The mechanism of the effects of RBPJ on proliferation and gene expression in duck embryo fibroblasts

doi:10.3969/j.issn.1004-1524.20250649

Acta Agriculturae Zhejiangensis ›› 2026, Vol. 38 ›› Issue (7): 1336-1344.DOI: 10.3969/j.issn.1004-1524.20250649

• Animal Science • Previous Articles Next Articles

The mechanism of the effects of RBPJ on proliferation and gene expression in duck embryo fibroblasts

NIU Yuling^a(), ZHANG Yujie^a, JIA Luya^a, ZHAO Ayong^a, LIU Zhangguo^b, DU Xue^a^,^*()

^a College of Animal Science and Technology · College of Veterinary Medicine,Zhejiang A& F University,Hangzhou 311300,China
^b College of Food and Health (College of Modern Food Industry),Zhejiang A& F University,Hangzhou 311300,China

Received:2025-10-17 Online:2026-07-25 Published:2026-08-20

Abstract

Abstract:

During the egg-laying process, the oviduct of laying ducks must continuously undergo tissue remodeling, cell renewal, and homeostasis maintenance. Among these processes, fibroblasts, as the primary stromal cells, play a fundamental role in extracellular matrix remodeling, supporting epithelial differentiation, and maintaining smooth muscle function. The recombination signal binding protein for immunoglobulin kappa J region (RBPJ) is an important effector of the Notch signaling pathway, which regulates cell proliferation and differentiation, thereby affecting tissue development, morphology, and function. However, whether RBPJ regulates the function of duck oviduct by influencing duck oviduct fibroblasts has not yet been reported. This study investigated the effects of RBPJ on the viability and proliferation of duck embryo fibroblasts and its regulatory role in downstream gene expression through cell culture, RNA interference (RNAi), and transcriptome analysis. The results showed that: (1) Duck embryo fibroblasts were successfully isolated and RBPJ-interfered cell models were successfully constructed; (2) RBPJ gene interference significantly (p<0.05) reduced the viability at 12 h and the proliferation ability at 24 h of duck embryonic fibroblasts; (3) After RBPJ gene interference, 364 genes were differentially expressed and these significantly enriched in signaling pathways such as the MAPK and calcium signaling pathways. In conclusion: RBPJ affects the MAPK signaling pathway by regulating the expression of the PLA2G4C and AREG genes, and affects the calcium signaling pathway by regulating the expression of the FGF9, NTRK3, and AGTR1 genes, thereby affecting cell proliferation. These findings fill the research gap regarding the role of RBPJ in the proliferation of duck embryo fibroblasts. Moreover, they lay a research foundation for future analysis of RBPJ functions in duck oviduct fibroblasts and provide a theoretical basis and scientific support for improving egg-laying performance in practical production.

Key words: RBPJ gene, duck, oviduct, fibroblasts, cell function

CLC Number:

S834

NIU Yuling, ZHANG Yujie, JIA Luya, ZHAO Ayong, LIU Zhangguo, DU Xue. The mechanism of the effects of RBPJ on proliferation and gene expression in duck embryo fibroblasts[J]. Acta Agriculturae Zhejiangensis, 2026, 38(7): 1336-1344.

Figures/Tables 5

Table 1 siRNA sequences

siRNA名称 siRNA name	序列(5'→3') Sequence (5'→3')
NC-F	UUCUCCGAACGUGUCACGUTT
NC-R	ACGUGACACGUUCGGAGAATT
Si260-F	GCAGAAAUCAUAUGGAAAUTT
Si260-R	AUUUCCAUAUGAUUUCUGCTT
Si440-F	CUUGGAAGGAAAGAAUUAUTT
Si440-R	AUAAUUCUUUCCUUCCAAGTT

Table 2 Primer sequences

基因 Gene	正向引物序列(5'→3') Forward primer sequence (5'→3')	反向引物序列(5'→3') Reverse primer sequence (5'→3')	产物长度/bp Product length/bp
ACTB	AGCCATCTTTCTTGGGTATGGA	TTGTCACAAGGGTGTGGGTG	158
RBPJ	TAATCCAGTTTCAAGCCACTCC	CTTGGACTGGTTGACGGACC	165

Fig.1 Detection of RBPJ knockdown efficiency A, Relative mRNA expression level of RBPJ, normalized to ACTB; B, Representative Western blot bands of RBPJ protein; C, Relative gray value quantification of RBPJ protein expression, normalized to GAPDH. NC, negative control siRNA; Si260 and Si440, siRNAs targeting RBPJ. Different lowercase letters above the bars indicate significant differences (p<0.05); ns, No significant difference (p>0.05). The same applies to the following figures.

Fig.2 Effects of RBPJ siRNA on fibroblast proliferation and viability A, EdU assay was used to detect cell proliferation (scale bar=100 μm). Hoechst 33342 staining labeled the nuclei (blue), and EdU incorporation labeled S-phase cells (red), the Merge panels show the overlays of the two channels. B, Quantification of EdU-positive cell rates. C, Cell viability was measured by CCK-8 assay. * indicates a significant (p<0.05) difference between the NC group and the Si440 group.

Fig.3 The effect of RBPJ interference on fibroblast transcriptome profiles A, Principal component analysis; B, Volcano plot of differentially expressed genes; C, GO functional enrichment analysis of differentially expressed genes; D, KEGG pathway enrichment analysis of differentially expressed genes.

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The mechanism of the effects of RBPJ on proliferation and gene expression in duck embryo fibroblasts

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