153 SUPPLEMENTATION WITH LINOLENIC ACID AND L-CARNITINE DURING IVM REDUCED THE EXPRESSION OF GENES RELATED TO LIPOGENESIS BUT DID NOT ALTER THE LIPID CONTENT AND CRYOTOLERANCE OF IN VITRO-PRODUCED EMBRYOS

2017 ◽  
Vol 29 (1) ◽  
pp. 185 ◽  
Author(s):  
B. C. S. Leao ◽  
N. A. S. Rocha Frigoni ◽  
P. C. Dall'Acqua ◽  
M. Ambrogi ◽  
G. B. Nunes ◽  
...  
Keyword(s):  
Gene Expression ◽  
Linolenic Acid ◽  
Lipid Content ◽  
Control Group ◽  
Intracellular Lipid ◽  
Chi Square ◽  
Sudan Black B ◽  
Expression Of Genes ◽  
The Impact

This study was conducted to evaluate the impact of supplementation during in vitro maturation (IVM) with linolenic acid (ALA), l-carnitine (L-car), or the combination of both supplements on the embryo intracellular lipid content and cryotolerance, as well as in the embryo expression of genes involved in lipid metabolism (lipogenesis regulation: SCD1, FASN, and SREBP1; and β-oxidation pathway: CPT1B and CPT2). Cumulus-oocyte complexes (n = 1076) were IVM for 22 h at 38.5°C and 5% CO2 in air, in TCM-199 medium with bicarbonate, hormones, and 10% FCS (control group), supplemented with 100 μM ALA (ALA group), 5 mM L-car (L-car group), or a combination of 100 μM ALA + 5 mM L-car (ALA + L-car group). After IVF, presumptive zygotes were in vitro cultured in SOFaa medium supplemented with 5 mg mL−1 BSA and 2.5% FCS, at 38.5°C and 5% CO2 in air during 7 days. Cleavage and blastocyst rates were evaluated on Day 3 and 7, respectively (IVF = Day 0). At Day 7, the blastocysts were stained with the lipophilic dye Sudan Black B (n = 60), vitrified/warmed (n = 260; Ingámed® protocol, Maringa-PR, Brazil), or collected for analysis of gene expression (n = 180). Embryonic development were analysed by ANOVA and the multiple comparisons of means were determined by Tukey’s test. The embryonic re-expansion data were subjected to chi-square test and the differences in gene expression among groups were evaluated by Duncan’s multiple range test (P < 0.05). Data are presented as means ± standard error means. There was no effect (P > 0.05) of the supplements used during IVM on cleavage (79.54 ± 2.76% to 82.16 ± 1.13%) and blastocyst rates (29.03 ± 3.07% to 30.46 ± 2.01%). Similarly, the intracellular lipid content in Day-7 blastocysts (1.03 ± 0.04 to 1.15 ± 0.07 pixels) and the embryonic cryotolerance, assessed by the re-expansion rates after 24 h (67.3 to 78.3%) hatching rates after 48 h (11.5 to 25.5%) of post-warming culture, were unaffected (P > 0.05) by the supplements of IVM medium. Although the treatments did not alter (P > 0.05) the expression of CPT1B and CPT2 genes, the expression of FASN gene was decreased (P < 0.05) in the ALA group and the expression of SREBP1 gene was decreased (P < 0.05) in the ALA and L-car groups. The expression of the gene SCD1 was reduced (P < 0.05) in all treatments compared with the control group. Thus, despite the lack of effects of the treatments performed during IVM on the intracellular lipid content and cryotolerance of the embryos derived from the treated oocytes, a reduction in the expression of genes related to lipogenesis was observed in Day-7 blastocysts. These results suggest that treatments performed in the oocytes during IVM may have prolonged effects, affecting the subsequent expression of genes in embryos. Further studies are needed to determine the mechanisms related to the differentiation of the oocyte machinery during maturation. Financial support was provided by FAPESP (#2012/10084–4 and #2013/07382–6).

scholarly journals Effects of Vitrification on the Blastocyst Gene Expression Profile in a Porcine Model

2021 ◽  
Vol 22 (3) ◽  
pp. 1222
Author(s):  
Cristina Cuello ◽  
Cristina A. Martinez ◽  
Josep M. Cambra ◽  
Inmaculada Parrilla ◽  
Heriberto Rodriguez-Martinez ◽  
...  
Keyword(s):  
Gene Expression ◽  
Gene Expression Analysis ◽  
Survival Rates ◽  
Control Group ◽  
P Value ◽  
Transcriptome Profile ◽  
Embryo Viability ◽  
The Impact

This study was designed to investigate the impact of vitrification on the transcriptome profile of blastocysts using a porcine (Sus scrofa) model and a microarray approach. Blastocysts were collected from weaned sows (n = 13). A total of 60 blastocysts were vitrified (treatment group). After warming, vitrified embryos were cultured in vitro for 24 h. Non-vitrified blastocysts (n = 40) were used as controls. After the in vitro culture period, the embryo viability was morphologically assessed. A total of 30 viable embryos per group (three pools of 10 from 4 different donors each) were subjected to gene expression analysis. A fold change cut-off of ±1.5 and a restrictive threshold at p-value < 0.05 were used to distinguish differentially expressed genes (DEGs). The survival rates of vitrified/warmed blastocysts were similar to those of the control (nearly 100%, n.s.). A total of 205 (112 upregulated and 93 downregulated) were identified in the vitrified blastocysts compared to the control group. The vitrification/warming impact was moderate, and it was mainly related to the pathways of cell cycle, cellular senescence, gap junction, and signaling for TFGβ, p53, Fox, and MAPK. In conclusion, vitrification modified the transcriptome of in vivo-derived porcine blastocysts, resulting in minor gene expression changes.

scholarly journals Circulating miRNA 887 is differentially expressed in ARDS and modulates endothelial function

2020 ◽  
Vol 318 (6) ◽  
pp. L1261-L1269 ◽  
Author(s):  
Andrew J. Goodwin ◽  
Pengfei Li ◽  
Perry V. Halushka ◽  
James A. Cook ◽  
Aman S. Sumal ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Function ◽  
In Silico Analysis ◽  
Leukocyte Migration ◽  
Differentially Expressed ◽  
Expression Of Genes ◽  
Cell Gene Expression ◽  
And Function ◽  
The Impact

Circulating microRNAs (miRNAs) can be taken up by recipient cells and have been recently associated with the acute respiratory distress syndrome (ARDS). Their role in host predisposition to the syndrome is unknown. The objective of the study was to identify circulating miRNAs associated with the development of sepsis-related ARDS and examine their impact on endothelial cell gene expression and function. We determined miRNA levels in plasma collected from subjects during the first 24 h of admission to a tertiary intensive care unit for sepsis. A miRNA that was differentially expressed between subjects who did and did not develop ARDS was identified and was transfected into human pulmonary microvascular endothelial cells (HPMECs). RNA sequencing, in silico analysis, cytokine expression, and leukocyte migration assays were used to determine the impact of this miRNA on gene expression and cell function. In two cohorts, circulating miR-887-3p levels were elevated in septic patients who developed ARDS compared with those who did not. Transfection of miR-887-3p into HPMECs altered gene expression, including the upregulation of several genes previously associated with ARDS (e.g., CXCL10, CCL5, CX3CL1, VCAM1, CASP1, IL1B, IFNB, and TLR2), and activation of cellular pathways relevant to the response to infection. Functionally, miR-887-3p increased the endothelial release of chemokines and facilitated trans-endothelial leukocyte migration. Circulating miR-887-3p is associated with ARDS in critically ill patients with sepsis. In vitro, miR-887-3p regulates the expression of genes relevant to ARDS and neutrophil tracking. This miRNA may contribute to ARDS pathogenesis and could represent a novel therapeutic target.

scholarly journals The Impact of Diet on Expression of Genes Involved in Innate Immunity in Goat Blood

2016 ◽  
Vol 8 (3) ◽  
pp. 1 ◽  
Author(s):  
Mulumebet Worku ◽  
Ahmed Abdalla ◽  
Sarah Adjei-Fremah ◽  
Hamid Ismail
Keyword(s):  
Gene Expression ◽  
Innate Immunity ◽  
Inflammatory Cytokines ◽  
Control Group ◽  
Colony Stimulating Factor ◽  
Sericea Lespedeza ◽  
Expression Of Genes ◽  
The Impact ◽  
Stimulating Factor ◽  
Pro Inflammatory Cytokines

<p>Sericea Lespedeza (SL), is a high-quality, low input forage that suppresses gastro-intestinal parasites in goats. The effect of dietary SL on the expression of genes involved in innate immunity in goats has not been established. The objective of this study was to evaluate the impact of a diet containing SL on the expression of genes involved in innate immunity in goat blood. Blood was collected by jugular venipuncture from goats fed a diet of 75% SL (n = 9) and a control group (n = 7), fed a SL free diet. Blood was used to evaluate expression of (CD-14, TLR-2, TLR-4, IL-10, IL-8, IL-2, INF-r, and TNF-a). Serum was extracted and used for evaluation of the secretion of pro-inflammatory cytokines (TNF-a, IFNr, granulocyte colony stimulating factor (GCSF), granulocyte-macrophage colony-stimulating factor (GMCSF), IL-1a, IL-8, IP-10 and RANTES) using a commercial ELISA kit. The level of gene expression of CD-14, TLR-2, TLR-4, IL-10, IL-8, IL-2, INF-r, and TNF-a was higher in treated animals compared to control. The <em>Sericea Lespedeza</em> diet affected the secretion of pro-inflammatory cytokines by increasing the serum levels of TNF-a, IFNr, GCSF, GMCSF, IL-1a, IP-10 (<em>P</em> &lt; 0.0002), and by decreasing (<em>P</em> &lt; 0.0001) IL-8 and RANTES in blood from goats fed SL. This suggests that dietary tannins modulate gene expression and may affect the goat's innate immune response in blood. Further research is needed to understand and harness the effect of dietary condensed tannins to modulate innate immunity in goats.</p>

203 SUPPLEMENTATION WITH LINOLENIC ACID, L-CARNITINE, ALONE OR ASSOCIATED, DURING IVM RESULTED IN DECREASE OF ROS LEVELS AND APOPTOTIC INDEX OF BOVINE IN VITRO PRODUCED EMBRYOS

2016 ◽  
Vol 28 (2) ◽  
pp. 232
Author(s):  
B. C. S. Leão ◽  
N. A. S. R. Frigoni ◽  
P. C. Dall'Acqua ◽  
M. Ambrogi ◽  
G. Z. Mingoti
Keyword(s):  
Reactive Oxygen Species ◽  
Embryonic Development ◽  
Linolenic Acid ◽  
Lipid Content ◽  
Reactive Oxygen ◽  
Antioxidant Effect ◽  
Apoptotic Index ◽  
Control Group ◽  
Oxygen Species

Supplementation of in vitro maturation (IVM) medium with linolenic acid (ALA) has been used in order to reduce oocyte lipid content and have beneficial effects on maturation and acquisition of competence for embryonic development. Besides the effect of reducing cellular lipid content, l-carnitine (l-car) has an antioxidant effect by reducing the levels of reactive oxygen species (ROS) and protecting cells from apoptosis. However, the association of ALA and l-car has never been tested. This study was conducted to evaluate the effects of supplementation of IVM medium of bovine oocytes with ALA, l-car or the association of both (ALA+l-car) on embryonic development and blastocysts reactive oxygen species (ROS) levels and occurrence of apoptosis. Cumulus-oocyte complexes (n = 2241, in 11 replicates) were matured during 22 h at 38.5°C and 5% CO2 in air, in TCM-199 medium with bicarbonate, hormones and 10% FCS (control group), also supplemented with 100 μM ALA group; or 5 mM l-car (l-car group); or 100 μM ALA associated with 5 mM l-car (ALA+l-car group). After fertilisation (Day 0), zygotes were cultured 7 days in SOF that was supplemented with 0.5% BSA and 2.5% FCS, in 5% CO2 in air at 38.5°C. The cleavage and blastocysts rates were evaluated, respectively, at Days 3 and 7. Blastocysts were stained with 5 mM of H2DCFDA (Molecular Probes, Invitrogen, Carlsbad, CA, USA) and TUNEL (In Situ Cell Death Detection Kit, Roche Applied Science, Boston, MA, USA), to evaluate the ROS levels and the blastomers apoptotic index, respectively. The ROS (n = 115) and TUNEL (n = 102) stained blastocysts were evaluated under an epifluorescence microscope (excitation 495 nm/510–550 nm and emission 404 nm/590 nm), and the ROS levels (expressed as arbitrary fluorescence units) were measured by Q-Capture Pro image software (Q Imaging, Surrey, BC, Canada). The fluorescence intensity values were subtracted from mean values of background in the images. The variables were analysed by ANOVA followed by Tukey’s test (P < 0.05) and data are presented as mean ± s.e.m. There was no effect (P > 0.05) of the supplements during IVM on cleavage and blastocysts rates (%), respectively, for control (81.1 ± 1.8 and 29.0 ± 3.1), ALA (80.5 ± 2.1 and 29.7 ± 2.3), l-car (79.5 ± 2.8 and 29.2 ± 2.3), and ALA+l-car (82.2 ± 1.1 and 30.5 ± 2.0) groups. The oocytes supplementation resulted in a decrease (P < 0.05) in ROS levels for ALA (0.84 ± 0.04), l-car (0.85 ± 0.03) and ALA+l-car (0.82 ± 0.02) groups, compared to the Control (1.00 ± 0.05). Consequently, the percentage of apoptotic blastomeres decreased (P < 0.05) after ALA (6.9 ± 1.0%), l-car (7.5 ± 1.2%) and ALA+l-car (4.6 ± 0.7%) supplementations, unlike to the Control group (12.0 ± 1.2%). In conclusion, the supplementation with ALA, l-car or ALA+l-car during IVM did not affect the blastocyst development, but led to a reduction in ROS levels and in the apoptotic index of such blastocysts. These findings may be due to some antioxidant effect of these supplements in the oocytes and/or the produced embryos. Financial support was through FAPESP (#2012/10084–4 and #2013/07382–6).

scholarly journals Lipid content and cryotolerance of in vitro-produced bovine embryos treated with forskolin before vitrification

2017 ◽  
Vol 37 (4) ◽  
pp. 395-400 ◽  
Author(s):  
Melissa Meneghel ◽  
Priscila Chediek Dall’Acqua ◽  
Marcela Ambrogi ◽  
Beatriz C.S. Leão ◽  
Nathália A.S. Rocha-Frigoni ◽  
...  
Keyword(s):  
Lipid Content ◽  
Culture Medium ◽  
Control Group ◽  
Bovine Embryos ◽  
Treatment Groups ◽  
Sudan Black B ◽  
Content Development ◽  
Pre Treatment

ABSTRACT: The aim of the present study was to evaluate the intracytoplasmic lipid content, development and cryotolerance of in vitro-produced bovine embryos treated with different concentrations of forskolin before vitrification. Embryos were produced from abattoir-derived ovaries and allocated into four groups. In the treatment groups, forskolin was added to the in vitro culture medium on Day 6 and incubated for 24 hours in one of the following concentrations: 2.5μM (Forsk 2.5 group), 5.0μM (Forsk 5.0 group) or 10.0μM (Forsk 10.0 group). Embryos from the control group were cultured without forskolin. On Day 7 of culture, the expanded blastocysts were stained with the lipophilic dye Sudan Black B for determination of the intracytoplasmic lipid content or were cryopreserved via the Vitri-Ingá® procedure. Although there were no significant differences (P>0.05) in the blastocyst rates between the Control group (44.9%) and the other treatments, the embryo production was lower (P<0.05) in Forsk 10.0 group (38.8%) compared to Forsk 2.5 (50.5%) and Forsk 5.0 (54.7%) groups. The intracytoplasmic lipid content (expressed in arbitrary units of pixels) in blastocysts from the Control group (1.00±0.03) was similar (P>0.05) to that found in Forsk 2.5 (0.92±0.03) and Forsk 10.0 groups (1.06±0.03) groups; however the lipid accumulation in blastocysts from Forsk 5.0 group (0.82±0.04) was lower than in the Control group (P<0.05). Based on these results, Forsk 5.0 treatment was tested for cryotolerance and it was observed that the blastocoel re-expansion rate evaluated 24 hours after warming was greater (P<0.05) in Forsk 5.0 group (72.2%) compared to the Control group (46.2%). In conclusion, pre-treatment with forskolin at a concentration of 5.0 μM for 24 hours before vitrification is effective in reducing the intracytoplasmic lipid content and, consequently, improves cryotolerance of IVP bovine embryos.

scholarly journals Bovine oocyte developmental competence and gene expression following co-culturing with ampullary cells: An experimental study

2021 ◽  
Author(s):  
Mehdi Azari ◽  
Mojtaba Kafi ◽  
Anise Asaadi ◽  
Zohreh Pakniat ◽  
Beheshteh Abouhamzeh
Keyword(s):  
Gene Expression ◽  
In Vitro Maturation ◽  
Fertilization Rate ◽  
Developmental Competence ◽  
Control Group ◽  
Sufficient Information ◽  
Nuclear Maturation ◽  
Oocyte Developmental Competence ◽  
The Impact

Background: There is no sufficient information on the impact of bovine ampullary oviductal epithelial cells (BAOECs) on in vitro oocyte maturation competence and gene expression. Objective: This study aimed to examine the oocyte developmental competence following co-culturing with a monolayer of fresh and frozen-thawed ampullary cells. Materials and Methods: Bovine cumulus-oocyte complexes (COCs) were distributed into three groups: control group; where in COCs were cultured in cell-free media for 24 hr and FML and FTML groups in which the COCs were cultured in maturation media for 18 hr and then transferred into a media containing fresh and frozen-thawed BAOECs monolayer, respectively (BAOECs were extracted from the oviducts of slaughtered cattle and were then cultured freshly or frozen-thawed) for a further 6 hr. After 24 hr, the expanded COCs were evaluated for nuclear maturation, fertilization rate, and gene expression (GDF9, StAR, CASP3, and FSHr). Results: Nuclear maturation rate in the FTML group was significantly higher than the control group (p = 0.02). The fertilization rate of FTML group was significantly higher than the control and FML groups (p = 0.05 and p = 0.03, respectively). In terms of gene expression, GDF9 were upregulated in the presence of the BAOECs during the last 6 hr of the in vitro maturation (p < 0.001). Furthermore, the expression of the StAR gene in the FTML group was higher than the other groups (p = 0.02). Conclusion: Ampullary cells co-culturing (especially frozen-thawed cells) for in vitro maturation of bovine oocytes yields encourages the results and demonstrates the beneficial effect of co-culture on gene expression and developmental competence. Key words: Ampulla, Bovine, Fertilization, Gene expression, IVM.

scholarly journals 164 EFFECT OF ACCUMULATION OF LIPIDS DURING IN VITRO CULTURE ON BOVINE BLASTOCYST YIELD AND FETAL DEVELOPMENT

2005 ◽  
Vol 17 (2) ◽  
pp. 233 ◽  
Author(s):  
J.A. Rooke ◽  
M. Ewen ◽  
G.J. McCallum ◽  
R.G. Watt ◽  
T.G. McEvoy
Keyword(s):  
Lipid Content ◽  
Fetal Development ◽  
Control Group ◽  
Chi Square ◽  
Crown Rump Length ◽  
Yield And Quality ◽  
Standard Procedures ◽  
Total Fatty Acids ◽  
Recorded Data

Previous research has shown that increased embryo lipid content, through culture in the presence of serum, is associated with reduced blastocyst yield and quality; provision of antioxidants ameliorates the effects of lipid accumulation (McEvoy et al. Reprod. Fertil. Dev. 16, 200). The present study extended these observations to assessment of fetal development from blastocysts which had accumulated lipid in vitro. Bovine oocytes, aspirated from abattoir-derived ovaries, were matured and fertilized using standard procedures. Cleaved zygotes were assigned to culture (5% CO2; 5% O2; 90% N2; 38.5°C) in four treatments: (1) synthetic oviductal fluid containing 0.3% bovine serum albumin and amino acids (SOF); (2) SOF plus supplementary bovine lipoproteins (2%, SOFLP); (3) SOF plus the antioxidant Trolox® (100 μM; 6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid, Sigma-Aldrich Co., Inc., Irvine, UK; SOFT); and (4) SOF plus lipoproteins and Trolox (SOFLPT). Blastocysts were synchronously transferred to recipient cattle (day 7; 23/treatment) together with a control group (n = 21) of artificially inseminated (AI) cattle. Reproductive tracts were recovered on Day 70 and fetal total and organ weights recorded. Data were analyzed by ANOVA (with fetal sex as covariate) and chi-square analyses. Culture in the presence of lipoproteins increased blastocyst total fatty acids (mean ± SD) from 98 ± 9.7 to 124 ± 7.3 ng/blastocyst. Blastocyst yields (%; 23 batches of ovaries) were reduced (P = 0.002) by inclusion of lipoproteins in culture (SOF, 22.0 ± 8.20; SOFLP, 16.4 ± 8.57; SOFT, 22.8 ± 9.03; SOFLPT, 24.2 ± 7.30) unless Trolox was present. Blastocyst grade was poorer (P < 0.001) after culture in the presence of lipoproteins irrespective of the presence of Trolox (SOF, 2.4 ± 0.43; SOFLP, 2.6 ± 0.45; SOFT, 2.4 ± 0.47; SOFLPT, 2.6 ± 0.40). Pregnancy rates (Day 70) were greater (P < 0.001) for AI (91%) than culture (26%) but were not affected by culture treatment. Although there were no differences in fetal weights between AI and IVC fetuses, SOFLP fetuses (Table 1) were lighter, had smaller relative liver weights, and had greater crown-rump length-to-weight ratios than other IVC fetuses. Therefore, IVC in conditions that increased blastocyst lipid content without adequate antioxidant protection reduced blastocyst yield and influenced fetal development. Table 1. Fetal weights (g) and relative liver weights (g/kg) and crown-rump lengths (CRL, mm/g) at day 70 SAC receives financial support from SEERAD.

scholarly journals 46 Effect of the addition of α-tocopherol to in vitro maturation media of bovine oocytes

2020 ◽  
Vol 98 (Supplement_2) ◽  
pp. 2-3
Author(s):  
Theisy P Acosta Pérez
Keyword(s):  
In Vitro Maturation ◽  
Cumulus Cells ◽  
Control Group ◽  
Chi Square ◽  
Cumulus Expansion ◽  
Minimum Concentration ◽  
Maturation Rate ◽  
Chi Square Test ◽  
System Data

Abstract α-tocopherol is known to be a powerful antioxidant, in this regard, it was added to bovine oocyte in vitro maturation media to evaluate its effect on oocyte maturation. Oocytes (n = 624) aspirated from ovaries of slaughtered cows were classified by quality and divided in four categories according to cytoplasm appearance and cumulus cells layers. Oocytes were washed in TCM-199 supplemented with fetal bovine serum (FBS) and FSH, then distributed in maturation media (TCM-199 supplemented with FBS, FSH and gentamicin). Three experimental groups of α-tocopherol (50, 100 and 200 mM) and a control group without α-tocopherol were used. Maturation was carried 22 h at 38.5°C in a 5% CO2 atmosphere. Oocytes were examined to determine cumulus expansion as categorical data (expansion or no expansion), as well as cumulus expansion Index (CEI). For CEI determination oocytes were graded 0 to 4, being 0 those with null expansion and 4 those with a noticeable cell expansion, then the number of oocytes were multiplied by the grade given and a sum of the totals was obtained, the new total was divided by the total of oocytes in the group and the result obtained corresponded to the CEI of the group. Results were analyzed with Chi Square test (for maturation rates) and an ANOVA (for the CEI) using the SAS system, data are presented as mean ± standard error. There was no statistical difference between control and α-tocopherol groups (P &gt;0.05). Numerically, the control group showed a higher maturation rate (100%) and obtained a higher CEI (2.44±0.20), followed by the 50 mM group (98.16%; 2.39±0.13), the groups 200 mM (97.40%; 2.00±0.14) and 100 mM (96.25%; 2.06±0.24) were the lowest. The addition of the minimum concentration (50 mM) of α-tocopherol to the maturation media could improve maturation rates without exposing oocytes to toxic effects.

scholarly journals Impact of Environmental Stressors on Gene Expression in the Embryo of the Italian Wall Lizard

2021 ◽  
Vol 11 (11) ◽  
pp. 4723
Author(s):  
Rosaria Scudiero ◽  
Chiara Maria Motta ◽  
Palma Simoniello
Keyword(s):  
Gene Expression ◽  
Protein Interactions ◽  
Membrane Trafficking ◽  
Translational Regulation ◽  
Cellular Protection ◽  
Terrestrial Vertebrate ◽  
Expression Of Genes ◽  
Stress Changes ◽  
Probable Consequence ◽  
The Impact

The cleidoic eggs of oviparous reptiles are protected from the external environment by membranes and a parchment shell permeable to water and dissolved molecules. As a consequence, not only physical but also chemical insults can reach the developing embryos, interfering with gene expression. This review provides information on the impact of the exposure to cadmium contamination or thermal stress on gene expression during the development of Italian wall lizards of the genus Podarcis. The results obtained by transcriptomic analysis, although not exhaustive, allowed to identify some stress-reactive genes and, consequently, the molecular pathways in which these genes are involved. Cadmium-responsive genes encode proteins involved in cellular protection, metabolism and proliferation, membrane trafficking, protein interactions, neuronal transmission and plasticity, immune response, and transcription regulatory factors. Cold stress changes the expression of genes involved in transcriptional/translational regulation and chromatin remodeling and inhibits the transcription of a histone methyltransferase with the probable consequence of modifying the epigenetic control of DNA. These findings provide transcriptome-level evidence of how terrestrial vertebrate embryos cope with stress, giving a key to use in population survival and environmental change studies. A better understanding of the genes contributing to stress tolerance in vertebrates would facilitate methodologies and applications aimed at improving resistance to unfavourable environments.

scholarly journals Cell–cell coupling and DNA methylation abnormal phenotypes in the after-hours mice

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Federico Tinarelli ◽  
Elena Ivanova ◽  
Ilaria Colombi ◽  
Erica Barini ◽  
Edoardo Balzani ◽  
...  
Keyword(s):  
Gene Expression ◽  
Dna Methylation ◽  
Neuronal Networks ◽  
Molecular Mechanisms ◽  
Ex Vivo ◽  
Neuronal Cultures ◽  
Functional Impairments ◽  
After Hours ◽  
The Impact

Abstract Background DNA methylation has emerged as an important epigenetic regulator of brain processes, including circadian rhythms. However, how DNA methylation intervenes between environmental signals, such as light entrainment, and the transcriptional and translational molecular mechanisms of the cellular clock is currently unknown. Here, we studied the after-hours mice, which have a point mutation in the Fbxl3 gene and a lengthened circadian period. Methods In this study, we used a combination of in vivo, ex vivo and in vitro approaches. We measured retinal responses in Afh animals and we have run reduced representation bisulphite sequencing (RRBS), pyrosequencing and gene expression analysis in a variety of brain tissues ex vivo. In vitro, we used primary neuronal cultures combined to micro electrode array (MEA) technology and gene expression. Results We observed functional impairments in mutant neuronal networks, and a reduction in the retinal responses to light-dependent stimuli. We detected abnormalities in the expression of photoreceptive melanopsin (OPN4). Furthermore, we identified alterations in the DNA methylation pathways throughout the retinohypothalamic tract terminals and links between the transcription factor Rev-Erbα and Fbxl3. Conclusions The results of this study, primarily represent a contribution towards an understanding of electrophysiological and molecular phenotypic responses to external stimuli in the Afh model. Moreover, as DNA methylation has recently emerged as a new regulator of neuronal networks with important consequences for circadian behaviour, we discuss the impact of the Afh mutation on the epigenetic landscape of circadian biology.

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