bims-gamemb 2021-12-19 papers

bims-gamemb

Biomed News

on Gamete and embryo metabolism

Issue of 2021‒12‒19
ten papers selected by
Cameron A. Schmidt
East Carolina University

Sorbitol accumulation decreases oocyte quality in aged mice by altering the intracellular redox balance.
ROMO1 is required for mitochondrial metabolism during preimplantation embryo development in pigs.
LRRK2 regulates actin assembly for spindle migration and mitochondrial function in mouse oocyte meiosis.
The Chemical Environment at Maturation Stage in Pinus spp. Somatic Embryogenesis: Implications in the Polyamine Profile of Somatic Embryos and Morphological Characteristics of the Developed Plantlets.
Obesity and oocyte quality: Significant implications for ART and Emerging mechanistic insights.
Reactive oxygen species and their consequences on the structure and function of mammalian spermatozoa.
Dysregulated metabolism underpins Zika-virus-infection-associated impairment in fetal development.
Steroid levels in frog eggs: Manipulations, developmental changes, and implications for maternal steroid effects.
Integrative proteomics and phosphoproteomics reveals phosphorylation networks involved in the maintenance and expression of embryogenic competence in sugarcane callus.
Capillary electrophoresis - mass spectrometry as a tool for the non-invasive target metabolomic analysis of underivatized amino acids for evaluating embryo viability in assisted reproduction.

Aging (Albany NY). 2021 Dec 12. 13(undefined):

Sorbitol accumulation decreases oocyte quality in aged mice by altering the intracellular redox balance.

Yuexin Zhang, Zhengjie Yan, Hanwen Liu, Lingjun Li, Chun Yuan, Lianju Qin, Lingbo Cai, Jiayin Liu, Yanqiu Hu, Yugui Cui.

  Sorbitol is a product of glucose metabolism through the polyol pathway. Many studies have demonstrated that excessive sorbitol can disrupt the intracellular redox balance. However, we still know very little about the impact of excessive intracellular sorbitol on oocyte quality, oocyte maturation, and embryo developmental potential. This study explored whether intracellular sorbitol accumulates in the oocytes of aged mice during in vitro maturation (IVM) and what roles sorbitol plays in oocyte development and maturation. Our results showed that sorbitol levels were significantly higher in in vitro-matured oocytes from aged mice than in oocytes from young mice (14.08 ± 3.78 vs. 0.23 ± 0.04 ng/oocyte). The expression of aldose reductase (AR) mRNA was significantly higher in the in vitro-cultured oocytes from 9-month-old mice than prior to culture. To decrease the excessive intracellular sorbitol in oocytes from aged mice, sorbinil, a specific inhibitor of aldose reductase, was supplemented in IVM medium, and the sorbitol level was significantly decreased (14.08 ± 3.78 vs. 0.48 ± 0.19 ng/oocyte). Our results indicated that the percentage of oocytes with first polar body extrusion (PBE) was significantly higher in the sorbinil group than in the aged group (82.4% ± 7.2% vs. 66.1% ± 6.9%), and the content of sorbitol was drastically increased in the aged group. The ROS fluorescence intensity in the sorbinil group was drastically lower than that in the aged group, while the GSH fluorescence intensity was significantly higher. Interestingly, SOD1 was upregulated in the sorbinil group. The present study suggests that excessive sorbitol accumulation is induced during IVM in aged mouse oocytes, which negatively influences oocyte quality by altering the intracellular redox balance. Inhibition of sorbitol accumulation may be a potential method to improve the nuclear maturation of aged oocytes.

Keywords:  aged mouse oocytes; in vitro maturation of oocytes; reactive oxygen species; sorbinil; sorbitol

DOI:  https://doi.org/10.18632/aging.203747
Cell Div. 2021 Dec 16. 16(1): 7

ROMO1 is required for mitochondrial metabolism during preimplantation embryo development in pigs.

Dongjie Zhou, Ming-Hong Sun, Song-Hee Lee, Xiang-Shun Cui.

  BACKGROUND: Reactive oxygen species (ROS) modulator 1 (ROMO1) is a mitochondrial membrane protein that is essential for the regulation of mitochondrial ROS production and redox sensing. ROMO1 regulates ROS generation within cells and is involved in cellular processes, such as cell proliferation, senescence, and death. Our purpose is to investigates the impact of ROMO1 on the mitochondria during porcine embryogenesis.RESULTS: We found that high expression of ROMO1 was associated with porcine preimplantation embryo development, indicating that ROMO1 may contribute to the progression of embryogenesis. Knockdown of ROMO1 disrupted porcine embryo development and blastocyst quality, thereby inducing ROS production and decreasing mitochondrial membrane potential. Knockdown of ROMO1 induced mitochondrial dysfunction by disrupting the balance of OPA1 isoforms to release cytochrome c, reduce ATP, and induce apoptosis. Meanwhile, ROMO1 overexpression showed similar effects as ROMO1 KD on the embryos. Overexpression of ROMO1 rescued the ROMO1 KD-induced defects in embryo development, mitochondrial fragmentation, and apoptosis.
CONCLUSIONS: ROMO1 plays a critical role in embryo development by regulating mitochondrial morphology, function, and apoptosis in pigs.

Keywords:  Apoptosis; Embryo development; Mitochondria; Porcine; ROMO1

DOI:  https://doi.org/10.1186/s13008-021-00076-7
J Mol Cell Biol. 2021 Dec 16. pii: mjab079. [Epub ahead of print]

LRRK2 regulates actin assembly for spindle migration and mitochondrial function in mouse oocyte meiosis.

Zhen-Nan Pan, Jing-Cai Liu, Jia-Qian Ju, Yue Wang, Shao-Chen Sun.

  LRRK2 belongs to the Roco GTPase family and is a large multi-domain protein harboring both GTPase and kinase activities. LRRK2 plays indispensable roles in many processes, such as autophagy and vesicle trafficking in mitosis. In the present study, we showed the critical roles of LRRK2 in mammalian oocyte meiosis. LRRK2 is mainly accumulated at the meiotic spindle periphery during oocyte maturation. Depleting LRRK2 led to the polar body extrusion defects and also induced large polar bodies in mouse oocytes. Mass spectrometry analysis and co-immunoprecipitation results showed that LRRK2 was associated with several actin-regulating factors such as Fascin and ROCK, and depletion of LRRK2 affected the expression of ROCK, phosphorylated cofilin, and Fascin. Further analysis showed that LRRK2 depletion did not affect spindle organization but caused the failure of spindle migration, which was largely due to the decrease of cytoplasmic actin filaments. Moreover, LRRK2 showed similar localization pattern with mitochondria, and LRRK2 was associated with several mitochondria-related proteins. Indeed, mitochondrial distribution and function were all disrupted in LRRK2-depleted oocytes. In summary, our results indicated the critical roles of LRRK2 in actin assembly for spindle migration and mitochondrial function in mouse oocyte meiosis.

Keywords:  LRRK2; actin; meiosis; oocyte; spindle

DOI:  https://doi.org/10.1093/jmcb/mjab079
Front Plant Sci. 2021 ;12 771464

The Chemical Environment at Maturation Stage in Pinus spp. Somatic Embryogenesis: Implications in the Polyamine Profile of Somatic Embryos and Morphological Characteristics of the Developed Plantlets.

Antonia Maiara Marques do Nascimento, Luiza Giacomolli Polesi, Franklin Panato Back, Neusa Steiner, Miguel Pedro Guerra, Ander Castander-Olarieta, Paloma Moncaleán, Itziar Aurora Montalbán.

  Changes in the chemical environment at the maturation stage in Pinus spp. somatic embryogenesis will be a determinant factor in the conversion of somatic embryos to plantlets. Furthermore, the study of biochemical and morphological aspects of the somatic embryos could enable the improvement of somatic embryogenesis in Pinus spp. In the present work, the influence of different amino acid combinations, carbohydrate sources, and concentrations at the maturation stage of Pinus radiata D. Don and Pinus halepensis Mill. was analyzed. In P. radiata, the maturation medium supplemented with 175 mM of sucrose and an increase in the amino acid mixture (1,100 mgL-1 of L-glutamine, 1,050 mgL-1 of L-asparagine, 350 mgL-1 of L-arginine, and 35 mgL-1 of L-proline) promoted bigger embryos, with a larger stem diameter and an increase in the number of roots in the germinated somatic embryos, improving the acclimatization success of this species. In P. halepensis, the maturation medium supplemented with 175 mM of maltose improved the germination of somatic embryos. The increase in the amount of amino acids in the maturation medium increased the levels of putrescine in the germinated somatic embryos of P. halepensis. We detected significant differences in the amounts of polyamines between somatic plantlets of P. radiata and P. halepensis; putrescine was less abundant in both species. For the first time, in P. radiata and P. halepensis somatic embryogenesis, we detected the presence of cadaverine, and its concentration changed according to the species.

Keywords:  Pinus halepensis; Pinus radiata; amino acids; osmolality; sugars

DOI:  https://doi.org/10.3389/fpls.2021.771464
Biol Reprod. 2021 Dec 17. pii: ioab228. [Epub ahead of print]

Obesity and oocyte quality: Significant implications for ART and Emerging mechanistic insights.

Macarena B Gonzalez, Rebecca L Robker, Ryan D Rose.

  The prevalence of obesity in adults worldwide, and specifically in women of reproductive age, is concerning given the risks to fertility posed by the increased risk of type 2 diabetes, metabolic syndrome and other non-communicable diseases. Obesity has a multi-systemic impact in female physiology that is characterized by the presence of oxidative stress, lipotoxicity, and the activation of pro-inflammatory pathways, inducing tissue-specific insulin resistance and ultimately conducive to abnormal ovarian function. A higher body mass is linked to Polycystic Ovary Syndrome, dysregulated menstrual cycles, anovulation, and longer time to pregnancy, even in ovulatory women. In the context of ART, compared to women of normal BMI, obese women have worse outcomes in every step of their journey, resulting in reduced success measured as live birth rate. Even after pregnancy is achieved, obese women have a higher chance of miscarriage, gestational diabetes, pregnancy complications, birth defects, and most worryingly, a higher risk of stillbirth and neonatal death. The potential for compounding effects of ART on pregnancy complications and infant morbidities in obese women has not been studied. There is still much debate in the field on whether these poorer outcomes are mainly driven by defects in oocyte quality, abnormal embryo development or an unaccommodating uterine environment, however the clinical evidence to date suggests a combination of all three are responsible. Animal models of maternal obesity shed light on the mechanisms underlaying the effects of obesity on the peri-conception environment, with recent findings pointing to lipotoxicity in the ovarian environment as a key driver of defects in oocytes that have not only reduced developmental competence but long-lasting effects in offspring health.

Keywords:  ART; IVF/ICSI; obesity; oocyte; oocyte quality; reproductive success; women’s health

DOI:  https://doi.org/10.1093/biolre/ioab228
Antioxid Redox Signal. 2021 Dec 16.

Reactive oxygen species and their consequences on the structure and function of mammalian spermatozoa.

Joel R Drevet, Jorge Hallak, Mohammad Hossein Nasr-Esfahani, Robert John Aitken.

Among the two hundred or so cell types that make up mammals, spermatozoa have an ambiguous relationship with the reactive oxygen species (ROS) inherent in the consumption of oxygen that supports aerobic metabolism. In this review, we shall see that on the one hand, spermatozoa need the action of ROS to reach their structural and functional maturity. On the other hand, we shall also see that due to intrinsic characteristics that make these cells unique, they are, perhaps more than any other cell type, susceptible to oxidative damage. Oxidative stress can have dramatic consequences for the spermatozoon itself, but also, and above all, on its primary objective, which is to carry out fertilization and to ensure, in part, that the embryonic development program should lead to a healthy progeny. Recent studies, mainly in animal models but also through clinical investigations, have improved our knowledge of how oxidative damage affects sperm structures and functions. The focus of this review will be on how genetic and epigenetic oxidative alterations to spermatozoa can have dramatic unintended consequences in terms of both the support and the suppression of sperm function.

DOI: https://doi.org/10.1089/ars.2021.0235
Cell Rep. 2021 Dec 14. pii: S2211-1247(21)01612-0. [Epub ahead of print]37(11): 110118

Dysregulated metabolism underpins Zika-virus-infection-associated impairment in fetal development.

Clement Yau, John Z H Low, Esther S Gan, Swee Sen Kwek, Liang Cui, Hwee Cheng Tan, Darren Z L Mok, Candice Y Y Chan, October M Sessions, Satoru Watanabe, Subhash G Vasudevan, Yie Hou Lee, Kuan Rong Chan, Eng Eong Ooi.

  Zika virus (ZIKV) is an Aedes-mosquito-borne flavivirus that causes debilitating congenital and developmental disorders. Improved understanding of ZIKV pathogenesis could assist efforts to fill the therapeutic and vaccine gap. We use several ZIKV strains, including a pair differing by a single phenylalanine-to-leucine substitution (M-F37L) in the membrane (M) protein, coupled with unbiased genomics to demarcate the border between attenuated and pathogenic infection. We identify infection-induced metabolic dysregulation as a minimal set of host alterations that differentiates attenuated from pathogenic ZIKV strains. Glycolytic rewiring results in impaired oxidative phosphorylation and mitochondrial dysfunction that trigger inflammation and apoptosis in pathogenic but not attenuated ZIKV strains. Critically, pyruvate supplementation prevents cell death, in vitro, and rescues fetal development in ZIKV-infected dams. Our findings thus demonstrate dysregulated metabolism as an underpinning of ZIKV pathogenicity and raise the potential of pyruvate supplementation in expectant women as a prophylaxis against congenital Zika syndrome.

Keywords:  Zika virus; attenuated; congenital zika syndrome; glycolysis; pathogenic; pyruvate supplementation; tricarboxylic acid cycle

DOI:  https://doi.org/10.1016/j.celrep.2021.110118
J Exp Zool A Ecol Integr Physiol. 2021 Dec 14.

Steroid levels in frog eggs: Manipulations, developmental changes, and implications for maternal steroid effects.

Ryan T Paitz, Matthew B Dugas.

  Exposure to maternally derived steroids during embryonic development can elicit phenotypic effects in the resulting offspring. Studies of maternal steroid effects, especially rich in mammals and birds, have offered exciting insights into the evolution of maternal effects in vertebrates. To extend this literature, we quantified levels of steroids in the eggs of four neotropical dendrobatid frogs that lay terrestrial clutches, a reproductive strategy that has evolved multiple times in amphibians. Building on our observational results, we then manipulated levels of pregnenolone and progesterone in eggs of one species and examined how this affected steroid levels during development. Eggs of all four species had detectable steroids levels, with progestogens being more abundant than androgens and glucocorticoids. Estrogens could not be detected. Immersion of frog eggs in a solution containing pregnenolone and progesterone resulted in elevated levels of both steroids early in development, but levels declined and were similar to those in unmanipulated eggs by the end of development. Treated eggs also exhibited a transient increase in levels of steroids that can be produced from pregnenolone and progesterone. Overall, our findings demonstrate that frog eggs contain steroids similar to what has been observed in other egg-laying vertebrates. During development, steroid levels are dynamic, further suggesting developing embryos regulate exposure to maternal steroids. These results set the stage for investigating the causes and consequences of maternal steroid effects in frogs.

Keywords:  anura; developmental endocrinology; maternal effects; poison frogs

DOI:  https://doi.org/10.1002/jez.2566
J Plant Physiol. 2021 Dec 08. pii: S0176-1617(21)00226-1. [Epub ahead of print]268 153587

Integrative proteomics and phosphoproteomics reveals phosphorylation networks involved in the maintenance and expression of embryogenic competence in sugarcane callus.

Lucas R Xavier, Felipe A Almeida, Vitor B Pinto, Lucas Z Passamani, Claudete Santa-Catarina, Gonçalo A de Souza Filho, Brian P Mooney, Jay J Thelen, Vanildo Silveira.

  Plant embryogenic cell culture allows mass propagation and genetic manipulation, but the mechanisms that determine the fate of these totipotent cells in somatic embryos have not yet been elucidated. Here, we performed label-free quantitative proteomics and phosphoproteomics analyses to determine signaling events related to sugarcane somatic embryo differentiation, especially those related to protein phosphorylation. Embryogenic calli were compared at multiplication (EC0, dedifferentiated cells) and after 14 days of maturation (EC14, onset of embryo differentiation). Metabolic pathway analysis showed enriched lysine degradation and starch/sucrose metabolism proteins during multiplication, whereas the differentiation of somatic embryos was found to involve the enrichment of energy metabolism, including the TCA cycle and oxidative phosphorylation. Multiplication-related phosphoproteins were associated with transcriptional regulation, including SNF1 kinase homolog 10 (KIN10), SEUSS (SEU), and LEUNIG_HOMOLOG (LUH). The regulation of multiple light harvesting complex photosystem II proteins and phytochrome interacting factor 3-LIKE 5 were predicted to promote bioenergetic metabolism and carbon fixation during the maturation stage. A motif analysis revealed 15 phosphorylation motifs. The [D-pS/T-x-D] motif was overrepresented during somatic embryo differentiation. A protein-protein network analysis predicted interactions among SNF1-related protein kinase 2 (SnRK2), abscisic acid-responsive element-binding factor 2 (ABF2), and KIN10, which indicated the role of these proteins in embryogenic competence. The predicted interactions between TOPLESS (TPL) and histone deacetylase 19 (HD19) may be involved in posttranslational protein regulation during somatic embryo differentiation. These results reveal the protein regulation dynamics of somatic embryogenesis and new players in somatic embryo differentiation, including their predicted phosphorylation motifs and phosphosites.

Keywords:  Embryo development; Label-free phosphoproteomies; Somatic Embryogenesis; Sugarcane

DOI:  https://doi.org/10.1016/j.jplph.2021.153587
Electrophoresis. 2021 Dec 11.

Capillary electrophoresis - mass spectrometry as a tool for the non-invasive target metabolomic analysis of underivatized amino acids for evaluating embryo viability in assisted reproduction.

Monika Skrutková Langmajerová, Marta Pelcová, Pavla Vedrová, Andrea Celá, Zdeněk Glatz.

  Monitoring metabolite uptake and excretion in the culture medium is a non-invasive technique that is used for the metabolic study of cleaving embryos after in vitro fertilization. Low sample consumption, the versatility of the detection and optimal sensitivity and selectivity are essential elements for extracellular metabolome analyses, and can be conveniently achieved by combining capillary electrophoresis with mass spectrometric detection. This paper reports a method for amino acid determination in a limited volume sample (8μL) of spent culture media collected after the cultivation of in vitro fertilized embryos. Special attention was focused on the sample preparation procedure. The sample was processed with acetonitrile, which facilitates online sample preconcentration via field-amplified sample stacking, and undesired sample evaporation was significantly reduced by the simultaneous addition of dimethyl sulfoxide. Key parameters that affected electrophoretic separation and mass spectrometric detection were investigated, including the type of buffers and organic solvent, optimization of their concentrations, and finally the settings for their ionization. The separation and quantification of 19 amino acids were achieved using 15% HAc as the background electrolyte with a sheath liquid consisting of an equimolar mixture of methanol and water. The applicability of the optimized system was demonstrated by determining the amino acid profile in 40 samples of spent cultivation medium in this pilot study. This developed method also has great potential for amino acid analyses in minute sample volumes of other biological matrices. This article is protected by copyright. All rights reserved.

Keywords:  Amino acids; Capillary electrophoresis; Culture media; Embryo metabolism; Mass spectrometry

DOI:  https://doi.org/10.1002/elps.202100328