Profiling several key milk miRNAs and analysing their signalling pathways in dairy sheep breeds during peak and late lactation.

BACKGROUND: The comprehensive understanding of microRNAs (miRNAs) in sheep milk during various lactation periods and their impact on milk yield and composition remains limited. OBJECTIVES: This study aimed to investigate the expression patterns of four highly expressed miRNAs in sheep milk and their association with milk composition and yield parameters during peak and late lactation stages. METHODS: A total of 40 healthy 4-year-old Akkaraman (n = 20) and Awassi (n = 20) ewes registered with the Ministry of Agriculture and Forestry of the Republic of Türkiye were used in the present study. For miRNA isolation from milk, the Qiagen miRNeasy Serum/Plasma Advanced Kit was utilised following the manufacturer's instructions. The expression levels of miRNAs were assessed using Qiagen miRNA PCR Assays. RESULTS: The significant fold changes in the expression levels of oar-miR-30a-5p, oar-miR-148a and oar-miR-181a were observed between peak and late lactation periods in the Awassi sheep breed. Conversely, only oar-miR-30a-5p and oar-miR-148a exhibited statistically significant changes in the Akkaraman sheep breed during the same lactation periods. Furthermore, oar-miR-21-5p demonstrated a significant fold change exclusively in peak lactation compared to Akkaraman and Awassi ewes. CONCLUSIONS: The findings suggest that the expression of the analysed miRNAs is influenced by both the lactation stage and different sheep breeds. This study offers valuable insights into the relationship between key miRNA expressions in sheep milk and milk composition and yield parameters during peak and late lactation, contributing to the existing knowledge in this field.

Genetic analysis of PALB2 gene WD40 domain in canine mammary tumour patients.

BACKGROUND: DNA repair mechanisms are essential for tumorigenesis and disruption of HR mechanism is an important predisposing factor of human breast cancers (BC). PALB2 is an important part of the HR. There are similarities between canine mammary tumours (CMT) and BCs. As its human counterpart, PALB2 mutations could be a predisposing factor of CMT. OBJECTIVES: In this study, we aimed to investigate the impacts of PALB2 variants on tumorigenesis and canine mammary tumor (CMT) malignancy. METHODS: We performed Sanger sequencing to detect germline mutations in the WD40 domain of the canine PALB2 gene in CMT patients. We conducted in silico analysis to investigate the variants, and compared the germline PALB2 mutations in humans that cause breast cancer (BC) with the variants detected in dogs with CMT. RESULTS: We identified an intronic (c.3096+8C>G) variant, two exonic (p.A1050V and p.R1354R) variants, and a 3' UTR variant (c.4071T>C). Of these, p.R1354R and c.4071T>C novel variants were identified for the first time in this study. We found that the p.A1050V mutation had a significant effect. However, we could not determine sufficient similarity due to the differences in nucleotide/amino acid sequences between two species. Nonetheless, possible variants of human sequences in the exact location as their dog counterparts are associated with several cancer types, implying that the variants could be crucial for tumorigenesis in dogs. Our results did not show any effect of the variants on tumor malignancy. CONCLUSIONS: The current project is the first study investigating the relationship between the PALB2 gene WD40 domain and CMTs. Our findings will contribute to a better understanding of the pathogenic mechanism of the PALB2 gene in CMTs. In humans, variant positions in canines have been linked to cancer-related phenotypes such as familial BC, endometrial tumor, and hereditary cancer predisposition syndrome. The results of bioinformatics analyses should be investigated through functional tests or case-control studies.

Transcriptome analysis and potential mechanisms of bovine oocytes under seasonal heat stress.

Heat stress is the major factor affecting cattle fertility but molecular mechanisms of deleterious impacts of elevated temperature on oocyte are still not well known. Therefore, the aim of this study is to gain a better understanding of the underlying molecular mechanism of how heat stress affects GV-stage and MII-stage oocytes and discover hub genes to heat resistance for cow oocytes. In this study, we used the bioinformatics approach to discover the differentially expressed genes between GV-stage and MII-stage oocytes, which were collected during spring and summer. When GV-stage oocytes were compared to MII-stage oocytes collected in July (Jul DEGs group) a total of 1068 genes were found as differentially expressed as a result of heat stress. Also, HSPA8, COPS5, POLR2L, PSMC6, and TPI1 were identified as the common top ranked genes for the Jul DEGs group. The highest connected hub gene for the Jul DEGs group was determined as HSPA8. Our results showed that different heat response mechanisms might be activated to protect oocytes from elevated temperatures in cattle. The identified genes and their associated pathways might play an important role in the response to heat stress that affects the oocytes in cattle.

Oar-miR-16b and oar-miR-27a: negatively correlated with milk yield and milk protein in sheep.

The characterization of miRNAs from sheep milk and their effect on milk yield and composition in sheep are remain unclear. Therefore, the aim of this study was to determine the expression pattern of several important miRNAs, which are associated with lactation in the sheep milk between high and low lactating-yield ewe groups. In addition to experimentally obtained miRNA expression results, the miRNA target genes were determined by bioinformatics analysis to identify biological pathways involved. miRNAs found to differ significantly in the expression level between the groups were oar-miR-181a, oar-miR-23a, oar-miR-27a, oar-miR-16b and oar-miR-374. Also, oar-miR-27a was shown negative correlation with milk protein and lactose contents while oar-miR-16b was shown negative correlation with milk yield in the high milk yield group. The highest connected hub genes for miR-27a target genes were determined as MAPK14 and PPARG. Also, six genes (HSPA4L, DNAJA2, ATP6V1B2, PPP2R1A, PPP2R1B, and PRKAR2A) were detected as hub genes for miR-16b. In this study, the relationship between expression profiles of several important miRNAs in sheep milk and milk yield and milk composition were investigated for the first time in high and low lactating yield groups.

Affecting Lipid Metabolism Salivary MicroRNAs Expressions in Arabian Racehorses Before and After the Race.

The active roles of microribonucleic acids (miRNAs) in gene regulation have made miRNAs a key point for the scientific world in the study of physiological processes. Although saliva includes the largest number of miRNAs, there is no miRNA study in saliva on horses has been found. Our study is the first study on miRNAs isolation from saliva in horses. In the present study, saliva was studied in Arabian racehorses to better understand the molecular mechanisms of expression levels that are effective in lipid metabolism of miRNAs and their target genes during the race. Identification of lipid metabolism of miRNAs and their target genes is an opportunity to provide information about biomarkers in Arabian racehorses on energy supply for race performance. Arabian racehorses have low glycogen content and high triglyceride storage capability, thanks to the high amount of oxidative type I fiber in their muscle tissue. Therefore, Arabian racehorses can provide higher levels of energy using more fat. The aim of this study is to determine the prerace and postrace expression levels of eight miRNAs in saliva that are known to affect lipid metabolism in Arabian racehorses. The expression level of eca-miR-33a was found to be statistically significant (P < .05). Target genes of eca-miR-33a have been copredicted as ABCA1, CROT, ABHD2, and SATB2, with three validated databases and other analysis tools. In conclusion, these findings revealed that both eca-miR-33a and its target genes could be potential core genes that play important roles in lipid metabolism in Arabian racehorses to provide energy during the race.