Ca2+-linked upregulation and mitochondrial production of nitric oxide in the mouse preimplantation embryo

Manser, Rosemary C.; Houghton, Franchesca D.

doi:10.1242/jcs.02927

Cited by 17 publications

(13 citation statements)

References 63 publications

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“…Measurement of mitochondrial activity by fluorescence was performed as previously described. 77 Embryos were incubated in KSOM supplemented with 500 nM MitoTracker ® Orange CMTMROS (Thermo Fisher Scientific, M7510) for 10 min, then washed three times in KSOM. Embryos were fixed in 4% paraformaldehyde for 30 min and were either mounted on glass slides for observation under a confocal microscope, or used for additional immunostaining.…”

Section: Methodsmentioning

confidence: 99%

The cytotoxic effects of dimethyl sulfoxide in mouse preimplantation embryos: a mechanistic study

et al. 2017

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Rationale: Dimethyl sulfoxide (DMSO) is commonly used as a solvent for water-insoluble substances, a vehicle for drug therapy, and a cryoprotectant for cultured cells. DMSO induced embryonic defects and its mechanism of action remains unclear. The rationale is based on the assumption that DMSO supplementation should induce long-term negative effects on both pre- and post-implantation embryo development.Methods: DMSO induced oxidative stress, ER stress, autophagy, mitophagy, signaling responsible genes and proteins were determined by RT-qPCR, Western blotting, immunofluorescence, and confocal microscopy. DMSO induced mitochondrial dysfunction was measured by transmission electron microcopy and JC-1 assay. Apoptosis was estimated using TUNEL and comet assay. Post-implantation embryo developmental capability was estimated by implantation site and fetus numbers.Results: Exposure to DMSO induced an early oxidative stress response within 0.5 to 2 h in 1-cell zygotes by disrupting the balance of pro- and anti-oxidants. Notably, DMSO-treated 2-cell embryos showed increased expression of unfolded protein response genes such as Hspa5, Hsp90b1, Ddit3, Atf4, and Xbp1. As a result, the development of many embryos is arrested at the 2-cell, 4-cell, or morula stages in a dose-dependent manner. Further, DMSO-induced endoplasmic reticulum stress increased mitochondrial Ca2+ levels, induced mitochondrial depolarization/dysfunction, and induced apoptotic cell death via the JNK/ATF2-dependent pathway. Consequently, treatment with DMSO increased the expression of autophagy initiation-, phagophore elongation-, and autophagosome formation-related genes, as well as localization of PINK1/Parkin, which are the main mediators of mitophagy, in mitochondria. Interestingly, DMSO causes cytotoxic effects in preimplantation embryos by inducing extensive mitophagy and autophagy. Especially, DMSO treatment decreased the inner cell mass and trophectoderm cell numbers as well as mRNA expression of B3gnt5 and Wnt3a in developed blastocysts, which decreased the implantation and developmental rates of full-term offspring after being transferred into pseudopregnant mice.Conclusion: These results provide a significant contribution to finding effective protective agents to combat DMSO mediated reproductive toxicity for application in human embryos in the near future.

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Section: Methodsmentioning

confidence: 99%

The cytotoxic effects of dimethyl sulfoxide in mouse preimplantation embryos: a mechanistic study

et al. 2017

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“…Recently, NO production has been documented in a variety of cell lines using fluorescent indicators (Manser andHoughton, 2006, Vatsa, et al, 2006). DAR-4M AM is a sensitive, cell-permeable NO fluorescent indicator with excitation maximum at ≈560 nm and emission maximum at ≈575 nm and high photo-stability, which make it suitable for live-cell imaging.…”

Section: Specificity Of Dar-4m Am For the Detection Of Rnsmentioning

confidence: 99%

Plasmodium falciparum: Food vacuole localization of nitric oxide-derived species in intraerythrocytic stages of the malaria parasite

Ostera

Tokumasu

Oliveira

et al. 2008

Experimental Parasitology

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Nitric oxide (NO) has diverse biological functions. Numerous studies have documented NO's biosynthetic pathway in a wide variety of organisms. Little is known, however, about NO production in intraerythrocytic Plasmodium falciparum. Using diaminorhodamine-4-methyl acetoxymethylester (DAR-4M AM), a fluorescent indicator, we obtained direct evidence of NO and NO-derived reactive nitrogen species (RNS) production in intraerythrocytic P. falciparum parasites, as well as in isolated food vacuoles from trophozoite stage parasites. We preliminarily identified two gene sequences that might be implicated in NO synthesis in intraerythrocytic P. falciparum. We showed localization of the protein product of one of these two genes, a molecule that is structurally similar to a plant nitrate reductase, in trophozoite food vacuole membranes. We confirmed previous reports on the antiproliferative effect of NOS (nitric oxide synthase) inhibitors in P.falciparum cultures; however, we did not obtain evidence that NOS inhibitors had the ability to inhibit RNS production or that there is an active NOS in mature forms of the parasite. We concluded that a nitrate reductase activity produce NO and NO-derived RNS in or around the food vacuole in P. falciparum parasites. The food vacuole is a critical parasitic compartment involved in hemoglobin degradation, heme detoxification and a target for antimalarial drug action. Characterization of this relatively unexplored synthetic activity could provide important clues into poorly understood metabolic processes of the malaria parasite,

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“…L-arginine is depleted from porcine embryo culture medium in vitro , suggesting that it is metabolized during embryonic development (Humpherson et al, 2005). Manser and Houghton (2006) utilized the nitric oxide sensitve probe DAF-FM diacetate and found that NO was present at all stages of murine preimplantation development. Supplementation of culture media with additional arginine improves porcine embryo development as well, suggesting that this amino acid plays a critical role in preimplantation development (Bauer et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

Arginine increases development of in vitro-produced porcine embryos and affects the protein arginine methyltransferase–dimethylarginine dimethylaminohydrolase–nitric oxide axis

Redel¹,

Tessanne²,

Spate³

et al. 2015

Reprod. Fertil. Dev.

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Culture systems promote development at rates lower than the in vivo environment. Here, we evaluated the embryo’s transcriptome to determine what the embryo needs during development. A previous mRNA sequencing endeavor found SLC7A1, an arginine transporter, to be up-regulated in in vitro compared to in vivo cultured embryos. We added different concentrations of arginine to our culture medium to meet the needs of the embryo. Increasing arginine from 0.12 to 1.69 mM improved the number of embryos that developed to the blastocyst stage. These blastocysts also had more total nuclei compared to controls and specifically, more trophectoderm nuclei. Embryos cultured in 1.69 mM arginine had lower SLC7A1 levels and had higher abundance of messages involved with glycolysis (HK1, HK2, and GPT2) and decreased expression of genes involved with blocking the TCA cycle (PDK1) as well as the pentose phosphate pathway (TALDO1). PRMT1, PRMT3 and PRMT5 expression throughout development was not affected by arginine. DDAH1 and DDAH2 message was found to be differentially regulated through development and DDAH2 protein was localized to the nuclei of blastocysts. Arginine has a positive effect on preimplantation development and may be affecting the NO-DDAH-PRMT axis.

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Ca2+-linked upregulation and mitochondrial production of nitric oxide in the mouse preimplantation embryo

Cited by 17 publications

References 63 publications

The cytotoxic effects of dimethyl sulfoxide in mouse preimplantation embryos: a mechanistic study

The cytotoxic effects of dimethyl sulfoxide in mouse preimplantation embryos: a mechanistic study

Plasmodium falciparum: Food vacuole localization of nitric oxide-derived species in intraerythrocytic stages of the malaria parasite

Arginine increases development of in vitro-produced porcine embryos and affects the protein arginine methyltransferase–dimethylarginine dimethylaminohydrolase–nitric oxide axis

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