The Effect of 17α-Ethynilestradiol and GPER1 Activation on Body and Muscle Growth, Muscle Composition and Growth-Related Gene Expression of Gilthead Seabream, Sparus aurata L
Abstract
:1. Introduction
2. Results
2.1. Body Parameters, Cumulative Intake and Survival
2.1.1. Body Parameters
2.1.2. Cumulative Intake and Survival
2.1.3. Fish Gonad Structure
2.2. Muscle Parameters
2.2.1. Muscle Parameters According to the Gonad Structure
2.2.2. Histochemical Characterization of the Muscle Fibers
2.3. Muscle Composition and Textural Hardness
Muscle Composition and Textural Hardness According to Gonad Structure
2.4. Growth-Related Gene Expression
3. Discussion
3.1. Body Growth and Survival
3.2. Muscle Growth Parameters, Muscle Enzymatic Activity and Textural Hardness
3.3. Muscle Protein and Fat
3.4. Growth-Related Gene Expression
4. Material and Methods
4.1. Animals and Management
4.2. Experimental Treatments and Quantification of Cumulative Intake and Final Survival
4.3. Sampling and Measurement of Body Parameters
4.4. Quantitative Analysis of Muscle Growth and Histochemical Techniques
4.5. Analysis of the Composition and Texture of the Fillet
4.6. Gene Expression Analysis
4.7. Statistical Analysis
5. Conclusions
- GPER1 activation and EE2 treatment reduced body growth in the short and long term, respectively.
- EE2 treatment, and to a lesser extent G1 treatment, reduced muscle growth (transverse area of the muscle) in the long term and was correlated with a high expression of key growth muscle-related genes in the EE2 group, such as mfbx (upregulated gene in catabolic signals) and mstn2 (negative regulator gene of the muscle growth).
- EE2 treatment and GPER1 activation increased the hypertrophy and glycolytic activity of the white muscle fibers, whereas fibrillary hyperplasia and muscle fiber density showed the contrary tendency, indicating immediate and persistent long-lasting effects on muscle cellularity.
- EE2 treatment and GPER1 activation reduced the textural firmness, showing a negative correlation with the size of the muscle fibers.
- EE2 exposure reduced muscle fat and was related with the predominance of ovarian area in the gonads of EE2-treated fish.
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Stage | Groups | BW (g) | GW (g) | BL (cm) | CF (100 × g cm−3) | HSI (%) | GSI (%) |
---|---|---|---|---|---|---|---|
First (on day 45 of treatment) | C | 314.4 a ± 8.5 | 286.86 a ± 7.41 | 26.6 a ± 0.2 | 1.68 ab ± 0.1 | 2.01 a ± 0.08 | 0.29 a ± 0.06 |
EE2 | 292.7 a ± 8.76 | 265.8 a ± 7.8 | 26.75 a ± 0.2 | 1.53 b ± 0.01 | 2.53 a ± 0.22 | 0.26 a ± 0.1 | |
G1 | 288.6 a ± 6.41 | 262.8 a ± 5.98 | 25.67 b ± 0.3 | 1.71 a ± 0.05 | 2.4 a ± 0.06 | 0.19 a ± 0.04 | |
Second (79 days after treatment cessation) | C | 374.9 a ± 9.85 | 344.2 a ± 9.54 | 28.42 a ± 0.47 | 1.6 a ± 0.06 | 1.5 a ± 0.3 | 0.22 a ± 0.04 |
EE2 | 300.23 b ± 12.16 | 292.6 b ± 7.12 | 27.17 b ± 0.21 | 1.5 a ± 0.08 | 1.87 a ± 0.57 | 0.2 a ± 0.03 | |
G1 | 366.03 a ± 9.09 | 336.2 a ± 9.18 | 27.92 ab ± 0.2 | 1.7 a ± 0.03 | 1.28 a ± 0.06 | 0.23 a ± 0.03 | |
Third (122 days after treatment cessation) | C | 428.23 ab ± 8.85 | 397.5 ab ± 8.24 | 29.0 a ± 0.36 | 1.76 a ± 0.05 | 0.93 a ± 0.05 | 0.22 a ± 0.05 |
EE2 | 384.38 b ± 13.66 | 355.47 b ± 12.99 | 28.5 a ± 0.22 | 1.66 a ± 0.04 | 0.91 a ± 0.004 | 0.22 a ± 0.03 | |
G1 | 471.57 a ± 23.7 | 436.9 a ± 21.91 | 29.6 a ± 0.42 | 1.82 a ± 0.05 | 0.95 a ± 0.05 | 0.18 a ± 0.02 |
Stage | Groups | W (mm2) | R (mm2) | A (μm2) | D (μm) | N | Dens |
---|---|---|---|---|---|---|---|
First (on day 45 of treatment) | C | 1251.2 a ± 35.6 | 204.5 a ± 9.6 | 5246.52 a ± 248.5 | 74.5 a ± 2.28 | 241,808.4 a ± 15,472.2 | 192.64 a ± 8.6 |
EE2 | 1327.1 a ± 49.2 | 224.75 a ± 9.4 | 5676.8 a ± 137.5 | 77.2 a ± 0.85 | 233,501.89 a ± 4373.5 | 176.69 a ± 4.46 | |
G1 | 1281.9 a ± 25.5 | 223.96 a ± 8.4 | 5773.6 a ± 265.4 | 78.8 a ± 1.73 | 223,892.97 a ± 9215.5 | 174.9 a ± 7.43 | |
Second (79 days after treatment cessation) | C | 1632.32 a ± 38.46 | 307.69 a ± 20.04 | 6825.29 a ± 440.66 | 85.40 a ± 3.2 | 244,515.14 a ± 17,359.1 | 149.7 a ± 9.8 |
EE2 | 1580.99 a ± 50.36 | 287.15 a ± 11.44 | 6871.26 a ± 492.99 | 85.43 a ± 2.9 | 236,680.22 a ± 19,501.24 | 149.03 a ± 9.9 | |
G1 | 1608.37 a ± 67.2 | 318.86 a ± 10.75 | 7266.59 a ± 314.0 | 88.3 a ± 1.95 | 223,948.69 a ± 14,958.87 | 138.87 a ± 5.85 | |
Third (122 days after treatment cessation) | C | 2166.78 a ± 62.8 | 377.6 a ± 18.5 | 6853.4 a ± 526.3 | 82.4 a ± 3.4 | 323,080.1 a ± 20,136.3 | 149.8 a ± 10.2 |
EE2 | 1924.18 a ± 80.9 | 340.3 a ± 10.7 | 8813.9 a ± 994.3 | 96.0 a ± 5.7 | 235,558.5 a ± 34,306.7 | 120.4 a ± 12.78 | |
G1 | 2063.9 a ± 90.1 | 374.6 a ± 12.9 | 7916.96 a ± 216.8 | 89.5 a ± 1.19 | 262,117.11 a ± 15,718.8 | 126.8 a ± 3.4 |
Stage | Groups | Protein (%) | Fat (%) | Hardness (N) |
---|---|---|---|---|
First (on day 45 of treatment) | C | 22.77 a ± 0.29 | 1.63 ab ± 0.1 | 29.42 a ± 1.7 |
EE2 | 22.50 a ± 0.33 | 1.28 b ± 0.04 | 29.21 a ± 1.14 | |
G1 | 22.30 a ± 0.42 | 1.67 a ± 0.12 | 23.87 a ± 2.43 | |
Second (79 days after treatment cessation) | C | 22.45 a ± 0.14 | 3.52 a ± 0.59 | 49.29 a ± 2.78 |
EE2 | 21.87 a ± 0.2 | 2.93 a ± 0.36 | 46.7 a ± 2.55 | |
G1 | 22.38 a ± 0.34 | 3.47 a ± 0.33 | 47.14 a ± 4.87 | |
Third (122 days after treatment cessation) | C | 21.3 a ± 0.51 | 9.43 a ± 1.73 | 58.06 a ± 8.61 |
EE2 | 22.6 a ± 0.17 | 5.38 a ± 0.37 | 53.8 a ± 6.64 | |
G1 | 22.03 a ± 0.31 | 7.18 a ± 1.1 | 56.16 a ± 6.93 |
Genes | BW | BL | W | R | T | Fat | Prot | H | A | D | N | Dens |
---|---|---|---|---|---|---|---|---|---|---|---|---|
mstn2 | 0.1 | 0.1 | 0.3 · | 0.4 | 0.3 * | 0.3 · | −0.3 | 0.4 ** | 0.5 ** | 0.4 ** | −0.2 | −0.5 ** |
capn1 | −0.3 · | −0.3 · | −0.4 ** | −0.5 ** | −0.5 ** | −0.1 | −0.1 | −0.2 | −0.3 * | −0.3 * | 0 | 0.4 * |
mafbx | −0.5 ** | −0.3 · | −0.4 * | −0.5 ** | −0.4 * | −0.2 | −0.2 | −0.4 * | −0.4 * | −0.4 ** | −0.4 * | 0.4 * |
myod | −0.2 | −0.2 | −0.3 · | −0.2 | −0.3 · | −0.2 | 0.1 | −0.2 | −0.1 | −0.1 | −0.2 | 0.1 |
Gene | ID | Primers (5′–3′) | Tm (°C) | E (%) | Size (bp) |
---|---|---|---|---|---|
myod2 | ENSSAUG00010004893 | F:CACTACAGCGGGGATTCAGAC | 83.3 | 99 | 149 |
R:CGTTTGCTTCTCCTGGACTC | |||||
mstn2 | ENSSAUG00010004791 | F:ACCTGGTGAACAAAGCCAAC | 84.8 | 99 | 201 |
R:TGCGGTTGAAGTAGAGCATG | |||||
capn1 | ENSSAUG00010025995 | F:CCTACGAGATGAGGATGGCT | 87 | 95 | 114 |
R:AGTTGTCAAAGTCGGCGGT | |||||
mafbx | ENSG00000156804 | F:GGTGCAACTTTCTGGGTTGT | 85 | 97 | 105 |
R:GGTCACCTGGAGTGGAAGAA |
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Ayala, M.D.; Gómez, V.; Cabas, I.; García Hernández, M.P.; Chaves-Pozo, E.; Arizcun, M.; Garcia de la Serrana, D.; Gil, F.; García-Ayala, A. The Effect of 17α-Ethynilestradiol and GPER1 Activation on Body and Muscle Growth, Muscle Composition and Growth-Related Gene Expression of Gilthead Seabream, Sparus aurata L. Int. J. Mol. Sci. 2021, 22, 13118. https://doi.org/10.3390/ijms222313118
Ayala MD, Gómez V, Cabas I, García Hernández MP, Chaves-Pozo E, Arizcun M, Garcia de la Serrana D, Gil F, García-Ayala A. The Effect of 17α-Ethynilestradiol and GPER1 Activation on Body and Muscle Growth, Muscle Composition and Growth-Related Gene Expression of Gilthead Seabream, Sparus aurata L. International Journal of Molecular Sciences. 2021; 22(23):13118. https://doi.org/10.3390/ijms222313118
Chicago/Turabian StyleAyala, Maria D., Victoria Gómez, Isabel Cabas, María P. García Hernández, Elena Chaves-Pozo, Marta Arizcun, Daniel Garcia de la Serrana, Francisco Gil, and Alfonsa García-Ayala. 2021. "The Effect of 17α-Ethynilestradiol and GPER1 Activation on Body and Muscle Growth, Muscle Composition and Growth-Related Gene Expression of Gilthead Seabream, Sparus aurata L" International Journal of Molecular Sciences 22, no. 23: 13118. https://doi.org/10.3390/ijms222313118