2 SPECIFIC FATTY ACID FOLLOW-UP REVEALS RUMEN-PROTECTED FAT SUPPLEMENTATION EFFECTS ON BOVINE OOCYTE QUALITY AND EMBRYO DEVELOPMENT

2014 ◽  
Vol 26 (1) ◽  
pp. 115 ◽  
Author(s):  
A. F. González-Serrano ◽  
C. R. Ferreira ◽  
V. Pirro ◽  
J. Heinzmann ◽  
K.-G. Hadeler ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Palmitic Acid ◽  
Embryo Development ◽  
Follicular Fluid ◽  
Oocyte Quality ◽  
Lipid Profiles ◽  
Dependent Manner ◽  
Fat Supplementation

Information on how supplementation of high-yield dairy cows with rumen-protected fat affects fertility in cattle herds is scarce. Here, Holstein-Friesian heifers (n = 84) received a supplement consisting of either rumen-protected conjugated linoleic acid (CLA; cis-9,trans-11-CLA and trans-10,cis-12-CLA) or stearic acid 18 : 0 (SA) on top of an isocaloric grass silage diet. Two supplementation doses were used (100 and 200 g d–1). Blood and follicular fluid were collected at the start and end of the supplementation period for analysis of cholesterol, insulin-like growth factor (IGF), and nonesterified fatty acids (NEFA), and for fatty acid profiling. Although cholesterol, IGF, and NEFA levels did not differ among experimental groups, lipid profiles in blood and follicular fluid were affected in a dose-dependent manner by both supplements. After 45 days of supplementation, oocytes were collected by ovum pick-up (OPU). The mRNA relative abundance of target genes (IGF1r, GJA1, FASN, SREBP1, and SCAP) was analysed in single in vitro- (24 h IVM) and in vivo-matured (collected by OPU 20 h after GnRH injection) oocytes and in vitro-produced blastocysts (Day 8) by qPCR (n = 6/group). Lipid profiling of individual oocytes from the CLA-supplemented (n = 37) and the SA-supplemented (n = 50) was performed by desorption electrospray ionization mass spectrometry (DESI-MS). Oocytes from the CLA-supplemented (n = 413) and the SA-supplemented (n = 350) groups were used for assessing maturation and blastocysts development rates. In immature oocytes, CLA supplementation led to an increase of triacylglycerol 52 : 3 [TAG (52 : 3)] and TAG (52 : 2), squalene, palmitic acid 16 : 0, and oleic acid 18 : 1, and decreased abundance of TAG (56 : 3), TAG (50 : 2) and TAG (48 : 1). In vitro-matured oocytes showed different lipid profiles, with increased abundances of TAG (52 : 3), and TAG (52 : 2) as well as phosphatidylinositol 34 : 1 [Plo (34 : 1)], whereas phosphatidylglycerol (34 : 1) [PG (34 : 1)] and palmitic acid 16 : 0 were less abundant in in vitro-matured oocytes. SCAP was significantly down-regulated in in vitro-matured oocytes from supplemented heifers compared with their in vivo-matured counterparts. Maturation (CLA = 74% v. SA = 67%) and blastocyst rates (CLA = 22.4% v. SA = 12.7%) were different among experimental groups. One-way ANOVA and the Tukey-Kramer test were applied for a multiple comparison of means (P-value ≤ 0.05 was considered as statistically significant). In conclusion, we demonstrate here that fatty acid monitoring along different compartments (i.e. blood system, follicular fluid, and intra-oocyte) after rumen-protected fat supplementation of dairy heifer diet reveals nutritional footprints on oocyte quality and embryo development. These results demonstrate the close relationship between nutrition and cattle herd's fertility and, at the same time, support the role of the bovine model for understanding nutritional-dependent fertility impairments.

180 FATTY ACID COMPOSITION IN FOLLICULAR FLUID OF PREPUBERTAL GOAT OVARIES IN WINTER AND AUTUMN

2015 ◽  
Vol 27 (1) ◽  
pp. 181
Author(s):  
M. T. Paramio ◽  
M. Roura ◽  
S. Hammami ◽  
D. Izquierdo ◽  
M. G. Catalá
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Embryo Development ◽  
Follicular Fluid ◽  
Oocyte Quality ◽  
Blastocyst Stage ◽  
Omega 3 ◽  
Chromatography Analysis ◽  
Sperm Penetration

Fatty acids (FA) in follicular fluid (FF) play an important role on oocyte quality and embryo development (Fouladi-Nashta et al. 2007 Biol. Reprod. 77, 9–17). In our laboratory, we have shown in prepubertal goat differences in the percentage of blastocysts produced in vitro according to season. Thus, we have found in winter 15.8% and in autumn a decrease up to 4.7% of blastocysts that were produced from oocytes of 1 month old suckling Murciano-Granadina goat females and IVF with fresh semen. The aim of this study was to analyse composition of FF in order to find an explanation to seasonal changes in in vitro embryo production. Ovaries were recovered in winter and autumn from 1 month suckling goats (Murciano-Granadina) from a local slaughterhouse and the FF of all visible follicles was recovered using a sterile syringe. Each sample containing a pool of FF of different ovaries was frozen at –80°C until chromatography analysis. For the FA analysis, the Sukhija and Palmquist (1988 J. Agric. Food Chem. 36, 1202–1206) protocol with some adaptations was used. Briefly, 200 μL of FF sample was vortexed for 60 s with 250 μL of toluene and 1 mL of HCL (5%) and then warmed in a water bath for 1 h at 70°C. Subsequently 1.25 mL of K2CO3 (12%) and 500 μL of toluene was added, vortexed for 30 s and centrifuged for 5 min (3000 rpm). Finally the supernatant was recovered and dried with Na2SO4. The extracted samples were maintained in –20°C until gas chromatographic analysis (123–2362, Agilent Technologies Inc., Santa Clara, CA). The results in Table 1 express the mean of 3 replicates of follicular fluid pool as micromolar concentration of FA in FF. The FA profile in FF showed significant higher concentrations of α-linolenic (C18:3n3), eicosapentaenoic (EPA), docosahexaenoic (DHA), and omega-3 (n-3 polyunsaturated fatty acid; PUFA) in winter compared to autumn. This could be indicating that these PUFA have a positive effect on oocyte quality because of the higher embryo development of these oocytes during winter. Studies in our laboratory have shown that sperm penetration and normal zygotes were similar in both seasons even though the blastocyst yield was statistically higher in winter. We can speculate that fatty acids in the follicular environment are affecting the oocyte quality, increasing the possibility of reaching the blastocyst stage in prepubertal goat according to season. Further studies should be done to reach a more accurate conclusion. Table 1.Concentration (µM) of fatty acids in FF of prepubertal goat during winter and autumn (3 replicates)

scholarly journals Amino acid and fatty acid composition of follicular fluid as predictors of in-vitro embryo development

2008 ◽  
Vol 16 (6) ◽  
pp. 859-868 ◽  
Author(s):  
KD Sinclair ◽  
LA Lunn ◽  
WY Kwong ◽  
K Wonnacott ◽  
RST Linforth ◽  
...  
Keyword(s):  
Fatty Acid Composition ◽  
Fatty Acid ◽  
Amino Acid ◽  
Embryo Development ◽  
Acid Composition ◽  
Follicular Fluid

scholarly journals Antimycobacterial action of thiolactomycin: an inhibitor of fatty acid and mycolic acid synthesis.

1996 ◽  
Vol 40 (12) ◽  
pp. 2813-2819 ◽  
Author(s):  
R A Slayden ◽  
R E Lee ◽  
J W Armour ◽  
A M Cooper ◽  
I M Orme ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Fatty Acid Synthase ◽  
Acyl Carrier Protein ◽  
Mycolic Acid ◽  
Carrier Protein ◽  
Type I ◽  
Dependent Manner ◽  
Fas Ii

Thiolactomycin (TLM) possesses in vivo antimycobacterial activity against the saprophytic strain Mycobacterium smegmatis mc2155 and the virulent strain M. tuberculosis Erdman, resulting in complete inhibition of growth on solid media at 75 and 25 micrograms/ml, respectively. Use of an in vitro murine macrophage model also demonstrated the killing of viable intracellular M. tuberculosis in a dose-dependent manner. Through the use of in vivo [1,2-14C]acetate labeling of M. smegmatis, TLM was shown to inhibit the synthesis of both fatty acids and mycolic acids. However, synthesis of the shorter-chain alpha'-mycolates of M. smegmatis was not inhibited by TLM, whereas synthesis of the characteristic longer-chain alpha-mycolates and epoxymycolates was almost completely inhibited at 75 micrograms/ml. The use of M. smegmatis cell extracts demonstrated that TLM specifically inhibited the mycobacterial acyl carrier protein-dependent type II fatty acid synthase (FAS-II) but not the multifunctional type I fatty acid synthase (FAS-I). In addition, selective inhibition of long-chain mycolate synthesis by TLM was demonstrated in a dose-response manner in purified, cell wall-containing extracts of M. smegmatis cells. The in vivo and in vitro data and knowledge of the mechanism of TLM resistance in Escherichia coli suggest that two distinct TLM targets exist in mycobacteria, the beta-ketoacyl-acyl carrier protein synthases involved in FAS-II and the elongation steps leading to the synthesis of the alpha-mycolates and oxygenated mycolates. The efficacy of TLM against M. smegmatis and M. tuberculosis provides the prospects of identifying fatty acid and mycolic acid biosynthetic genes and revealing a novel range of chemotherapeutic agents directed against M. tuberculosis.

Effect of butafosfan supplementation during oocyte maturation on bovine embryo development

Zygote ◽  
2019 ◽  
Vol 27 (05) ◽  
pp. 321-328
Author(s):  
Lucas Teixeira Hax ◽  
Joao Alveiro Alvarado Rincón ◽  
Augusto Schneider ◽  
Lígia Margareth Cantarelli Pegoraro ◽  
Letícia Franco Collares ◽  
...  
Keyword(s):  
Gene Expression ◽  
Embryo Development ◽  
Oocyte Maturation ◽  
Cumulus Cells ◽  
Oocyte Quality ◽  
Bovine Embryo ◽  
Cleavage Rate ◽  
Nuclear Maturation

SummaryAround 60–80% of oocytes maturated in vivo reached competence, while the proportion of maturation in vitro is rarely higher than 40%. In this sense, butafosfan has been used in vivo to improve metabolic condition of postpartum cows, and can represent an alternative to increase reproductive efficiency in cows. The aim of this study was to evaluate the addition of increasing doses of butafosfan during oocyte maturation in vitro on the initial embryo development in cattle. In total, 1400 cumulus–oocyte complexes (COCs) were distributed in four groups and maturated according to supplementation with increasing concentrations of butafosfan (0 mg/ml, 0.05 mg/ml, 0.1 mg/ml and 0.2 mg/ml). Then, 20 oocytes per group were collected to evaluate nuclear maturation and gene expression on cumulus cells and oocytes and the remaining oocytes were inseminated and cultured until day 7, when blastocysts were collected for gene expression analysis. A dose-dependent effect of butafosfan was observed, with decrease of cleavage rate and embryo development with higher doses. No difference between groups was observed in maturation rate and expression of genes related to oocyte quality. Our results suggest that butafosfan is prejudicial for oocytes, compromising cleavage and embryo development.

scholarly journals Berberine Suppressed Tumor Growth through Regulating Fatty Acid Metabolism and Triggering Cell Apoptosis via Targeting FABPs

2020 ◽  
Vol 2020 ◽  
pp. 1-16 ◽  
Author(s):  
Lingli Li ◽  
Ze Peng ◽  
Qian Hu ◽  
Lijun Xu ◽  
Xin Zou ◽  
...  
Keyword(s):  
Gastric Cancer ◽  
Fatty Acid ◽  
Protein Expression ◽  
Cell Viability ◽  
Human Gastric Cancer ◽  
Dependent Manner ◽  
Xenograft Models ◽  
Induced Apoptosis

Aim. To further investigate the mechanism behind the antitumor properties of berberine regarding lipid metabolism. Methods. Cell viability, proliferation, and apoptosis assays were performed to determine the antigrowth effects of berberine in vitro. Ectopic xenograft models in Balb/c nude mice were established to determine the antitumor effects of berberine in vivo. Results. Berberine inhibited cell viability and proliferation of MGC803 human gastric cancer cell lines in a time- and dose-dependent manner. Berberine induced apoptosis of MGC803 and increased the apoptotic rate with higher doses. Berberine induced the accumulation of fatty acid of MGC803 and suppressed the protein expression of FABPs and PPARα. The FABP inhibitor BMS309403 recapitulated the effects of berberine on MGC803 cells. In the xenograft model, berberine significantly decreased the tumor volume and tumor weight and induced apoptosis in tumor tissues. Berberine significantly elevated the fatty acid content and inhibited the expression of FABPs and PPARα in the MGC803 xenograft models. Conclusion. Berberine exerted anticancer effects on human gastric cancer both in vitro and in vivo by inducing apoptosis, which was due to the reduced protein expression of FABPs and the accumulation of fatty acid.

Synaptic Connectivity and Cortical Maturation Are Promoted by the ω-3 Fatty Acid Docosahexaenoic Acid

2019 ◽  
Vol 30 (1) ◽  
pp. 226-240
Author(s):  
Beatrice E Carbone ◽  
Malik Abouleish ◽  
Katherine E Watters ◽  
Seth Vogel ◽  
Adema Ribic ◽  
...  
Keyword(s):  
Visual Acuity ◽  
Fatty Acid ◽  
Docosahexaenoic Acid ◽  
Neurite Growth ◽  
Light Sensitivity ◽  
Oscillatory Behavior ◽  
Synaptic Connectivity ◽  
Dependent Manner

Abstract Brain development is likely impacted by micronutrients. This is supported by the effects of the ω-3 fatty acid docosahexaenoic acid (DHA) during early neuronal differentiation, when it increases neurite growth. Aiming to delineate DHA roles in postnatal stages, we selected the visual cortex due to its stereotypic maturation. Immunohistochemistry showed that young mice that received dietary DHA from birth exhibited more abundant presynaptic and postsynaptic specializations. DHA also increased density and size of synapses in a dose-dependent manner in cultured neurons. In addition, dendritic arbors of neurons treated with DHA were more complex. In agreement with improved connectivity, DHA enhanced physiological parameters of network maturation in vitro, including bursting strength and oscillatory behavior. Aiming to analyze functional maturation of the cortex, we performed in vivo electrophysiological recordings from awake mice to measure responses to patterned visual inputs. Dietary DHA robustly promoted the developmental increase in visual acuity, without altering light sensitivity. The visual acuity of DHA-supplemented animals continued to improve even after their cortex had matured and DHA abolished the acuity plateau. Our findings show that the ω-3 fatty acid DHA promotes synaptic connectivity and cortical processing. These results provide evidence that micronutrients can support the maturation of neuronal networks.

scholarly journals 170 FOLLICULAR FLUID CONCENTRATION AND OOCYTE QUALITY FROM TOGGENBURG GOATS FED WITH UREA

2009 ◽  
Vol 21 (1) ◽  
pp. 184
Author(s):  
E. A. M. Amorim ◽  
L. S. Amorim ◽  
C. A. A. Torres ◽  
J. D. Guimãres ◽  
J. F. Fonseca ◽  
...  
Keyword(s):  
Follicular Fluid ◽  
Follicular Development ◽  
Cumulus Cells ◽  
Oocyte Quality ◽  
Urea Concentration ◽  
Follicular Aspiration ◽  
Randomized Design ◽  
Fluid Concentration

Protein and urea concentrations impair oocyte and embryo development in vivo and in vitro through an unclear mechanism. A possible way to understand this process is to determine the concentration of hormones and metabolites in follicular fluid associated with normal development. The objective of this study was to determine the effect of dietary urea levels on follicular fluid concentration of hormones and metabolites and oocyte quality. A trial was conducted with 9 nonpregnant and nonlactating Saanen goats, which had been distributed in a randomized design and fed with diets with 0 (n = 4) and 2.4% of urea in the total dry matter (DM) of the diet (n = 5). Before follicle aspiration by laparotomy, the goats were synchronized by inserting intravaginal sponges containing 60 mg of acetate medroxyprogesterone (Progespon®, Sintex) for 10 days followed by 125 μg of cloprostenol (Ciosin® Coopers) 48 h before the removal of the sponge. The sponge was removed immediately before the follicular aspiration. The follicular development was stimulated with 70 mg of NIH-FSH-P1 (Folltropin V® Vetrepharm) i.m., and 300 IU of eCG i.m., (Novormon® Sintex) given 36 h before the follicular aspiration. Fluid from the 2 lartest follicles of each ovary were analyzed to determine the concentration of estradiol, progesterone, and testosterone by quimioluminesence, and glucose and urea concentrations were measured by enzymatic kit. The other follicles in each ovary were aspired with new needles and syringes and the oocyte quality was recorded. Oocytes were classified according to cytoplasma aspect and number of granulosa cells: Class A (dark cytoplasm and uniform aspect) with 3 (AMG) and <3 layers of cumulus cells (AmG); class B (cytoplasm with color alterations, desuniform aspect and vacuoles) with 3 (BMG) and <3 layers of cumulus cells (BmG); number of partially denuded oocytes (PD) and number of denuded oocytes (DO). Data were analyzed by ANOVA and treatment difference separated by SNK test. Follicular fluid estradiol concentration was lower in goats fed with urea (4.02 ± 0.16; 4.97 ± 0.18 ng mL–1; P < 0.05), progesterone concentration did not differ between treatments (2.48 ± 0.58; 3.37 ± 0.52 ng mL–1; P > 0.05), testosterone concentration was lower in the control animals (1.17 ± 0.48; 3.20 ± 0.43 ng mL–1; P < 0.05). The glucose (91.44 ± 3.60; 84.78 ± 5.58 mg dL–1) and urea concentration (23.04 ± 1.06; 18.00 ± 2.35) were greater in the animals treated with 2.4% compared with 0% of urea (P < 0.05), respectively. The number of oocytes in the different categories was not affected by treatment (P > 0.05): AMG 1.20 ± 1.09 v. 0.50 ± 0.57, AmG 4.20 ± 2.16 v. 3.50 ± 3.10, BMG 0.40 ± 0.54 v. 0.25 ± 0.50, BmG 1.40 ± 0.54 v. 1.75 ± 1.25, DO 10.20 ± 3.76 v. 11.50 ± 5.44, in the 0 and 2.4% of urea groups respectively. Only the number of PD (1.60 ± 0.54 v. 3.50 ± 1.91) recovered from animals treated with 2.4% was greater than in controls (P < 0.05). The hormone and metabolites concentration in follicular fluid as well as the oocyte quality was affected by the urea concentration of the diet. Supported by grant from: CNPq, FAPEMIG, Shering Plough®, Tecnopec®, Carbogel®.

Growth differentiation factor 8 regulates SMAD2/3 signaling and improves oocyte quality during porcine oocyte maturation in vitro†

2019 ◽  
Vol 101 (1) ◽  
pp. 63-75 ◽  
Author(s):  
Junchul David Yoon ◽  
Seon-Ung Hwang ◽  
Mirae Kim ◽  
Yubyeol Jeon ◽  
Sang-Hwan Hyun
Keyword(s):  
Follicular Fluid ◽  
Transforming Growth Factor ◽  
Oocyte Quality ◽  
Mitogen Activated Protein Kinase ◽  
Specific Gene ◽  
Paracrine Factor ◽  
Differentiation Factor ◽  
Porcine Oocyte

Abstract Growth differentiation factor 8 (GDF8), also known as myostatin, is a member of the transforming growth factor-β (TGF-β) family and has been identified as a strong physiological regulator of muscle differentiation. Recently, the functional role of GDF8 in reproductive organs has received increased interest following its detection in the human placenta and uterus. To investigate the effects of GDF8 during porcine oocyte in vitro maturation (IVM), we assessed the quality of matured oocytes. Furthermore, we investigated the specific gene transcription and protein activation levels in oocytes and cumulus cells after IVM and subsequent embryonic development after in vitro fertilization and parthenogenetic activation. Prior to these experiments, the concentration of GDF8 in porcine follicular fluid was determined. During the entire IVM period, 1.3 ng/mL GDF8 and its signaling inhibitor SB431542 (SB) at 5 μM were added as control, SB, SB + GDF8, and GDF8 groups, respectively. Our results demonstrate that supplementation with GDF8 during porcine oocyte IVM enhanced both meiotic and cytoplasmic maturation, with altered transcriptional patterns, via activation of Sma- and Mad-related protein 2/3 (SMAD2/3). Using the pharmacological inhibitor SB431542, we demonstrated that inhibition of GDF8-induced Smad2/3 signaling reduces matured oocyte quality. In conclusion, for the first time, we demonstrated paracrine factor GDF8 in porcine follicular fluid in vivo. Furthermore, we showed that GDF8 supplementation improved mature oocyte quality by regulating p38 mitogen-activated protein kinase phosphorylation and intracellular glutathione and reactive oxygen species levels during porcine IVM.

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