Abstract
Broodiness is the primary factor influencing egg production in geese, in which several genes and miRNAs participate. Detailed spatiotemporal profiles of miRNAs encompassing follicle development levels, however, are lacking. In this study, we collected preovulatory follicles (classified as small white follicles, large white follicles, and small yellow follicles) from brooding and laying geese and aimed to analyze microRNA (miRNA or miR) during folliculogenesis. High-throughput sequencing and bioinformatics analysis were used to identify the miRNAs involved in follicle development. The let7 family, miR-10 family, and miR-143 family were abundant in these libraries, and they have been suggested to play a housekeeping role during folliculogenesis. Joint comparisons revealed 23 upregulated and 21 downregulated miRNAs (in at least two comparisons of follicles during brooding and laying, P < 0.1) in the laying stage. Unlike reproduction pathways reported for ovaries, GO and KEGG analysis suggested pathways for cell apoptosis and proliferation, such as the regulation of actin cytoskeleton, endocytosis, axon guidance, pathways in cancer, tight junctions, focal adhesion, the MAPK signaling pathway, cytokine-cytokine receptor interactions, and the Wnt signaling pathway in folliculogenesis. This study revealed the miRNAs that were directly involved in follicular atresia, and our results added to the understanding of the functional involvement of miRNAs during specific stages of follicle development.
Highlights
AVIANS HAVE THOUSANDS OF FOLLICLES in the ovary, and a strictly controlled follicular hierarchy is maintained, which is established through development, selection, and atresia in the follicles [24]
In this study, we obtained prehierarchal follicles at different levels during the brooding and laying stages [small white follicles (SWF) 2– 4 mm, large white follicles (LWF) 4 – 6 mm, and small yellow follicles (SYF, 6 – 8 mm); these follicles can be separated according to diameter and color, following the criteria described in Ref. 9], constructed small
We found that the miRNAs of the top 20 list are involved in regulating aromatase expression during follicle development, which is related to follicle development and survival, granulosa cell (GC) apoptosis, and ovarian cancer (Table 2)
Summary
AVIANS HAVE THOUSANDS OF FOLLICLES in the ovary, and a strictly controlled follicular hierarchy is maintained, which is established through development, selection, and atresia in the follicles [24]. After a single follicle is selected for final maturation and ovulation, there is little atresia associated with the unselected follicles within the avian prehierarchal cohort, which will be included for selection of a subsequent follicle ϳ24 h later [15] This well-organized follicular hierarchy controls the amount of egg production in birds until the ceasing period arrives, when the rest of the prehierarchal follicles atrophy. The homeostasis of selection and atresia in follicles is primarily orchestrated by synergistic interactions between extraovarian signals of hypothalamic-pituitary-gonadal origin and locally secreted growth factors. In this study, we obtained prehierarchal follicles at different levels during the brooding and laying stages [small white follicles (SWF) 2– 4 mm, large white follicles (LWF) 4 – 6 mm, and small yellow follicles (SYF, 6 – 8 mm); these follicles can be separated according to diameter and color, following the criteria described in Ref. 9], constructed small miRNA SEQUENCING IN PREHIERARCHAL FOLLICLES OF GEESE. The results may establish the spatiotemporal pattern of significant miRNAs in follicle development and reveal some useful information about follicle apoptosis in geese reproduction
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