Faheem, M. S., N. Ghanem, A. Gad, R. Procházka, and S. M. Dessouki, "Adaptive and Biological Responses of Buffalo Granulosa Cells Exposed to Heat Stress under In Vitro Condition.", Animals : an open access journal from MDPI, vol. 11, issue 3, 2021. Abstract

The steroidogenesis capacity and adaptive response of follicular granulosa cells (GCs) to heat stress were assessed together with the underlying regulating molecular mechanisms in Egyptian buffalo. In vitro cultured GCs were exposed to heat stress treatments at 39.5, 40.5, or 41.5 °C for the final 24 h of the culture period (7 days), while the control group was kept under normal conditions (37 °C). Comparable viability was observed between the control and heat-treated GCs at 39.5 and 40.5 °C. A higher release of E2, P4 and IGF-1 was observed in the 40.5 °C group compared with the 39.5 or 41.5 °C groups. The total antioxidant capacity was higher in response to heat stress at 39.5 °C. At 40.5 °C, a significant upregulation pattern was found in the expression of the stress resistance transcripts ( and ) and of . The relative abundance of was significantly downregulated for all heat-treated groups compared to the control, while was downregulated in GCs at 39.5 °C. Expression analyses of stress-related miRNAs (miR-1246, miR-181a and miR-27b) exhibited a significant downregulation in the 40.5 °C group compared to the control, whereas miR-708 was upregulated in the 39.5 and 40.5 °C groups. In conclusion, buffalo GCs exhibited different adaptive responses, to the different heat stress conditions. The integration mechanism between the molecular and secretory actions of the GCs cultured at 40.5 °C might provide possible insights into the biological mechanism through which buffalo GCs react to heat stress.

Ghanem, N., D. A. - E. R. Ahmed, S. M. Dessouki, M. S. Faheem, A. Y. Gad, J. Peippo, and A. H. Barkawi, "Cellular and molecular alterations of buffalo oocytes cultured under two different levels of oxygen tension during maturation.", Zygote (Cambridge, England), pp. 1-11, 2021. Abstract

This study was conducted to monitor the cellular and molecular changes of buffalo cumulus-oocytes complexes (COCs) cultured under high or low oxygen levels. Morphologically good quality COCs (n = 1627) were screened using brilliant cresyl blue (BCB) staining and placed into three groups (BCB+, BCB- and control). All groups of COCs were cultured under low (5%) or high (20%) oxygen tensions. Intracellular and molecular changes including oocyte ultrastructure, lipid contents, mitochondrial activity and transcript abundance of genes regulating different pathways were analyzed in the matured oocyte groups. The results revealed that oxygen tension did not affect cumulus expansion rates, however the BCB+ group had a higher (P ≤ 0.05) expansion rate compared with the BCB- group. BCB- oocytes recorded the lowest meiotic progression rate (P ≤ 0.05) under high oxygen levels that was linked with an increased level of reactive oxygen species (ROS) compared with the BCB+ oocytes. Ultrastructure examination indicated that BCB+ oocytes had a higher rate of cortical granules migration compared with BCB- under low oxygen tension. In parallel, our results indicated the upregulation of NFE2L2 in groups of oocytes cultured under high oxygen tension that was coupled with reduced mitochondrial activity. In contrast, the expression levels of MAPK14 and CPT2 genes were increased (P ≤ 0.05) in groups of oocytes cultured under low compared with high oxygen tension that was subsequently associated with increased mitochondrial activity. In conclusion, data from the present investigation indicated that low oxygen tension is a favourable condition for maintaining the mitochondrial activity required for nuclear maturation of buffalo oocytes. However, low-quality oocytes (BCB-) responded negatively to high oxygen tension by reducing the expression of gene-regulating metabolic activity (CPT2). This action was an attempt by BCB- oocytes to reduce the increased levels of endogenously produced ROS that was coupled with decreased expression of the gene controlling meiotic progression (MAPK14) in addition to nuclear maturation rate.

Faheem, M. S., S. M. Dessouki, F. E. S. Abdel-Rahman, and N. Ghanem, "Physiological and molecular aspects of heat-treated cultured granulosa cells of Egyptian buffalo (Bubalus bubalis).", Animal reproduction science, vol. 224, pp. 106665, 2021. Abstract

The physiological and molecular responses of granulosa cells (GCs) from buffalo follicles were investigated when there were in vitro heat stress conditions imposed. The cultured GCs were heat-treated at 40.5 °C for 24, 48 or 72 h while GCs of the control group were not heat-treated (37 °C). There were no differences in viability between control and heat-treated groups. There was an upward trend in increase in E secretion as the duration of heat stress advanced, being greater (P ≤ 0.05) for the GCs on which heat stress was imposed for 72 as compared with 24 h. In contrast, P release was less (P ≤ 0.05) from GCs heat-treated for 48 h than those cultured for 24 h and GCs of the control group. The relative abundance of ATP5F1A and SOD2 mRNA transcripts was consistent throughout the period when there was imposing of heat stress to sustain mitochondrial function. The relative abundance of CPT2 transcript was less in heat-treated GCs than in GCs of the control group. There was a greater relative abundance of SREBP1 and TNF-α mRNA transcripts after 48 h of heat-treatment of GCs than GCs of the control group. In conclusion, the results from the current study indicate buffalo GCs cultured when there was imposing of heat stress maintained normal viability, steroidogenesis and transcriptional profile. The stability of antioxidant status and increased transcription of genes regulating cholesterol biosynthesis and stress resistance may be defense mechanisms of buffalo GCs against heat stress.

Ahmed, D. A. - E. R., N. Ghanem, S. M. Dessouki, M. S. Faheem, A. Y. Gad, and A. H. Barkawi, "Developmental Competence of Buffalo Oocytes Cultured Under Different Oxygen Tensions after Selection with Brilliant Cresyl Blue", World’s Veterinary Journal, vol. 10, issue 2, pp. 246-253, 2020. wvj_102_246-253_june_25_2020.pdf
Ghanem, N., A. Amin, A. M. Saeed, S. M. Abdelhamid, A. El-Sayed, O. A. Farid, S. M. Dessouki, and M. S. Faheem, "Effects of Curcumin Supplementation on Viability and Antioxidant Capacity of Buffalo Granulosa Cells under In Vitro Culture Conditions", World’s Veterinary Journal, vol. 10, issue 2, pp. 146-159, 2020.
Dessouki, S. M., G. Ashour, M. El-Gayar, F. E. - H. El-Azzazi, E. Kodi, and N. Ghanem, "Gene Expression Profile and Enzymatic Activities of Frozen Buck Sperm Supplemented with Melatonin in Cold and Hot Temperatures", World’s Veterinary Journal, vol. 10, issue 1, pp. 125-136, 2020. wvj_101_125-136_march_25_2020_frozen_buck_sperm_activity_with_melatonin.pdf
Elsayed, D. H., F. E. El-Azzazi, Y. K. Mahmoud, S. M. Dessouki, and E. A. Ahmed, "Subclinical endometritis and postpartum ovarian resumption in respect to TNF-α, IL-8 and CRP in Egyptian buffaloes", Animal Reproduction, vol. 17, issue 1, pp. 1-13, 2020. crp.pdf
Abdelatty, A. M., O. A. M. Badr, S. A. Mohamed, M. S. Khattab, S. M. Dessouki, O. A. A. Farid, A. A. Elolimy, O. G. Sakr, M. A. El Hady, G. Mehesen, et al., "Long term conjugated linoleic acid supplementation modestly improved growth performance but induced testicular tissue apoptosis and reduced sperm quality in male rabbit", PLoS ONE, vol. 15, pp. 1-23, 2020.
Shakweer, W. M. E. - S., Y. M. Hafez, A. El-Sayed, S. M. Dessouki, I. M. Awadalla, and M. I. Mohamed, "Uptake of exogenous bovine GH–pmKate2– N expression vector by rams spermatozoa", Bulletin of the National Research Centre, vol. 43, issue 96, pp. 1-10, 2019. s42269-019-0136-4.pdf