RNA-Seq explores the functional role of the FGF10 gene in bovine adipocytes differentiation.

Objective: The present study was executed to explore the molecular mechanism of FGF10 gene in bovine adipogenesis. Methods: The bovine FGF10 gene was overexpressed through Ad-FGF10 or inhibited through siFGF10 and their NCs in bovine adipocytes, and the MOI, transfection efficiency, interference efficiency were evaluated through qRT-PCR, western blotting and fluorescence microscopy. The lipid droplets, TG content and the expression levels of adipogenic marker genes were measured during preadipocytes differentiation. The differentially expressed genes were explored through deep RNA sequencing. Results: The highest mRNA level was found in omasum, subcutaneous fat, and intramuscular fat. Moreover, the highest mRNA level was found in adipocytes at day 4 of differentiation. The results of red-oil o staining showed that overexpression (Ad-FGF10) of the FGF10 gene significantly (p<0.05) reduced the lipid droplets and TG content, and their down-regulation (siFGF10) increased the measurement of lipid droplets and TG in differentiated bovine adipocytes. Furthermore, the overexpression of the FGF10 gene down regulated the mRNA levels of adipogenic marker genes such as C/EBPa, FABP4, PPARy, LPL, and FAS, similarly, down-regulation of the FGF10 gene enriched the mRNA levels of C/EBPa, PPARy, FABP4, and LPL genes (P˂0.01). Additionally, the protein levels of PPARγ and FABP4 were reduced (p<0.05) in adipocytes infected with Ad-FGF10 gene and enriched in adipocytes transfected with siFGF10. Moreover, a total of 1774 DEGs (differentially expressed genes) including 157 up regulated and 1617 down regulated genes were explored in adipocytes infected with Ad-FGF10 or Ad-NC through deep RNA-sequencing. The top KEGG pathways regulated through DEGs were the PPAR signaling pathway, cell cycle, base excision repair, DNA replication, apoptosis, and regulation of lipolysis in adipocytes. Conclusion: Therefore, we can conclude that the FGF10 gene is a negative regulator of bovine adipogenesis and could be used as a candidate gene in marker-assisted selection.

In Vitro Fertilization in Kazakh Whiteheaded Cattle: A Comparative Study.

In vitro fertilization (IVF) technologies have great potential in the preservation of endangered species. In the current study, an IVF experiment was carried out to evaluate whether reproductive technologies are suitable for Kazakh Whiteheaded cattle, aimed at preserving this breed whose population has reduced drastically over the last thirty years. The reproduction characteristics of Kazakh Whiteheaded cows were compared to Aberdeen Angus cows. Transvaginal ultrasound-guided ovum pick up sessions were carried out followed by in vitro embryo production and embryo transfer and pregnancy diagnosis. The total and viable oocytes per OPU procedure were 12.8 ± 1.18 and 8.7 ± 0.85 for the Aberdeen Angus breed, and 8.8 ± 1.04 and 6.2 ± 0.83 for the Kazakh Whiteheaded breed. Similarly, the mean number of cleaved oocytes and morula/blastocyst stage embryos produced by OPU/IVF were 4.8 ± 0.49 and 1.4 ± 0.15 for the Aberdeen Angus breed, and 2.4 ± 0.46 and 0.18 ± 0.05 for the Kazakh Whiteheaded breed (p ≤ 0.02). From fifty Kazakh Whiteheaded donor animals, 2585 oocytes were aspirated following six ovum pick up sessions. One thousand eight hundred and seventy-six (72.5%) oocytes were chosen for maturation and were further fertilized. The number of embryos cleaved was 720 (38.3% out of oocytes fertilized) on day four post-fertilization. Of these cleaved embryos, 56 (7.5%) developed into the late morula/blastocyst stage on day seven post-fertilization, averaging 1.12 embryos per donor animal. Pregnancy was detected in 12 recipients; 4 healthy calves have been born to date. The outcomes of our study have demonstrated that reproductive technologies can be applicable in preserving the endangered Kazakh Whiteheaded cattle. The findings in this report will enhance knowledge of the reproductive characteristics of endangered domestic animals and help develop sophisticated reproductive protocols for animals with unique reproductive mechanisms.

The combined effects of temperature and relative humidity parameters on the reproduction of Stomoxys species in a laboratory setting.

In this study, Stomoxys species (S. calcitrans, S. sitiens and S. indica) were examined to improve on the current technique for mass rearing using a method of combined incubation parameters. Moreover, the reproductive potential of immature forms at various stages of development was defined. Immature forms of stable flies were incubated according to species. There was no significant difference in the number of immature forms obtained among species incubated under the same conditions. Six incubation parameters were used in combination, at temperatures (T) of 32°C, 27°C and 22°C and relative humidity (RH) of 90% and 70% RH. The combined method resulted in a higher number of eggs hatching at 32°C and 90% humidity as well as an increase in the number of larva pupated and emergence of imago at 27°C and 70% humidity.