroma of the uterus. In the mouse, uterine gland formation initiates at postnatal day (P) 5. Uterine gland morphology is poorly understood because it is primarily based on two-dimensional (2D) histology. To more fully describe uterine gland morphogenesis, we generated three-dimensional (3D) models of postnatal uterine glands from P0 to P21, based on volumetric imaging using light sheet microscopy...
March 15, 2018: Molecular Reproduction and Development
Laura Masala, Federica Ariu, Luisa Bogliolo, Emanuela Bellu, Sergio Ledda, Daniela Bebbere
Oocytes from prepubertal animals have a reduced ability to undergo embryo development and produce viable offspring. The present work used an ovine model consisting of oocytes derived from adult and prepubertal donors to assess the molecular status of oocytes and preimplantation embryos with different developmental competence. The lower potential of oocytes of young donors was confirmed in terms of in vitro developmental capabilities and kinetics. A panel of genes including maternal effect (DPPA3, GDF9, NMP2, ZAR1) and housekeeping genes (ACTB, RPL19, SDHA, YWHAZ, ATP1A1), genes involved in DNA methylation (DNMT1, DNMT3A, DNMT3B), genomic imprinting (IGF2R), pluripotency (NANOG, POU5F1) and cell cycle regulation (CCNB1, CDK1, MELK) was relatively quantified...
March 15, 2018: Molecular Reproduction and Development
Francesca Mossa, Daniela Bebbere, Antonello Ledda, Giovanni P Burrai, Imane Chebli, Elisabetta Antuofermo, Sergio Ledda, Antonello Cannas, Francesco Fancello, Alberto S Atzori
Maternal nutrition during critical gestation periods impacts on offspring in later life; effects of high-starch maternal diet on testicular development in lambs were addressed. Dairy ewes were fed diets providing either 27% (Starch, S) or 11% (Fiber, F) of starch from mating to lambing (∼147 days; S147, F147) or for the last 75 days of gestation (S75, F75). Testes of single male lambs were measured and then sampled for histological and gene expression analyses at selected ages. Testicular dimensions and weight were similar among groups, but the total area of seminiferous tubules increased with age and tended to be higher (P = 0...
March 15, 2018: Molecular Reproduction and Development
Verónica M Negrón-Pérez, Luana Teixeira Rodrigues, Gisele Zoccal Mingoti, Peter J Hansen
Rho-associated coiled-coil containing protein kinases (ROCK1 and ROCK2) are activated by binding to RHO GTPases and phosphorylate a variety of downstream targets including actinomyosin. In the mouse embryo, ROCK signaling acts to promote formation of trophectoderm (TE) and inhibit formation of the inner cell mass (ICM) by polarizing outer cells of the embryo to inactivate Hippo signaling (Kono et al., 2014; Mihajlović and Bruce, 2016). This article is protected by copyright. All rights reserved.
March 15, 2018: Molecular Reproduction and Development
Veronika L Flöter, Anne-Kathrin Lorenz, Benedikt Kirchner, Michael W Pfaffl, Stefan Bauersachs, Susanne E Ulbrich
Porcine conceptuses synthesize estrogens between day 11 and 12 as signal for maternal recognition of pregnancy. A preimplantational estrogen exposure to pregnant gilts has been associated with embryonic losses and changes in endometrial mRNA expression. MicroRNAs (miRNAs) play a key role in the mRNA regulation by modulating the expression. Effects of estrogens on endometrial miRNAs have not been investigated in this context so far. Thus, we studied the endometrial expression profile of miRNAs in the pig at gestational day 10 after daily estradiol-17β (E2) application starting at fertilization using either 0, 0...
March 15, 2018: Molecular Reproduction and Development
G Charles Ostermeier, Cristina Cardona, Melissa A Moody, Alana J Simpson, Romeo Mendoza, Eric Seaman, Alexander J Travis
Sperm must mature functionally in the process of capacitation to become able to fertilize. Capacitation depends on membrane lipid changes, and can be quantitatively assessed by redistribution of the ganglioside GM1 , the basis of the Cap-Score™ sperm function test. Here, differences in Cap-Score were compared among and within men at two time points. Ejaculates were liquefied, washed, and incubated for 3 hours under capacitating (Cap) conditions, then fixed and analyzed immediately (Day0); after being incubated 3 hours under Cap conditions then maintained 22-24 hours in fix (Day1-fix); or after 22-24 hours incubation under Cap conditions prior to fixation (Day1)...
March 9, 2018: Molecular Reproduction and Development
Narumi Ogonuki, Hiroki Inoue, Shogo Matoba, Yoko Kakino Kurotaki, Hidetoshi Kassai, Yukiko Abe, Erika Sasaki, Atsu Aiba, Atsuo Ogura
The common marmoset (Callithrix jacchus) represents a promising nonhuman primate model for the study of human diseases because of its small size, ease of handling, and availability of gene-modified animals. Here, we aimed to devise reproductive technology for marmoset spermatid injection using immature males for a possible rapid generational turnover. Spermatids at each step could be identified easily by their morphology under differential interference microscopy: thus, early round spermatids had a round nucleus with a few nucleolus-like structures and abundant cytoplasm, as in other mammals...
February 19, 2018: Molecular Reproduction and Development
V Duranthon, P Chavatte-Palmer
Early stages of mammalian embryonic development are now known to be very sensitive to their microenvironment, with long term effects on fetal, postnatal and adult health, thus extending to these early stages the concept of Developmental Origin of Health and Disease (DoHaD). In this scientific context, and with 3% of births in developed countries, safety of Assisted Reproductive Techniques procedures becomes a matter of concern. Besides, embryo technologies in domestic mammals, using huge number of embryos, do not seem to evidence heavy impacts on adult phenotypes...
February 19, 2018: Molecular Reproduction and Development
Seok Hee Lee, Hyun Ju Oh, Min Jung Kim, Erif Maha Nugraha Setyawan, Yoo Bin Choi, Byeong Chun Lee
Human endothelial progenitor cells (EPCs) have been applied to regenerative medicine for their roles in angiogenesis as well as neovascularization, and these angiogenetic functions have beneficial effects on maturation of ovarian follicles. However, little information is available on whether EPCs on culture systems affect oocyte maturation and subsequent embryo development. Therefore, the objective of this study was to investigate the effect of EPC co-culture on porcine oocytes during in vitro maturation (IVM) and subsequent embryo development, and to examine gene expression in cumulus cells, oocytes and blastocysts...
February 14, 2018: Molecular Reproduction and Development
Monica A Stoops, G Douglas Winget, Christopher J DeChant, Ray L Ball, Terri L Roth
Genetic sexing of animals with long gestation time benefits the management of captive populations. Here, X and Y chromosome-specific primers, based on equine gene sequencing data, were developed and tested on captive rhinoceroses (10 males, 20 females) representing four species (Diceros bicornis, Certaotherium simum simum, Rhinoceros unicornis, and Dicerorhinus sumatrensis). The Y chromosome-specific primer set targeted SRY (Sex-determining region Y), and amplified a 177-bp product following PCR of DNA extracted from males, but not females, of all species...
February 13, 2018: Molecular Reproduction and Development
Chong Li, Shiwei Xiao, Jie Hao, Xiaogang Liao, Gang Li
Cryptochrome (Cry)1 is essential for generating circadian rhythm in central and many peripheral oscillators; however, its role in male reproduction remains unclear. We investigated this question using Cry1 knockout (KO) mice. We found that Cry1 is necessary for normal testicular function: Cry1 deficiency increased testicular germ cell apoptosis and decreased sperm count. A transcriptome analysis showed that the expression levels of 375 genes-including 12 encoding micro (mi)RNAs-were altered in the testis of Cry1 KO mice relative to wild-type controls...
February 7, 2018: Molecular Reproduction and Development
Ashley L Severance, Keith E Latham
Oocytes uniquely accumulate cytoplasmic constituents to support early embryogenesis. This unique specialization is accompanied by acquisition of a large size and by execution of asymmetric meiotic divisions that preserve precious ooplasm through the expulsion of minimal size polar bodies. While often taken for granted, these basic features of oogenesis necessitate unique specializations of the meiotic apparatus. These include a chromatin-sourced RanGTP gradient that restricts spindle size by defining a spatial domain where meiotic spindles form, acentriolar centrosomes which relies on microtubule organizing centers to form spindle poles, and an actin-based mechanism for asymmetric spindle positioning...
February 7, 2018: Molecular Reproduction and Development
Md Shahanoor Alam, Hideaki Moriyama, Midori Matsumoto
The DUT gene encodes Deoxyuridine triphosphatase (dUTPase), which is involved in nucleotide metabolism. dUTPase prevents uracil misincorporation in DNA by balancing the intracellular ratio between dUTP and dTTP. This study aimed to investigate the role of Dr-dut gene in the planarian Dugesia ryukyuensis by assessing the consequences of Dr-dut silencing on known phenomena, including regeneration following amputation and radiation damage. We functionally disrupted planarian Dr-dut mRNA by feeding RNAi-containing food to animals...
February 5, 2018: Molecular Reproduction and Development
Lin Tang, Alla Bondareva, Raquel González, Jose Rafael Rodriguez-Sosa, Daniel F Carlson, Dennis Webster, Scott Fahrenkrug, Ina Dobrinski
Spermatogonia represent a diploid germ cell population that includes spermatogonial stem cells. In this report, we describe new methods for isolation of highly enriched porcine spermatogonia based on light scatter properties, and for targeted mutagenesis in porcine spermatogonia using nucleofection and TALENs. We optimized a nucleofection protocol to deliver TALENs specifically targeting the DMD locus in porcine spermatogonia. We also validated specific sorting of porcine spermatogonia based on light scatter properties...
February 2, 2018: Molecular Reproduction and Development
Wei Li, Amy S Miner, Ling Zhang, Paul Ratz, Zhibing Zhang
Mouse meiosis expressed gene 1 (Meig1) was originally cloned in a search for genes essential for meiosis. Early studies suggested that the protein forms a dimer and is phosphorylated in male germ cells. However, studies from our laboratory demonstrated that the gene is critical for the final phase, spermiogenesis. As a monomer, MEIG1 binds to PACRG through a specific domain on the protein surface. Here by using 2D Western blot analysis and the Pro-Q Diamond phosphoprotein gel stain, we show that mouse MEIG1 does not undergo major phosphorylation in the testis...
February 2, 2018: Molecular Reproduction and Development
Keiya Nagasawa, Kasane Imura-Kishi, Aya Uchida, Ryuji Hiramatsu, Masamichi Kurohmaru, Yoshiakira Kanai
In mouse testes, Sertoli cells support the continuous process of spermatogenesis, which is dependent on seminiferous epithelial cycles along the longitudinal axis of the seminiferous tubule. Sertoli cell function is modulated partly by local cytokines and/or growth factors derived from adjacent tissues such as blood vessels, macrophages, rete testis, etc. However, the spatial activation patterns by local signals in vivo remain unclear. In this study, we focused on Signal Transducers and Activators of Transcription (STAT) signalling in Sertoli cells, because STAT is a major crucial cytokine transducer for somatic cyst cell regulation in Drosophila testis niches...
February 2, 2018: Molecular Reproduction and Development
Ismail Kaarouch, Nouzha Bouamoud, Aicha Madkour, Noureddine Louanjli, Brahim Saadani, Said Assou, Smahane Aboulmaouahib, Saaid Amzazi, Henri Copin, Moncef Benkhalifa, Sefrioui Omar
This study assessed sperm quality declining on relation to paternal age and its impact on in vitro fertilization (IVF) outcomes in order to estimate the APA (Advanced Paternal Age) cutoff. For this, 83 couples undergoing IVF treatment for male factor infertility were enrolled. The women age was ≤39 years, whereas the men were divided in two groups: APA (n = 41; age≥40 years) and young (Y) (n = 42; age < 40 years). Conventional semen parameters (volume, concentration, motility, vitality and morphology) were analyzed in the collected sperm samples...
February 2, 2018: Molecular Reproduction and Development
B R Mordhorst, S L Murphy, R M Ross, J A Benne, M S Samuel, R F Cecil, B K Redel, L D Spate, C N Murphy, K D Wells, J A Green, R S Prather
Somatic cell nuclear transfer is a valuable technique for the generation of genetically engineered animals, however the efficiency of cloning in mammalian species is low (1-3%). Differentiated somatic cells commonly used in nuclear transfer utilize the tricarboxylic acid cycle and cellular respiration for energy production. Comparatively the metabolism of somatic cells contrasts that of the cells within the early embryos which predominately use glycolysis. Early embryos (prior to implantation) are evidenced to exhibit characteristics of a Warburg Effect (WE)-like metabolism...
February 2, 2018: Molecular Reproduction and Development
Sánchez-Santos A, Martínez-Hernández Mg, Contreras-Ramos A, Ortega-Camarillo C, Baiza-Gutman LA
During embryo implantation, the outer layer of the blastocyst interacts with the endometrium giving rise to the development of the trophoblast cell lineage. The cells in this lineage participate in the penetration of endometrium due to their motility and invasive properties. The mechanisms that regulate the differentiation and invasive ability of these cells are essential for the establishment and maintenance of an efficient exchange between maternal and fetal tissues during pregnancy. In this context, hyperglycemia can induce oxidative stress causing alterations in the placenta...
February 2, 2018: Molecular Reproduction and Development
Yasmin Schuermann, Dayananda Siddappa, Melissa Pansera, Raj Duggavathi
Successful ovulation requires the actions of gonadotropins along with those mediated by growth factors binding to their receptor tyrosine kinases (RTKs). There are several growth factors such as epidermal growth factor family ligands and interleukins that play a role during ovulation initiated by the preovulatory surge of luteinizing hormone (LH). The aim of this project was to analyze growth factor signaling pathways induced by LH in mouse granulosa cells. Immature female mice were treated with equine chorionic gonadotropin (eCG) followed 48h later by human chorionic gonadotropin (hCG) to induce follicular growth and ovulation...
February 2, 2018: Molecular Reproduction and Development
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