Asian-Australasian Journal of Animal Sciences

  • 제30권6호
  • /
  • Pages.781-787
  • /
  • 2017
  • /
  • 1011-2367(pISSN)
  • /
  • 1976-5517(eISSN)
아세아태평양축산학회 (Asian Australasian Association of Animal Production Societies)
권호 표지
DOI QR코드

DOI QR Code

Expression of Egr3 in mouse gonads and its localization and function in oocytes

  • Shin, Hyejin (Department of Biomedical Science and Technology, Institute of Biomedical Science and Technology, Konkuk University) ;
  • Seol, Dong-Won (Department of Biomedical Science, CHA University) ;
  • Nam, Minyeong (Department of Biomedical Science and Technology, Institute of Biomedical Science and Technology, Konkuk University) ;
  • Song, Haengseok (Department of Biomedical Science, CHA University) ;
  • Lee, Dong Ryul (Department of Biomedical Science, CHA University) ;
  • Lim, Hyunjung Jade (Department of Biomedical Science and Technology, Institute of Biomedical Science and Technology, Konkuk University)
  • 투고 : 2016.10.12
  • 심사 : 2016.12.07
  • 발행 : 2017.06.01
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

Objective: The early growth response (Egr) family consists of four members (Egr1, Egr2, Egr3, and Egr4) that are zinc finger transcription factors. Among them, Egr3 is involved in transcriptional regulation of target genes during muscle spindle formation and neurite outgrowth. We previously showed that the immunoreactive Egr3 is localized on oocyte spindle and accumulate near the microtubule organizing center during meiosis I in mice. Egr3 was also shown to be localized on spermatocytes. We herein investigated if Egr3 is expressed in mouse gonads and if Egr3 blockade results in any defect in oocyte maturation. Methods: Expression of Egr3 in mouse gonads was examined by reverse transcription-polymerase chain reaction. Full-length Egr3 and truncated Egr3 (${\Delta}Egr3$) complementary RNAs (cRNAs) with Xpress tag at N-terminus and DsRed2 at C-terminus, and small interfering RNA (siRNA) targeting Egr3 were microinjected into mouse oocytes at germinal vesicle stage. Localization of microinjected Egr3 was examined by confocal live imaging and immunofluorescence staining. Results: Egr3 mRNA was detected in mouse ovaries and testes from 1 to 4 week-old mice. An uncharacterized longer transcript containing 5'untranslated region was also detected in 3 and 4 week-old gonads. Microinjected Xpress-Egr3-DsRed2 or Xpress-${\Delta}Egr3$-DsRed2 localized to nuclei and chromosomes during meiotic progression. Microinjection of these cRNAs or Egr3 siRNA in oocytes did not affect meiotic maturation. Immunofluorescence staining of Egr3 in Xpress-${\Delta}Egr3$-DsRed2-injected oocytes showed a positive signal only on meiotic spindle, suggesting that this antibody does not detect endogenous or exogenous Egr3 in mouse oocytes. Conclusion: The results show that Egr3 localizes to chromosomes during meiotic progression and that certain antibodies may not faithfully represent localization of target proteins in oocytes. Egr3 seems to be dispensable during oocyte maturation in mice.

키워드

참고문헌

  1. O'Donovan KJ, Tourtellotte WG, Millbrandt J, Baraban JM. The EGR family of transcription-regulatory factors: progress at the interface of molecular and systems neuroscience. Trends Neurosci 1999;22: 167-73. https://doi.org/10.1016/S0166-2236(98)01343-5
  2. Shin H, Kwon S, Song H, Lim HJ. The transcription factor Egr3 is a putative component of the microtubule organizing center in mouse oocytes. PLoS One. 2014;9:e94708. https://doi.org/10.1371/journal.pone.0094708
  3. Lee SL, Sadovsky Y, Swirnoff AH, et al. Luteinizing hormone deficiency and female infertility in mice lacking the transcription factor NGFI-A (Egr-1). Science 1996;273:1219-21. https://doi.org/10.1126/science.273.5279.1219
  4. Kim HR, Kim YS, Yoon JA, et al. Egr1 is rapidly and transiently induced by estrogen and bisphenol A via activation of nuclear estrogen receptor-dependent ERK1/2 pathway in the uterus. Reprod Toxicol 2014;50:60-7. https://doi.org/10.1016/j.reprotox.2014.10.010
  5. Tourtellotte WG, Milbrandt J. Sensory ataxia and muscle spindle agenesis in mice lacking the transcription factor Egr3. Nat Genet 1998;20:87-91. https://doi.org/10.1038/1757
  6. Li L, Yun SH, Keblesh J, et al. Egr3, a synaptic activity regulated transcription factor that is essential for learning and memory. Mol Cell Neurosci 2007;35:76-88. https://doi.org/10.1016/j.mcn.2007.02.004
  7. Li S, Miao T, Sebastian M, et al. The transcription factors Egr2 and Egr3 are essential for the control of inflammation and antigen-induced proliferation of B and T cells. Immunity 2012;37:685-96. https://doi.org/10.1016/j.immuni.2012.08.001
  8. Fang F, Shangguan AJ, Kelly K, et al. Early growth response 3 (Egr-3) is induced by transforming growth factor-${\beta}$ and regulates fibrogenic responses. Am J Pathol. 2013;183:1197-208. https://doi.org/10.1016/j.ajpath.2013.06.016
  9. Quach DH, Oliveira-Fernandes M, Gruner KA, Tourtellotte WG. A sympathetic neuron autonomous role for Egr3-mediated gene regulation in dendrite morphogenesis and target tissue innervation. J Neurosci 2013;33:4570-83. https://doi.org/10.1523/JNEUROSCI.5481-12.2013
  10. Guerrero-Netro HM, Chorfi Y, Price CA. Effects of the mycotoxin deoxynivalenol on steroidogenesis and apoptosis in granulosa cells. Reproduction 2015;149:555-61. https://doi.org/10.1530/REP-15-0018
  11. Wang J, Zhao Y, Gu K, et al. The novel porcine gene early growth response 4 (Egr4) is differentially expressed in the ovaries of Erhualian and Pietrain pigs. Reprod Fertil Dev 2014;26:587-98. https://doi.org/10.1071/RD12380
  12. Hogarth CA, Mitchell D, Small C, Griswold M. EGR4 displays both a cell-and intracellular-specific localization pattern in the developing murine testis. Dev Dyn 2010;239:3106-14. https://doi.org/10.1002/dvdy.22442
  13. Mora GR, Olivier KR, Cheville JC, et al. The cytoskeleton differentially localizes the early growth response gene-1 protein in cancer and benign cells of the prostate. Mol Cancer Res 2004;2:115-28.
  14. Soubry A, Staes K, Parthoens E, et al. The transcriptional repressor Kaiso localizes at the mitotic spindle and is a constituent of the pericentriolar material. PLoS One 2010;5:e9203. https://doi.org/10.1371/journal.pone.0009203
  15. Wei H, Wang Q, Du J, et al. Unique subcellular distribution of RPB1 with a phosphorylated C-terminal domain (CTD) in mouse oocytes during meiotic division and its relationship with chromosome separation. J Reprod Dev 2015;61:541-8. https://doi.org/10.1262/jrd.2015-051
  16. Levkovitz Y, O'Donovan KJ, Baraban JM. Blockade of NGF-induced neurite outgrowth by a dominant-negative inhibitor of the egr family of transcription regulatory factors. J Neurosci 2001;21:45-52. https://doi.org/10.1523/JNEUROSCI.21-01-00045.2001
  17. O'Donovan KJ, Baraban JM. Major Egr3 isoforms are generated via alternate translation start sites and differ in their abilities to activate transcription. Mol Cell Biol 1999;19:4711-8. https://doi.org/10.1128/MCB.19.7.4711
  18. O'Donovan KJ, Levkovitz Y, Ahn D, Baraban JM. Functional comparison of Egr3 transcription factor isoforms: identification of an activation domain in the N-terminal segment absent from Egr3beta, a major isoform expressed in brain. J Neurochem 2000;75:1352-7.
  19. Baker M. Reproducibility crisis: Blame it on the antibodies. Nature 2015;521:274-6. https://doi.org/10.1038/521274a

피인용 문헌

  1. New markers for regulation of transcription and macromolecule metabolic process in porcine oocytes during in vitro maturation vol.21, pp.3, 2017, https://doi.org/10.3892/mmr.2020.10963
  2. Effect of genistein on the gene and protein expressions of CXCL–12 and EGR–1 in the rat ovary vol.105, pp.1, 2017, https://doi.org/10.1111/jpn.13448