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. 2023 Jun 28;13(13):2132.
doi: 10.3390/ani13132132.

Transcriptome Profiling of Goose Ovarian Follicle Granulosa Cells Reveals Key Regulatory Networks for Follicle Selection

Jie Liu  1   2 , Shudi Dai  3 , Zichun Dai  1   2 , Yuyan Feng  4 , Mingming Lei  1   2 , Rong Chen  1   2 , Huanxi Zhu  1   2
Affiliations

Transcriptome Profiling of Goose Ovarian Follicle Granulosa Cells Reveals Key Regulatory Networks for Follicle Selection

Jie Liu et al. Animals (Basel). .

Abstract

The selection of follicles determines the reproductive performance of birds, but the process of follicle selection in geese is still elusive. This study focuses on Yangzhou geese during the egg-laying period and divides the follicular development process into three stages: small follicle development, follicle selection, and follicle maturation. Transcriptome sequencing was performed on granulosa cells from large white follicles, small yellow follicles, and F5 and F4 follicles. In addition, we selected the transcripts that remained unchanged during the development and maturation of small follicles but significantly changed during the follicular selection stage as the transcript collection that plays an important role in the follicular selection process. Then, we performed functional analysis on these transcripts and constructed a ceRNA network. The results showed that during the follicular selection stage, the number of differentially expressed mRNAs, miRNAs, and lncRNAs was the highest. In addition, miR-222-3p, miR-2954-3p, miR-126-5p, miR-2478, and miR-425-5p are potential key core regulatory molecules in the selection stage of goose follicles. These results can provide a reference for a better understanding of the basic mechanisms of the goose follicle selection process and potential targets for the precise regulation of goose egg production performance.

Keywords: ceRNA network; constructing; follicle selection; goose; granulosa cell; transcriptome profiling.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Schematic diagram of follicular development and sampling of Yangzhou geese ovarian follicles. The process of follicles from SWFs to ovulation can be divided into three stages: small follicular development, follicular selection, and follicular maturation. The follicle cohorts include pre-hierarchical (SWFs, LWFs, and SYFs) and hierarchical follicles (F5, F4, F3, F2, and F1). We collected LWFs, SYFs, and F5 and F4 follicles for experimental research. LWFs: the follicles are milky white with a diameter of 4–6 mm (blue arrow). SYFs: the follicles are milky yellow in color, with a diameter generally greater than 8 mm, and there are thin blood vessels on the surface of the follicles (pink arrow). F5: the follicles are orange in color, with a diameter greater than 12 mm, and numerous thick blood vessels distributed on the surface (yellow arrow). F4: the follicles are orange in color with a diameter greater than F5, and there are numerous thick blood vessels distributed on the surface (green arrow).
Figure 2
Figure 2
Principal component analysis of messenger RNAs (mRNAs) (A), microRNAs (miRNAs) (B), and long noncoding RNAs (lncRNAs) (C) transcriptome sequencing of granulosa cells. Follicles at the follicular development stage (LWFs to SYFs) or the follicular maturation stage (F5 to F4) are similar. The follicles in the follicular selection stage (SYFs to F5) have significant differences.
Figure 3
Figure 3
Number of upregulated or downregulated differentially expressed genes (DEGs) in granulosa cells: messenger RNAs (mRNAs) (A), microRNAs (miRNAs) (B), and long noncoding RNAs (lncRNAs) (C).
Figure 4
Figure 4
Time-series cluster analysis for the messenger RNAs (mRNAs), microRNAs (miRNAs), and long noncoding RNAs (lncRNAs) in the granulosa cells. Two expression patterns are displayed for mRNA (A,B), miRNA (C,D), and lncRNA (E,F).
Figure 5
Figure 5
Differentially expressed genes (DEGs) of granulosa cells stratified by messenger RNAs (mRNAs) (A), microRNAs (miRNAs) (B), and long noncoding RNAs (lncRNAs) (C) from three comparison groups.
Figure 6
Figure 6
Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways of DE-transcripts in granulosa cells: (A) top 20 KEGG pathways of DE-mRNAs; (B) top 20 KEGG pathways of DE-miRNAs; (C–E) top 20 KEGG pathways of antisense, cis, and trans lncRNAs.
Figure 7
Figure 7
A predicted ceRNA network based on miR-222-3p, miR-2954-3p, miR-126-5p, miR-2478, and miR-425-5p. Dark blue arrows, pink triangles, light blue circles, and green squares represent miRNA, mRNA, lncRNA, and circRNA, respectively.
Figure 8
Figure 8
Confirmation of messenger RNAs (mRNAs) (A–C), microRNAs (miRNAs) (D–F), and long noncoding RNAs (lncRNAs) (G–I) in granulosa cells by real-time PCR.
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