Circadian miR-449c-5p regulates uterine Ca2+ transport during eggshell calcification in chickens
Abstract Background miRNAs regulate circadian patterns by modulating the biological clocks of animals. In our previous study, we found that the clock gene exhibited a cosine expression pattern in the fallopian tube of chicken uterus. Clock-controlled miRNAs are present in mammals and Drosophila; how...
| Main Authors: | , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2021-10-01
|
| Series: | BMC Genomics |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12864-021-08074-3 |
| _version_ | 1818735929673646080 |
|---|---|
| author | Zhifu Cui Zhichao Zhang Felix Kwame Amevor Xiaxia Du Liang Li Yaofu Tian Xincheng Kang Gang Shu Qing Zhu Yan Wang Diyan Li Yao Zhang Xiaoling Zhao |
| author_facet | Zhifu Cui Zhichao Zhang Felix Kwame Amevor Xiaxia Du Liang Li Yaofu Tian Xincheng Kang Gang Shu Qing Zhu Yan Wang Diyan Li Yao Zhang Xiaoling Zhao |
| author_sort | Zhifu Cui |
| collection | DOAJ |
| description | Abstract Background miRNAs regulate circadian patterns by modulating the biological clocks of animals. In our previous study, we found that the clock gene exhibited a cosine expression pattern in the fallopian tube of chicken uterus. Clock-controlled miRNAs are present in mammals and Drosophila; however, whether there are clock-controlled miRNAs in the chicken uterus and, if so, how they regulate egg-laying rhythms is unclear. In this study, we selected 18 layer hens with similar ovipositional rhythmicity (each of three birds were sacrificed for study per 4 h throughout 24 h); their transcriptomes were scanned to identify the circadian miRNAs and to explore regulatory mechanisms within the uterus of chickens. Results We identified six circadian miRNAs that are mainly associated with several biological processes including ion trans-membrane transportation, response to calcium ion, and enrichment of calcium signaling pathways. Verification of the experimental results revealed that miR-449c-5p exhibited a cosine expression pattern in the chicken uterus. Ca2+-transporting ATPase 4 (ATP2B4) in the plasma membrane is the predicted target gene of circadian miR-449c-5p and is highly enriched in the calcium signaling pathway. We speculated that clock-controlled miR-449c-5p regulated Ca2+ transportation during eggshell calcification in the chicken uterus by targeting ATP2B4. ATP2B4 mRNA and protein were rhythmically expressed in the chicken uterus, and dual-luciferase reporter gene assays confirmed that ATP2B4 was directly targeted by miR-449c-5p. The expression of miR-449c-5p showed an opposite trend to that of ATP2B4 within a 24 h cycle in the chicken uterus; it inhibited mRNA and protein expression of ATP2B4 in the uterine tubular gland cells. In addition, overexpression of ATP2B4 significantly decreased intracellular Ca2+ concentration (P < 0.05), while knockdown of ATP2B4 accelerated intracellular Ca2+ concentrations. We found similar results after ATP2B4 knockdown by miR-449c-5p. Taken together, these results indicate that ATP2B4 promotes uterine Ca2+ trans-epithelial transport. Conclusions Clock-controlled miR-449c-5p regulates Ca2+ transport in the chicken uterus by targeting ATP2B4 during eggshell calcification. |
| first_indexed | 2024-12-18T00:29:04Z |
| format | Article |
| id | doaj.art-28599788874f441e8579f5268db09de7 |
| institution | Directory Open Access Journal |
| issn | 1471-2164 |
| language | English |
| last_indexed | 2024-12-18T00:29:04Z |
| publishDate | 2021-10-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Genomics |
| spelling | doaj.art-28599788874f441e8579f5268db09de72022-12-21T21:27:10ZengBMCBMC Genomics1471-21642021-10-0122111410.1186/s12864-021-08074-3Circadian miR-449c-5p regulates uterine Ca2+ transport during eggshell calcification in chickensZhifu Cui0Zhichao Zhang1Felix Kwame Amevor2Xiaxia Du3Liang Li4Yaofu Tian5Xincheng Kang6Gang Shu7Qing Zhu8Yan Wang9Diyan Li10Yao Zhang11Xiaoling Zhao12Department of Animal Science, Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, Sichuan Agricultural UniversityDepartment of Animal Science, Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, Sichuan Agricultural UniversityDepartment of Animal Science, Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, Sichuan Agricultural UniversityDepartment of Animal Science, Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, Sichuan Agricultural UniversityDepartment of Animal Science, Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, Sichuan Agricultural UniversityDepartment of Animal Science, Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, Sichuan Agricultural UniversityDepartment of Animal Science, Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, Sichuan Agricultural UniversityDepartment of Pharmacy, College of Veterinary Medicine, Sichuan Agricultural UniversityDepartment of Animal Science, Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, Sichuan Agricultural UniversityDepartment of Animal Science, Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, Sichuan Agricultural UniversityDepartment of Animal Science, Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, Sichuan Agricultural UniversityDepartment of Animal Science, Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, Sichuan Agricultural UniversityDepartment of Animal Science, Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, Sichuan Agricultural UniversityAbstract Background miRNAs regulate circadian patterns by modulating the biological clocks of animals. In our previous study, we found that the clock gene exhibited a cosine expression pattern in the fallopian tube of chicken uterus. Clock-controlled miRNAs are present in mammals and Drosophila; however, whether there are clock-controlled miRNAs in the chicken uterus and, if so, how they regulate egg-laying rhythms is unclear. In this study, we selected 18 layer hens with similar ovipositional rhythmicity (each of three birds were sacrificed for study per 4 h throughout 24 h); their transcriptomes were scanned to identify the circadian miRNAs and to explore regulatory mechanisms within the uterus of chickens. Results We identified six circadian miRNAs that are mainly associated with several biological processes including ion trans-membrane transportation, response to calcium ion, and enrichment of calcium signaling pathways. Verification of the experimental results revealed that miR-449c-5p exhibited a cosine expression pattern in the chicken uterus. Ca2+-transporting ATPase 4 (ATP2B4) in the plasma membrane is the predicted target gene of circadian miR-449c-5p and is highly enriched in the calcium signaling pathway. We speculated that clock-controlled miR-449c-5p regulated Ca2+ transportation during eggshell calcification in the chicken uterus by targeting ATP2B4. ATP2B4 mRNA and protein were rhythmically expressed in the chicken uterus, and dual-luciferase reporter gene assays confirmed that ATP2B4 was directly targeted by miR-449c-5p. The expression of miR-449c-5p showed an opposite trend to that of ATP2B4 within a 24 h cycle in the chicken uterus; it inhibited mRNA and protein expression of ATP2B4 in the uterine tubular gland cells. In addition, overexpression of ATP2B4 significantly decreased intracellular Ca2+ concentration (P < 0.05), while knockdown of ATP2B4 accelerated intracellular Ca2+ concentrations. We found similar results after ATP2B4 knockdown by miR-449c-5p. Taken together, these results indicate that ATP2B4 promotes uterine Ca2+ trans-epithelial transport. Conclusions Clock-controlled miR-449c-5p regulates Ca2+ transport in the chicken uterus by targeting ATP2B4 during eggshell calcification.https://doi.org/10.1186/s12864-021-08074-3Chicken uterineCircadian miRNAsTubular gland cellsCa2+ transport |
| spellingShingle | Zhifu Cui Zhichao Zhang Felix Kwame Amevor Xiaxia Du Liang Li Yaofu Tian Xincheng Kang Gang Shu Qing Zhu Yan Wang Diyan Li Yao Zhang Xiaoling Zhao Circadian miR-449c-5p regulates uterine Ca2+ transport during eggshell calcification in chickens BMC Genomics Chicken uterine Circadian miRNAs Tubular gland cells Ca2+ transport |
| title | Circadian miR-449c-5p regulates uterine Ca2+ transport during eggshell calcification in chickens |
| title_full | Circadian miR-449c-5p regulates uterine Ca2+ transport during eggshell calcification in chickens |
| title_fullStr | Circadian miR-449c-5p regulates uterine Ca2+ transport during eggshell calcification in chickens |
| title_full_unstemmed | Circadian miR-449c-5p regulates uterine Ca2+ transport during eggshell calcification in chickens |
| title_short | Circadian miR-449c-5p regulates uterine Ca2+ transport during eggshell calcification in chickens |
| title_sort | circadian mir 449c 5p regulates uterine ca2 transport during eggshell calcification in chickens |
| topic | Chicken uterine Circadian miRNAs Tubular gland cells Ca2+ transport |
| url | https://doi.org/10.1186/s12864-021-08074-3 |
| work_keys_str_mv | AT zhifucui circadianmir449c5pregulatesuterineca2transportduringeggshellcalcificationinchickens AT zhichaozhang circadianmir449c5pregulatesuterineca2transportduringeggshellcalcificationinchickens AT felixkwameamevor circadianmir449c5pregulatesuterineca2transportduringeggshellcalcificationinchickens AT xiaxiadu circadianmir449c5pregulatesuterineca2transportduringeggshellcalcificationinchickens AT liangli circadianmir449c5pregulatesuterineca2transportduringeggshellcalcificationinchickens AT yaofutian circadianmir449c5pregulatesuterineca2transportduringeggshellcalcificationinchickens AT xinchengkang circadianmir449c5pregulatesuterineca2transportduringeggshellcalcificationinchickens AT gangshu circadianmir449c5pregulatesuterineca2transportduringeggshellcalcificationinchickens AT qingzhu circadianmir449c5pregulatesuterineca2transportduringeggshellcalcificationinchickens AT yanwang circadianmir449c5pregulatesuterineca2transportduringeggshellcalcificationinchickens AT diyanli circadianmir449c5pregulatesuterineca2transportduringeggshellcalcificationinchickens AT yaozhang circadianmir449c5pregulatesuterineca2transportduringeggshellcalcificationinchickens AT xiaolingzhao circadianmir449c5pregulatesuterineca2transportduringeggshellcalcificationinchickens |