Transcriptome Analysis Reveals miR-302a-3p Affects Granulosa Cell Proliferation by Targeting DRD1 in Chickens

Abstract

Egg production is an important economic trait in laying chickens as higher yields bring higher profits. Small yellow follicle (SYFL) development is a key determinant of chicken reproductive performance; however, the majority of SYFLs are not selected during the process of chicken reproduction and thus, atresia occurs. Although there have been numerous omic studies focused on egg production, the molecular mechanisms involved are still not well-understood. In this study, we used high-throughput technology to analyze the differences between the SYFL mRNA transcriptomes of high– (H) and low–egg-yielding (L) Taihang layer hens, with the aim of identifying the potential candidate genes involved in controlling the rate of egg production. We constructed six cDNA libraries, three from H and three from L Taihang hens and then performed high-throughput sequencing. Comparison of the H and L groups showed 415 differentially expressed genes (DEGs). In the high-yield group, 226 were upregulated and 189 were downregulated. Differentially enriched biological functions and processes were identified using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) database analysis. Ten of the candidate DEGs we identified (DRD1, MC5R, PCK1, CTSA, TGFBR3, AGO4, SLIT2, RGS1, SCNN1B, and ZP3) have been identified in previous studies as being involved in the development of small yellow follicles. DRD1 was significantly enriched in the gap junction pathway, which is an important pathway in chicken granulosa cells (GCs) to pass nutrition to an oocyte. Homology analysis showed that DRD1 was highly conserved in numerous species, indicating that it may be a productive target for improving egg production. Evidence from bioinformatics analysis revealed that gga-miR-302a-3p putatively targets the 3′UTR region of DRD1. We then identified the functions of gga-miR-302a-3p in follicular granulosa cell proliferation by targeting DRD1. RT-qPCR analysis showed that DRD1 and miR-302a-3p expression were inversely related in the SYLs of high and low egg-yielding chickens. Luciferase assays showed that miR-302a-3p targets the 3′UTR of DRD1, and overexpression of miR-302a-3p significantly inhibits the expression of DRD1 in chicken GCs (p < 0.01). Functional experiments revealed that by targeting DRD1, miR-302a-3p acts as an inhibitor of GC proliferation. Taken together, we concluded that miR-302a-3p affects chicken GC proliferation by targeting DRD1. Our data expanded the knowledge base of genes whose functions are important in egg production and the molecular mechanisms of high-yield egg production in chicken small yellow follicles.

Proteo-transcriptomic profiles reveal key regulatory pathways and functions of LDHA in the ovulation of domestic chickens (Gallus gallus)

“…Six DEGs, HSD3B1, LHCGR , NR5A1, CTSA, PTGS2, and RLN3, were selected to validate expression differences using qRT-PCR. We hypothesized that these six genes may be involved in regulating follicular development or ovulation process [33][34][35][36]. The qRT-PCR assay was conducted as previously described [37].…”

Interacting Networks of the Hypothalamic–Pituitary–Ovarian Axis Regulate Layer Hens Performance

“…As shown in Table 1. Chicken (hierarchal follicles) [113] miR-302a-3p DRD1 / Inhibits GCs proliferation Chicken (Small yellow follicle) [114] Abbreviations: Forkheadbox L2 (FOXL2); WNT family member 6 (Wnt6); forkhead box transcription factor O1 (FOXO1); B-cell lymphoma 1 (BCL1); trinucleotide repeat-containing gene 6a (TNRC6a); growth and differentiation factor 9 (GDF9); collagen type IV α 5 chain (COL4A5); insulin-like growth factor 2 mRNA-binding protein 3 (IGF2BP3); dopamine receptors 1 (DRD1); Krüppel-like factor 4 (KLF4); ATPase phospholipid transporting 8A1 (ATP8A1); complexin-1 (CPLX1); silencing information regulator 2 related enzyme 1 (SIRT1); mitogen-activated protein kinase 3 (MAPK3); microtubule associated protein RP/EB family member 1 (MAPRE1); catenin β 1 (CTNNB1); zine finger E-box binding homeobox (ZEB1); Smad interacting protein 1 (SIP1).…”

Long non-coding RNA MSTRG.5970.28 regulates proliferation and apoptosis of goose follicle granulosa cells via the miR-133a-3p/ANOS1 pathway

“…Follicular development is a key determinant of egg production performance in poultry ( Liu et al, 2022 ). Follicular development is a complex biological process with numerous genes and pathways involved in proliferation and differentiation ( Johnson et al, 2008 ).…”