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Circadian miR-218-5p targets gene CA2 to regulate uterine carbonic anhydrase activity during egg shell calcification

Xiaxia Du1, Zhifu Cui1, Zifan Ning1

  • 1Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan Province, P. R. China; Key Laboratory of Livestock and Poultry Multi-omics, Ministry of Agricultural and Rural Affairs, College of Animal and Technology (Institute of Animal Genetics and Breeding), Sichuan Agricultural University, P. R., Chengdu, China.

Poultry Science
|September 27, 2022
PubMed

Insights

Circadian microRNA-218-5p regulates eggshell formation in chickens. It controls carbonic anhydrase activity by targeting the CA2 gene in the uterus, impacting eggshell quality.

Area of Science:

  • Reproductive Biology
  • Molecular Endocrinology
  • Chronobiology

Background:

  • MicroRNAs (miRNAs) are key regulators of the circadian clock.
  • miR-218-5p was previously identified as a circadian miRNA in the chicken uterus.
  • Its specific role in eggshell formation remained uncharacterized.

Purpose of the Study:

  • To elucidate the function of circadian miR-218-5p in chicken uterine eggshell formation.
  • To investigate the regulatory relationship between miR-218-5p, carbonic anhydrase 2 (CA2), and neuronal PAS domain protein 2 (NPAS2).
  • To determine the impact on carbonic anhydrase activity.

Main Methods:

  • Quantification of circadian expression of miR-218-5p, CA2, and NPAS2 in chicken uterus.
  • Dual-luciferase reporter gene assays to confirm direct targeting of CA2 and NPAS2 by miR-218-5p.
  • In vivo manipulation (overexpression and knockdown) of miR-218-5p and CA2.
  • Measurement of carbonic anhydrase activity.

Main Results:

  • miR-218-5p, CA2, and NPAS2 expression exhibited circadian oscillation in the chicken uterus.
  • miR-218-5p directly targets the 3' UTR of CA2 and NPAS2.
  • miR-218-5p inversely correlated with CA2 expression and regulated its mRNA and protein levels.
  • CA2 directly modulated carbonic anhydrase activity.
  • miR-218-5p influenced carbonic anhydrase activity by regulating CA2 expression.

Conclusions:

  • Clock-controlled miR-218-5p is a critical regulator of carbonic anhydrase activity in the chicken uterus.
  • This regulation occurs via targeting CA2 during the crucial process of eggshell formation.
  • Findings provide novel insights into the molecular mechanisms of avian eggshell development.
Keywords:
carbonic anhydrasechicken uteruscircadianeggshell formationmiR-218-5p

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