Dithiothreitol Affects the Fertilization Response in Immature and Maturing Starfish Oocytes
Abstract
1. Introduction
2. Materials and Methods
2.1. Gamete Collection, Oocyte Maturation, and Fertilization In Vitro
2.2. Light Microscopy and Transmission Electron Microscopy (TEM)
2.3. Chemicals and Reagents
2.4. Microinjection, Ca2+ Imaging, Fluorescent Labeling of F-Actin, and Jelly Coat
2.5. Visualization of Sperm inside Treated Oocytes before and after Maturation
2.6. Statistical Analysis
3. Results
3.1. The Effect of the Disulfide-Reducing Agent DTT on the Fertilization Response of A. aranciacus Immature Oocytes
3.2. DTT Treatment of Immature Starfish Oocytes Affects the Distribution of the Cortical F-Actin and Its Dynamics upon Insemination
3.3. DTT Treatment Does Not Induce Germinal Vesicle Breakdown (GVBD) and Maturation in A. aranciacus Starfish Oocytes
3.4. DTT Treatment Affects the F-Actin Rearrangement during Oocyte Maturation and Fertilization
3.5. Ultrastructural Changes in the Vitelline Layer and Microvillar Morphology Following DTT Treatment
3.6. Altered Sperm-Induced Ca2+ Response in the DTT-Treated Oocytes of Starfish
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Kanatani, H. Maturation-Inducing Substance in Starfishes. Int. Rev. Cytol. 1973, 35, 253–298. [Google Scholar] [CrossRef] [PubMed]
- Meijer, L.; Guerrier, P. Maturation and Fertilization in Starfish Oocytes. Int. Rev. Cytol. 1984, 86, 129–196. [Google Scholar] [CrossRef] [PubMed]
- Kishimoto, T. MPF-based meiotic cell cycle control: Half a century of lessons from starfish oocytes. Proc. Jpn. Acad. Ser. B 2018, 94, 180–203. [Google Scholar] [CrossRef] [PubMed]
- Chiba, K. Oocyte Maturation in Starfish. Cells 2020, 9, 476. [Google Scholar] [CrossRef]
- Dale, B.; de Santis, A.; Hoshi, M. Membrane response to 1-methyladenine requires the presence of the nucleus. Nature 1979, 282, 89–90. [Google Scholar] [CrossRef]
- Miyazaki, S.-I.; Hirai, S. Fast polyspermy block and activation potential: Correlated changes during oocyte maturation of a starfish. Dev. Biol. 1979, 70, 327–340. [Google Scholar] [CrossRef]
- Dale, B.; Dan-Sohkawa, M.; De Santis, A.; Hoshi, M. Fertilization of the starfish Astropecten aurantiacus. Exp. Cell Res. 1981, 132, 505–510. [Google Scholar] [CrossRef]
- Moody, W.J.; Bosma, M.M. Hormone-induced loss of surface membrane during maturation of starfish oocytes: Differential effects on potassium and calcium channels. Dev. Biol. 1985, 112, 396–404. [Google Scholar] [CrossRef]
- Schroeder, T.E.; Stricker, S.A. Morphological changes during maturation of starfish oocytes: Surface ultrastructure and cortical actin. Dev. Biol. 1983, 98, 373–384. [Google Scholar] [CrossRef]
- Longo, F.J.; Woerner, M.; Chiba, K.; Hoshi, M. Cortical changes in starfish (Asterina pectinifera) oocytes during 1-methyladenine-induced maturation and fertilisation/activation. Zygote 1995, 3, 225–239. [Google Scholar] [CrossRef]
- Barresi, M.J.F.; Gilbert, S.F. Beginning a New Organism. In Fertilization, 13th ed.; Oxford University Press: New York, NY, USA, 2024; pp. 211–246. [Google Scholar]
- Santella, L.; Chun, J.T. Structural actin dynamics during oocyte maturation and fertilization. Biochem. Biophys. Res. Commun. 2022, 633, 13–16. [Google Scholar] [CrossRef] [PubMed]
- Santella, L.; Limatola, N.; Chun, J.T. Cellular and molecular aspects of oocyte maturation and fertilization: A perspective from the actin cytoskeleton. Zool. Lett. 2020, 6, 5–21. [Google Scholar] [CrossRef] [PubMed]
- Santella, L.; Limatola, N.; Vasilev, F.; Chun, J.T. Maturation and fertilization of echinoderm eggs: Role of actin cytoskeleton dynamics. Biochem. Biophys. Res. Commun. 2018, 506, 361–371. [Google Scholar] [CrossRef] [PubMed]
- Mohri, T.; Kyozuka, K. Starfish oocytes of A. pectinifera reveal marked differences in sperm-induced electrical and intracellular calcium changes during oocyte maturation and at fertilization. Mol. Reprod. Dev. 2022, 89, 3–22. [Google Scholar] [CrossRef]
- Limatola, N.; Chun, J.T.; Schneider, S.C.; Schmitt, J.-L.; Lehn, J.-M.; Santella, L. The Effect of Acidic and Alkaline Seawater on the F-Actin-Dependent Ca2+ Signals Following Insemination of Immature Starfish Oocytes and Mature Eggs. Cells 2023, 12, 740. [Google Scholar] [CrossRef]
- Just, E.E. The Production of Filaments by Echinoderm Ova as a Response to Insemination, with Special Reference to the Phenomenon as Exhibited by Ova of the Genus Asterias. Biol. Bull. 1929, 57, 311–325. [Google Scholar] [CrossRef]
- Dan, J.C. Studies on the acrosome. II Acrosome reaction in starfish spermatozoa. Biol. Bull. 1954, 107, 203–218. [Google Scholar] [CrossRef]
- Hoshi, M.; Moriyama, H.; Matsumoto, M. Structure of acrosome reaction-inducing substance in the jelly coat of starfish eggs: A mini review. Biochem. Biophys. Res. Commun. 2012, 425, 595–598. [Google Scholar] [CrossRef]
- Clark, J.M. An Experimental Study of Polyspermy. Biol. Bull. 1936, 70, 361–384. [Google Scholar] [CrossRef]
- Just, E.E. The Biology of the Cell Surface; P. Blakiston's Son & Co., Inc.: Philadelphia, PA, USA, 1939. [Google Scholar]
- Fujimori, T.; Hirai, S. Differences in starfish oocyte susceptibility to polyspermy during the course of maturation. Biol. Bull. 1979, 157, 249–257. [Google Scholar] [CrossRef]
- Santella, L.; Limatola, N.; Chun, J.T. Actin Cytoskeleton and Fertilization in Starfish Eggs. In Sexual Reproduction in Animals and Plants—Part II: Gametogenesis, Gamete Recognition, Activation, and Evolution; Sawada, H., Inoue, N., Iwano, M., Eds.; Springer Open: Tokyo, Japan, 2014; pp. 141–155. [Google Scholar] [CrossRef]
- Mabuchi, I. Purification from starfish eggs of a protein that depolymerizes actin. J. Biochem. 1981, 89, 1341–1344. [Google Scholar] [CrossRef]
- Mabuchi, I. An actin-depolymerizing protein (Depactin) from starfish oocytes: Properties and interaction with actin. J. Cell Biol. 1983, 97, 1612–1621. [Google Scholar] [CrossRef]
- Otto, J.J.; Schroeder, T.E. Assembly-disassembly of actin bundles in starfish oocytes: An analysis of actin-associated proteins in the isolated cortex. Dev. Biol. 1984, 101, 263–273. [Google Scholar] [CrossRef]
- Chun, J.T.; Santella, L. Roles of the actin-binding proteins in intracellular Ca2+ signalling. Acta Physiol. 2009, 195, 61–70. [Google Scholar] [CrossRef] [PubMed]
- Limatola, N.; Vasilev, F.; Chun, J.T.; Santella, L. Altered actin cytoskeleton in ageing eggs of starfish affects fertilization process. Exp. Cell Res. 2019, 381, 179–190. [Google Scholar] [CrossRef] [PubMed]
- Kishimoto, T.; Kanatani, H. Induction of starfish oocyte maturation by disulfide-reducing agents. Exp. Cell Res. 1973, 82, 296–302. [Google Scholar] [CrossRef]
- Kishimoto, T.; Cayer, M.L.; Kanatani, H. Starfish oocyte maturation and reduction of disulfide-bond on oocyte surface. Exp. Cell Res. 1976, 101, 104–110. [Google Scholar] [CrossRef] [PubMed]
- Nemoto, S.-I. Nature of the 1-methyladenine-requiring phase in maturation of starfidh oocytes. Develop. Growth Differ. 1982, 24, 429–442. [Google Scholar] [CrossRef]
- Mita, M.; Ueta, N.; Nagahama, Y. In Vitro Induction of Starfish Oocyte Maturation by Cysteine Alkylesters: (oocyte maturation/starfish/1-MeAde/SH-group). Dev. Growth Differ. 1987, 29, 607–616. [Google Scholar] [CrossRef]
- Mita, M. Incapacity of Response to Disulfide-Reducing Agent in Triton X-100–Treated Oocytes of Starfish. Asterina Pectinifera Ann. N. Y. Acad. Sci. 2005, 1040, 413–416. [Google Scholar] [CrossRef]
- Epel, D.; Weaver, A.M.; Mazia, D. Methods for removal of the vitelline membrane of sea urchin eggs. Exp. Cell Res. 1970, 61, 64–68. [Google Scholar] [CrossRef] [PubMed]
- Eddy, E.M.; Shapiro, B.M. Changes in the topography of the sea urchin egg after fertilization. J. Cell Biol. 1976, 71, 35–48. [Google Scholar] [CrossRef]
- Thaler, C.D.; Kuo, R.C.; Patton, C.; Preston, C.M.; Yagisawa, H.; Epel, D. Phosphoinositide metabolism at fertilization of sea urchin eggs measured with a GFP-probe. Dev. Growth Differ. 2004, 46, 413–423. [Google Scholar] [CrossRef] [PubMed]
- Limatola, N.; Chun, J.T.; Cherraben, S.; Schmitt, J.-L.; Lehn, J.-M.; Santella, L. Effects of Dithiothreitol on Fertilization and Early Development in Sea Urchin. Cells 2021, 10, 3573. [Google Scholar] [CrossRef] [PubMed]
- Limatola, N.; Chun, J.T.; Santella, L. Species-Specific Gamete Interaction during Sea Urchin Fertilization: Roles of the Egg Jelly and Vitelline Layer. Cells 2022, 11, 2984. [Google Scholar] [CrossRef] [PubMed]
- Vacquier, V.D.; Moy, G.W. Isolation of bindin: The protein responsible for adhesion of sperm to sea urchin eggs. Proc. Natl. Acad. Sci. USA 1977, 74, 2456–2460. [Google Scholar] [CrossRef] [PubMed]
- Vacquier, V.D. The quest for the sea urchin egg receptor for sperm. Biochem. Biophys. Res. Commun. 2012, 425, 583–587. [Google Scholar] [CrossRef]
- Wessel, G.M.; Wada, Y.; Yajima, M.; Kiyomoto, M. Sperm lacking Bindin are infertile but are otherwise indistinguishable from wildtype sperm. Sci. Rep. 2021, 11, 21583. [Google Scholar] [CrossRef]
- Wessel, G.M.; Wada, Y.; Yajima, M.; Kiyomoto, M. Bindin is essential for fertilization in the sea urchin. Proc. Natl. Acad. Sci. USA 2021, 118, e2109636118. [Google Scholar] [CrossRef]
- Limatola, N.; Chun, J.T.; Santella, L. Regulation of the Actin Cytoskeleton-Linked Ca2+ Signaling by Intracellular pH in Fertilized Eggs of Sea Urchin. Cells 2022, 11, 1496. [Google Scholar] [CrossRef]
- Shen, S.S.; Steinhardt, R.A. Direct measurement of intracellular pH during metabolic depression of the sea urchin egg. Nature 1978, 272, 253–254. [Google Scholar] [CrossRef] [PubMed]
- Begg, D.A.; Rebhun, L.I. pH regulates the polymerization of actin in the sea urchin egg cortex. J. Cell Biol. 1979, 83, 241–248. [Google Scholar] [CrossRef] [PubMed]
- Tilney, L.G.; Jaffe, L.A. Actin, microvilli, and the fertilization cone of sea urchin eggs. J. Cell Biol. 1980, 87, 771–782. [Google Scholar] [CrossRef] [PubMed]
- Carron, C.P.; Longo, F. Relation of cytoplasmic alkalinization to microvillar elongation and microfilament formation in the sea urchin egg. Dev. Biol. 1982, 89, 128–137. [Google Scholar] [CrossRef] [PubMed]
- Mabuchi, I.; Hosoya, H.; Sakai, H. Actin in the cortical layer of the sea urchin egg. Changes in its content during and after fertilization. Biomed. Res. 1980, 1, 417–426. [Google Scholar] [CrossRef]
- Yonemura, S.; Mabuchi, I. Wave of cortical actin polymerization in the sea urchin egg. Cell Motil. Cytoskelet. 1987, 7, 46–53. [Google Scholar] [CrossRef]
- Chun, J.T.; Puppo, A.; Vasilev, F.; Gragnaniello, G.; Garante, E.; Santella, L. The Biphasic Increase of PIP2 in the Fertilized Eggs of Starfish: New Roles in Actin Polymerization and Ca2+ Signaling. PLoS ONE 2010, 5, e14100. [Google Scholar] [CrossRef][Green Version]
- Santella, L.; Lim, D.; Moccia, F. Calcium and fertilization: The beginning of life. Trends Biochem. Sci. 2004, 29, 400–408. [Google Scholar] [CrossRef] [PubMed]
- Santella, L.; Chun, J.T. Actin, more than just a housekeeping protein at the scene of fertilization. Sci. China Life Sci. 2011, 54, 733–743. [Google Scholar] [CrossRef][Green Version]
- Santella, L.; Vasilev, F.; Chun, J.T. Fertilization in echinoderms. Biochem. Biophys. Res. Commun. 2012, 425, 588–594. [Google Scholar] [CrossRef]
- Santella, L.; Limatola, N.; Chun, J.T. Calcium and actin in the saga of awakening oocytes. Biochem. Biophys. Res. Commun. 2015, 460, 104–113. [Google Scholar] [CrossRef] [PubMed]
- Miyazaki, S.H.; Ohmori, H.; Sasaki, S. Action potential and non-linear current voltage relation in starfish oocytes. J. Physiol. 1975, 246, 37–54. [Google Scholar] [CrossRef] [PubMed]
- Santella, L.; De Riso, L.; Gragnaniello, G.; Kyozuka, K. Cortical granule translocation during maturation of starfish oocytes requires cytoskeletal rearrangement triggered by InsP3-mediated Ca2+ release . Exp. Cell Res. 1999, 248, 567–574. [Google Scholar] [CrossRef] [PubMed]
- Shôji, Y.; Hamaguchi, M.S.; Hiramoto, Y. Mechanical properties of the endoplasm in starfish oocytes. Exp. Cell Res. 1978, 117, 79–87. [Google Scholar] [CrossRef]
- Nemoto, S.-I.; Yoneda, M.; Uemura, I. Marked decrease in the rigidity of starfish oocytes induced by 1-methyladenine. Develop. Growth Differ. 1980, 22, 315–325. [Google Scholar] [CrossRef]
- Newman, S.A. Just’s “independent irritability” revisited: The activated egg as excitable soft matter. Mol. Reprod. Dev. 2009, 76, 966–974. [Google Scholar] [CrossRef]
- Byrnes, W.M.; Newman, S.A. Ernest Everett Just: Egg and Embryo as Excitable Systems. J. Exp. Zool. B Mol. Dev. Evol. 2014, 322, 191–201. [Google Scholar] [CrossRef]
- Lange, K. Microvillar Ca++ signaling: A new view of an old problem. J. Cell. Physiol. 1999, 180, 19–34. [Google Scholar] [CrossRef]
- Lange, K. Microvillar Ion Channels: Cytoskeletal Modulation of Ion Fluxes. J. Theor. Biol. 2000, 206, 561–584. [Google Scholar] [CrossRef]
- Gartzke, J.; Lange, K. Cellular target of weak magnetic fields: Ionic conduction along actin filaments of microvilli. Am. J. Physiol.-Cell Physiol. 2002, 283, C1333–C1565. [Google Scholar] [CrossRef]
- Vasilev, F.; Limatola, N.; Chun, J.T.; Santella, L. Contributions of suboolemmal acidic vesicles and microvilli to the intracellular Ca2+ increase in the sea urchin eggs at fertilization. Int. J. Biol. Sci. 2019, 15, 757–775. [Google Scholar] [CrossRef] [PubMed]
- Ishiwata, S. Freezing of actin. Reversible oxidation of a sulfhydryl group and structural change. J. Biochem. 1976, 80, 595–609. [Google Scholar] [CrossRef] [PubMed]
- Tang, J.X.; Janmey, P.A.; Stosselm, T.P.; Ito, T. Thiol Oxidation of Actin Produces Dimers That Enhance the Elasticity of the F-Actin Network. Biophys. J. 1999, 76, 2208–2215. [Google Scholar] [CrossRef]
- Hosoda, E.; Hiraoka, D.; Hirohashi, N.; Omi, S.; Kishimoto, T.; Chiba, K. SGK regulates pH increase and cyclin B–Cdk1 activation to resume meiosis in starfish ovarian oocytes. J. Cell Biol. 2019, 218, 3612–3629. [Google Scholar] [CrossRef]
- Chiba, K.; Kado, R.T.; Jaffe, L.A. Development of calcium release mechanisms during starfish oocyte maturation. Dev. Biol. 1990, 140, 300–306. [Google Scholar] [CrossRef]
- Spudich, A.; Wrenn, J.T.; Wessells, N.K. Unfertilized sea urchin eggs contain a discrete cortical shell of actin that is subdivided into two organizational states. Cell Motil. Cytoskelet. 1988, 9, 85–96. [Google Scholar] [CrossRef]
- Henson, J.H.; Begg, D.A. Filamentous actin organization in the unfertilized sea urchin egg cortex. Dev. Biol. 1988, 127, 338–348. [Google Scholar] [CrossRef]
- Gillot, I.; Ciapa, B.; Payan, P.; Sardet, C. The calcium content of cortical granules and the loss of calcium from sea urchin eggs at fertilization. Dev. Biol. 1991, 146, 396–405. [Google Scholar] [CrossRef] [PubMed]
- Chun, J.T.; Limatola, N.; Vasilev, F.; Santella, L. Early events of fertilization in sea urchin eggs are sensitive to actin-binding organic molecules. Biochem. Biophys. Res. Commun. 2014, 450, 1166–1174. [Google Scholar] [CrossRef]
- Cline, D.J.; Redding, S.E.; Brohawn, S.G.; Psathas, J.N.; Schneider, J.P.; Thorpe, C. New water-soluble phosphines as reductants of peptide and protein disulfide bonds: Reactivity and membrane permeability. Biochemistry 2004, 43, 15195–15203. [Google Scholar] [CrossRef] [PubMed]
- Vasilev, F.; Chun, J.T.; Gragnaniello, G.; Garante, E.; Santella, L. Effects of ionomycin on egg activation and early development in starfish. PLoS ONE 2012, 7, e39231. [Google Scholar] [CrossRef] [PubMed]
- Lim, D.; Lange, K.; Santella, L. Activation of oocytes by latrunculin A. FASEB J. 2002, 16, 1050–1056. [Google Scholar] [CrossRef] [PubMed]
- Just, E.E. Initiation of development in Arbacia. IV. Some cortical reactions as criteria for optimum fertilization capacity and their significance for the physiology of development. Protoplasma 1928, 5, 97–126. [Google Scholar] [CrossRef]
GV-Stage Oocytes (N = 3) | SW 10 min | DTT 10 min | DTT 10 min, Wash |
---|---|---|---|
Ca2+ response | 16 out of 16 | 13 out of 16 | 11 out of 13 |
CF | 12 out of 16 | 12 out of 16 | 10 out of 13 |
CF (RFU) | 0.166 ± 0.010 | 0.047 ± 0.07 | 0.103 ± 0.04 |
Ca2+ wave (RFU) | 0.59 ± 0.011 | 0.31 ± 0.09 * | 0.26 ± 0.04 * |
Traverse time (s) | 134.04 ± 3.87 | 125.44 ± 7.1 | 132.68 ± 6.75 |
Time of activation (s) | 19.6 ± 4.6 | 16.43 ± 2.4 | 19.9 ± 5.6 |
N spots | 4.56 ± 0.87 | 3.07 ± 0.6 * | 2.63 ± 0.75 * |
Maturing Oocytes (N = 5) | 1-MA 70 min | DTT 70 min | 1-MA 60 min + DTT 10 min | 1-MA 60 min + DTT 10 min, Wash |
---|---|---|---|---|
Ca2+ response | 38 out of 38 | 9 out of 18 | 24 out of 25 | 17 out of 18 |
CF | 32 out of 38 | 1 out of 18 | 18 out of 25 | 8 out of 18 |
CF (RFU) | 0.095 ± 0.03 | 0.036 | 0.031 ± 0.01 * | 0.063 ± 0.02 # |
Ca2+ wave (RFU) | 1.19 ± 0.13 | 0.89 ± 0.25 * | 0.55 ± 0.19 * | 0.70 ± 0.09 * |
Traverse time (s) | 129.9 ± 12.05 | 134.2 ± 16.23 | 117.7 ± 9.8 | 129.4 ± 5.6 |
Time of activation (s) | 34.15 ± 8.9 | 203.45 ± 97.6 * | 61.91 ± 35.03 | 35.62 ± 11.67 |
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Limatola, N.; Chun, J.T.; Chiba, K.; Santella, L. Dithiothreitol Affects the Fertilization Response in Immature and Maturing Starfish Oocytes. Biomolecules 2023, 13, 1659. https://doi.org/10.3390/biom13111659
Limatola N, Chun JT, Chiba K, Santella L. Dithiothreitol Affects the Fertilization Response in Immature and Maturing Starfish Oocytes. Biomolecules. 2023; 13(11):1659. https://doi.org/10.3390/biom13111659
Chicago/Turabian StyleLimatola, Nunzia, Jong Tai Chun, Kazuyoshi Chiba, and Luigia Santella. 2023. "Dithiothreitol Affects the Fertilization Response in Immature and Maturing Starfish Oocytes" Biomolecules 13, no. 11: 1659. https://doi.org/10.3390/biom13111659
APA StyleLimatola, N., Chun, J. T., Chiba, K., & Santella, L. (2023). Dithiothreitol Affects the Fertilization Response in Immature and Maturing Starfish Oocytes. Biomolecules, 13(11), 1659. https://doi.org/10.3390/biom13111659