New technique to quantify the lipid composition of lipid droplets in porcine oocytes and pre-implantation embryos using Nile Red fluorescent probe

Romek, M.; Gajda, B.; Krzysztofowicz, Ewa; Kępczyński, Mariusz; Smorąg, Z.

doi:10.1016/j.theriogenology.2010.06.040

Cited by 57 publications

(33 citation statements)

References 42 publications

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“…Lipophilic Nile red dye is also commonly used to visualise lipid in oocytes; oocytes that have darker cytoplasm have greater Nile red staining (Leroy et al 2005b). Decomposition of Nile red spectra was used to quantify triacylglycerol, phospholipid and cholesterol in oocytes and embryos (Romek et al 2011); Nile Red staining of porcine oocytes followed by florescence resonance energy transfer was used to demonstrate that lipid droplets and mitochondria are within w10 nm of each other (Sturmey et al 2006). BODIPY 493/503 is a neutral lipid dye that has more recently been used to detect lipid droplets in oocytes and demonstrate they are bounded by the lipid droplet protein ADRP in both mouse and bovine , Aardema et al 2011.…”

Section: Lipid Droplets In Oocytesmentioning

confidence: 99%

Lipids and oocyte developmental competence: the role of fatty acids and β-oxidation

2014

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Metabolism and ATP levels within the oocyte and adjacent cumulus cells are associated with quality of oocyte and optimal development of a healthy embryo. Lipid metabolism provides a potent source of energy and its importance during oocyte maturation is being increasingly recognised. The triglyceride and fatty acid composition of ovarian follicular fluid has been characterised for many species and is influenced by nutritional status (i.e. dietary fat, fasting, obesity and season) as well as lactation in cows. Lipid in oocytes is a primarily triglyceride of specific fatty acids which differ by species, stored in distinct droplet organelles that re-localise during oocyte maturation. The presence of lipids, particularly saturated vs unsaturated fatty acids, in in vitro maturation systems affects oocyte lipid content as well as developmental competence. Triglycerides are metabolised by lipases that have been localised to cumulus cells as well as oocytes. Fatty acids generated by lipolysis are further metabolised by b-oxidation in mitochondria for the production of ATP. b-oxidation is induced in cumulus-oocyte complexes (COCs) by the LH surge, and pharmacological inhibition of b-oxidation impairs oocyte maturation and embryo development. Promoting b-oxidation with L-carnitine improves embryo development in many species. Thus, fatty acid metabolism in the mammalian COC is regulated by maternal physiological and in vitro environmental conditions; and is important for oocyte developmental competence.

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Section: Lipid Droplets In Oocytesmentioning

confidence: 99%

Lipids and oocyte developmental competence: the role of fatty acids and β-oxidation

2014

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“…We used Nile red staining and then assessed the digital images obtained by fluorescence microscopy to quantify the accumulation of neutral lipids (cholesterol, lipoproteins, and triglycerides) in intracellular fat vacuoles [14][15][16].…”

Section: Lipid Accumulation Quantificationmentioning

confidence: 99%

“…For example, oocytes may use intracellular triglycerides as an energy source and different types of phospholipids regulate oocyte maturation. The ratio of phospholipids to cholesterol also influences the physical properties of their biological membranes [16,34,35].…”

Section: The Effect Of Maternal Body Condition On Fertility and Lipidmentioning

confidence: 99%

The effect of maternal body condition on in vivo production of zygotes and behavior of delivered offspring in mice

et al. 2015

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“…After this period, the oocytes and embryos were washed 3 x in PBS + PVP and transferred to a glass slide (12 oocytes or embryos/slide) containing 5 μL Vectashield mounting medium (Brunschwig Chemie, Amsterdam, Netherlands), which was carefully covered with a coverslip. The fluorescence of Nile Red was evaluated under an epifluorescence microscope (Leica DM 4500B; excitation 515-560 nm and emission greater than 590 nm) with fast resolution [31]. The photographs of stained oocytes or embryos (20× magnification) were taken at 80 ms exposure and the fluorescence intensity (FI) was measured using Im-ageJ software.…”

Section: Determination Of Lipid Content With the Fluorescent Probe Nimentioning

confidence: 99%

Supplementing in vitro embryo production media by NPPC and sildenafil affect the cytoplasmic lipid content and gene expression of bovine cumulus-oocyte complexes and embryos

Botigelli

Razza

Pioltine

et al. 2018

Reproductive Biology

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In our study, we added natriuretic peptide type C (NPPC) and/or sildenafil during in vitro maturation (IVM) of bovine cumulus-oocyte complexes (COCs) followed by in vitro culture (IVC) of embryos with or without sildenafil. We evaluated the effects on the lipid content (LC) of oocytes and embryos and also verified the expression of 96 transcripts related to competence in matured COCs and 96 transcripts related to embryo quality in blastocysts. After IVM, LC was decreased in oocytes by NPPC while sildenafil did not affect LC in oocytes. The genes involved in lipid metabolism and lipid accumulation (DGAT1, PLIN2and PLIN3) were not affected in COCs after treatment during IVM, although the expression of PTX3 (a cumulus cells expansion biomarker) was increased and the hatched blastocyst rate was increased by NPPC during IVM. During IVM, sildenafil increased the mRNA relative abundance of HSF1 and PAF1 and decreased REST in blastocysts. The use of sildenafil in IVC increased the LC of blastocysts. The mRNA abundance in blastocysts produced during IVC with sildenafil was changed for ATF4, XBP1, DNMT3A, DNMT3B, COX2, and SOX2. Although NPPC reduced the LC of oocytes after IVM and upregulated markers for cumulus expansion, embryo production was not affected and the produced blastocysts were able to regain their LC after IVC. Finally, the use of sildenafil during IVC increased the cytoplasmic LC of embryos but did not affect embryo quality, as measured by analysis of 96 transcripts related to embryo quality.

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New technique to quantify the lipid composition of lipid droplets in porcine oocytes and pre-implantation embryos using Nile Red fluorescent probe

Cited by 57 publications

References 42 publications

Lipids and oocyte developmental competence: the role of fatty acids and β-oxidation

Lipids and oocyte developmental competence: the role of fatty acids and β-oxidation

The effect of maternal body condition on in vivo production of zygotes and behavior of delivered offspring in mice

Supplementing in vitro embryo production media by NPPC and sildenafil affect the cytoplasmic lipid content and gene expression of bovine cumulus-oocyte complexes and embryos

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