The In Vitro Evaluation of Rooster Semen Pellets Frozen with Dimethylacetamide

Overview

Journal Animals (Basel)

Date 2023 May 27

PMID 37238034

Authors

Shaimaa K Hamad

Ahmed M Elomda

Yanyan Sun

Yunlei Li

Yunhe Zong

Jilan Chen

Ahmed O Abbas

Farid K R Stino

Ali Nazmi

Gamal M K Mehaisen

Affiliations

Soon will be listed here.

Abstract

Sperm cryopreservation is an effective technique for conserving animal genetic diversity and transmitting superior genetic backgrounds, maintained via a non-invasive sampling and collection of huge quantities of sperm. Nevertheless, cryopreservation in avian species is not commercially viable because of the rooster sperm's susceptibility to damage. This study aims to estimate the impact of dimethylacetamide (DMA) as a cryoprotectant at different levels (3%, 6%, or 9%) on the post-thawed sperm quality, motility, antioxidant-biomarkers, and the expression of anti-freeze related genes. Semen samples were collected twice a week from twelve roosters aged 40 wk, weighing 3400 ± 70 g, and belonging to the Cairo-B2 chicken strain. Fresh semen samples were rapidly appraised, pooled, diluted with two volumes of a basic extender, and divided equally into three groups. The diluted groups were chilled at -20 °C for 7 min, then gently supplemented with 3, 6, or 9% pre-cooled DMA and equilibrated at 5 °C for a further 10 min. Semen pellets were formed by pipetting drops 7 cm above liquid nitrogen (LN), which were then kept inside cryovials in the LN. Thawing was performed 2 months later by taking 3-4 pellets of the frozen semen into a glass tube and warming it in a water bath for 8 s at 60 °C. The results showed that 3% DMA increased the proportion of total motile sperm, progressivity, viability, and plasma membrane integrity (%) compared to the 6% and 9% DMA groups. The lipid peroxidation and antioxidant enzyme activity were improved in the 3% group. At the same time, some anti-freeze-related genes' (including ras homolog family member A (RHOA), heat shock protein 70 (HSP70), and small nuclear ribonucleoprotein polypeptide A (SNRPA1)) expressions were upregulated within the 3% DMA group relative to other groups. In conclusion, the 3% DMA group maintained higher post-thawed sperm quality than the other tested groups.

References

Abouelezz F, Castano C, Toledano-Diaz A, Esteso M, Lopez-Sebastian A, Campo J . Effect of the interaction between cryoprotectant concentration and cryopreservation method on frozen/thawed chicken sperm variables. Reprod Domest Anim. 2014; 50(1):135-41. DOI: 10.1111/rda.12464. View

Murugesan S, Mahapatra R . Cryopreservation of Ghagus chicken semen: Effect of cryoprotectants, diluents and thawing temperature. Reprod Domest Anim. 2020; 55(8):951-957. DOI: 10.1111/rda.13734. View

Silyukova Y, Stanishevskaya O, Dementieva N . The current state of the problem of in vitro gene pool preservation in poultry. Vavilovskii Zhurnal Genet Selektsii. 2021; 24(2):176-184. PMC: 7716548. DOI: 10.18699/VJ20.611. View

Mehaisen G, Partyka A, Ligocka Z, Nizanski W . Cryoprotective effect of melatonin supplementation on post-thawed rooster sperm quality. Anim Reprod Sci. 2019; 212:106238. DOI: 10.1016/j.anireprosci.2019.106238. View

Partyka A, Nizanski W . Supplementation of Avian Semen Extenders with Antioxidants to Improve Semen Quality-Is It an Effective Strategy?. Antioxidants (Basel). 2021; 10(12). PMC: 8750219. DOI: 10.3390/antiox10121927. View

Riesco M, Robles V . Cryopreservation Causes Genetic and Epigenetic Changes in Zebrafish Genital Ridges. PLoS One. 2013; 8(6):e67614. PMC: 3689738. DOI: 10.1371/journal.pone.0067614. View

Partyka A, Lukaszewicz E, Nizanski W, Twardon J . Detection of lipid peroxidation in frozen-thawed avian spermatozoa using C(11)-BODIPY(581/591). Theriogenology. 2011; 75(9):1623-9. DOI: 10.1016/j.theriogenology.2011.01.002. View

Ard R, Maillet J, Daher E, Phan M, Zinoviev R, Parks R . PKCα-mediated phosphorylation of the diacylglycerol kinase ζ MARCKS domain switches cell migration modes by regulating interactions with Rac1 and RhoA. J Biol Chem. 2021; 296:100516. PMC: 8042443. DOI: 10.1016/j.jbc.2021.100516. View

Livak K, Schmittgen T . Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2002; 25(4):402-8. DOI: 10.1006/meth.2001.1262. View

10.

Izanloo H, Soleimanzadeh A, Bucak M, Imani M, Zhandi M . The effects of glutathione supplementation on post-thawed Turkey semen quality and oxidative stress parameters and fertilization, and hatching potential. Theriogenology. 2021; 179:32-38. DOI: 10.1016/j.theriogenology.2021.11.010. View

11.

Elliott R, Lloyd R, Fazeli A, Sostaric E, Georgiou A, Satake N . Effects of HSPA8, an evolutionarily conserved oviductal protein, on boar and bull spermatozoa. Reproduction. 2008; 137(2):191-203. DOI: 10.1530/REP-08-0298. View

12.

Qi X, Xing K, Huang Z, Chen Y, Wang L, Zhang L . Comparative transcriptome analysis digs out genes related to antifreeze between fresh and frozen-thawed rooster sperm. Poult Sci. 2020; 99(6):2841-2851. PMC: 7597663. DOI: 10.1016/j.psj.2020.01.022. View

13.

Chen W, Mao K, Liu Z, Dinh-Xuan A . The role of the RhoA/Rho kinase pathway in angiogenesis and its potential value in prostate cancer (Review). Oncol Lett. 2014; 8(5):1907-1911. PMC: 4186560. DOI: 10.3892/ol.2014.2471. View

14.

Wu H, Sun L, Wen Y, Liu Y, Yu J, Mao F . Major spliceosome defects cause male infertility and are associated with nonobstructive azoospermia in humans. Proc Natl Acad Sci U S A. 2016; 113(15):4134-9. PMC: 4839444. DOI: 10.1073/pnas.1513682113. View

15.

Mavi G, Dubey P, Cheema R . Association of antioxidant defense system with semen attributes vis a vis fertility in exotic and indigenous chicken breeds. Theriogenology. 2020; 144:158-163. DOI: 10.1016/j.theriogenology.2020.01.003. View

16.

Ansari M, Rakha B, Akhter S, Blesbois E, Santiago-Moreno J . Effect of Cryopreservation on Lipid Peroxidation, Antioxidant Potential, Chromatin Integrity, and Mitochondrial Activity of Indian Red Jungle Fowl () Semen. Biopreserv Biobank. 2019; 17(4):288-295. DOI: 10.1089/bio.2018.0111. View

17.

Holt W, Del Valle I, Fazeli A . Heat shock protein A8 stabilizes the bull sperm plasma membrane during cryopreservation: Effects of breed, protein concentration, and mode of use. Theriogenology. 2015; 84(5):693-701. DOI: 10.1016/j.theriogenology.2015.05.004. View

18.

Nishikimi M, Appaji N, Yagi K . The occurrence of superoxide anion in the reaction of reduced phenazine methosulfate and molecular oxygen. Biochem Biophys Res Commun. 1972; 46(2):849-54. DOI: 10.1016/s0006-291x(72)80218-3. View

19.

Garcia-Cardena G, Fan R, Shah V, Sorrentino R, Cirino G, Papapetropoulos A . Dynamic activation of endothelial nitric oxide synthase by Hsp90. Nature. 1998; 392(6678):821-4. DOI: 10.1038/33934. View

20.

Rakha B, Ansari M, Akhter S, Zafar Z, Naseer A, Hussain I . Dimethyleacetamide improves the cryosurvivability of Indian red jungle fowl (Gallus gallus murghi) sperm. Theriogenology. 2017; 103:83-89. DOI: 10.1016/j.theriogenology.2017.07.018. View