Despite the widespread recognition of adaptive radiation as a driver of speciation, the mechanisms by which natural selection generates new species are incompletely understood. The evolutionary radiation of endemic East Asian cyprinids has been proposed as evolving through a change in spawning habits, involving a transition from semibuoyant eggs to adhesive eggs in response to crosslinked river-lake system formation. Here, we investigated the molecular mechanisms that underpin this radiation, associated with egg hydration and adhesiveness. We demonstrated that semibuoyant eggs enhance hydration by increasing the degradation of yolk protein and accumulation of Ca2+ and Mg2+ ions, while adhesive eggs improve adhesiveness and hardness of the egg envelope by producing an adhesive layer and a unique 4th layer to the egg envelope. Based on multiomics analyses and verification tests, we showed that during the process of adaptive radiation, adhesive eggs downregulated the "vitellogenin degradation pathway," "zinc metalloprotease pathway," and "ubiquitin-proteasome pathway" and the pathways of Ca2+ and Mg2+ active transport to reduce their hydration. At the same time, adhesive eggs upregulated the crosslinks of microfilament-associated proteins and adhesive-related proteins, the hardening-related proteins of the egg envelope, and the biosynthesis of glycosaminoglycan in the ovary to generate adhesiveness. These findings illustrate the novel molecular mechanisms associated with hydration and adhesiveness of freshwater fish eggs and identify critical molecular mechanisms involved in the adaptive radiation of endemic East Asian cyprinids. We propose that these key egg attributes may function as "magic traits" in this adaptive radiation.

Department of Ecology and Vertebrate Zoology University of Łódź Łódź Poland

Donghu Experimental Station of Lake Ecosystems State Key Laboratory of Freshwater Ecology and Biotechnology Institute of Hydrobiology Chinese Academy of Sciences Wuhan 430072 China

Institute of Ecological Research and Pollution Control of Plateau Lakes School of Ecology and Environment Yunnan University Kunming 650500 China

Institute of Vertebrate Biology Academy of Sciences of the Czech Republic Brno Czech Republic

University of Chinese Academy of Sciences Beijing 100049 China

Zobrazit více v PubMed

Darwin C. On the Origin of Species . John Murray; 1859.

Marques D. A., Meier J. I., Seehausen O. A combinatorial view on speciation and adaptive radiation. Trends in Ecology & Evolution . 2019;34(6):531–544. doi: 10.1016/j.tree.2019.02.008. PubMed DOI

McGee M. D., Borstein S. R., Meier J. I., et al. The ecological and genomic basis of explosive adaptive radiation. Nature . 2020;586(7827):75–79. doi: 10.1038/s41586-020-2652-7. PubMed DOI

Schluter D. The Ecology of Adaptive Radiation . Oxford: OUP; 2000.

Zhang Y., Teng D., Lu W., et al. A widely diverged locus involved in locomotor adaptation in Heliconius butterflies. Science Advances . 2021;7(32, article eabh2340) doi: 10.1126/sciadv.abh2340. PubMed DOI PMC

Lamichhaney S., Berglund J., Almén M. S., et al. Evolution of Darwin’s finches and their beaks revealed by genome sequencing. Nature . 2015;518(7539):371–375. doi: 10.1038/nature14181. PubMed DOI

Ronco F., Matschiner M., Böhne A., et al. Drivers and dynamics of a massive adaptive radiation in cichlid fishes. Nature . 2021;589(7840):76–81. doi: 10.1038/s41586-020-2930-4. PubMed DOI

Nosil P. Ecological Speciation . Oxford: OUP; 2012. DOI

Schluter D. Evidence for ecological speciation and its alternative. Science . 2009;323(5915):737–741. doi: 10.1126/science.1160006. PubMed DOI

Hendry A. P. Ecological speciation! Or the lack thereof?This perspective is based on the author’s J.C. Stevenson Memorial Lecture delivered at the Canadian Conference for Fisheries Research in Halifax, Nova Scotia, January 2008. Canadian Journal of Fisheries and Aquatic Sciences . 2009;66(8):1383–1398. doi: 10.1139/F09-074. DOI

Maan M. E., Seehausen O. Magic cues versus magic preferences in speciation. Evolutionary Ecology Research . 2012;14(6):779–785.

Servedio M. R., Van Doorn G. S., Kopp M., Frame A. M., Nosil P. Magic traits in speciation: ‘magic’ but not rare? Trends in Ecology & Evolution . 2011;26(8):389–397. doi: 10.1016/j.tree.2011.04.005. PubMed DOI

Servedio M. R. The relationship between sexual selection and speciation. Current Zoology . 2012;58(3):413–415. doi: 10.1093/czoolo/58.3.413. DOI

Eschmeyer W. N., Fricke R., Laan R. V. D. Eschmeyer’s catalog of fishes: genera, species, references. 2021. http://researcharchive.calacademy.org/research/ichthyology/catalog/fishcatmain.asp .

Chen F., Xue G., Wang Y., et al. Origin and evolution of the Yangtze River reconstructed from the largest molecular phylogeny of Cyprinidae. 2021. DOI

Chen F., Smith C., Wang Y., et al. The evolution of alternative buoyancy mechanisms in freshwater fish eggs. Frontiers in Ecology and Evolution . 2021;9 doi: 10.3389/fevo.2021.736718. DOI

Yu L., Lin J., Chen D., Duan X., Peng Q., Liu S. Ecological flow assessment to improve the spawning habitat for the four major species of carp of the Yangtze River: a study on habitat suitability based on ultrasonic telemetry. Water . 2018;10(5):p. 600. doi: 10.3390/w10050600. DOI

Kolar C. S., Chapman D. C., Courtenay W. R., Jr., Housel C. M., Williams J. D., Jennings D. P. Bigheaded carps: a biological synopsis and environmental risk assessment. American Fisheries Society Special Publication . 2007;33:1–204.

Cerdà J., Fabra M., Raldúa D. Physiological and molecular basis of fish oocyte hydration. In: Babin P. J., Cerdà J., Lubzens E., editors. The Fish Oocyte . Dordrecht: Springer; 2007. pp. 349–396. DOI

Finn R. N., Fyhn H. J. Requirement for amino acids in ontogeny of fish. Aquaculture Research . 2010;41(5):684–716. doi: 10.1111/j.1365-2109.2009.02220.x. DOI

Li H., Zhang S. Functions of vitellogenin in eggs. Oocytes . 2017;63:389–401. doi: 10.1007/978-3-319-60855-6_17. PubMed DOI

Sullivan C. V., Yilmaz O. Encyclopedia of Reproduction . Elsevier; 2018. Vitellogenesis and yolk proteins, fish; pp. 266–277. DOI

Cerdá J. L., Petrino T. R., Wallace R. A. Functional heterologous gap junctions in Fundulus ovarian follicles maintain meiotic arrest and permit hydration during oocyte maturation. Developmental Biology . 1993;160(1):228–235. doi: 10.1006/dbio.1993.1300. PubMed DOI

LaFleur G. J., Jr., Thomas P. Evidence for a role of Na+, K+-ATPase in the hydration of Atlantic croaker and spotted seatrout oocytes during final maturation. Journal of Experimental Zoology . 1991;258(1):126–136. doi: 10.1002/jez.1402580114. PubMed DOI

Wallace R. A., Greeley M. S., McPherson R. Analytical and experimental studies on the relationship between Na+, K+, and water uptake during volume increases associated with Fundulus oocyte maturation in vitro. Journal of Comparative Physiology B . 1992;162(3):241–248. doi: 10.1007/BF00357530. PubMed DOI

Rizzo E., Sato Y., Barreto B. P., Godinho H. P. Adhesiveness and surface patterns of eggs in neotropical freshwater teleosts. Journal of Fish Biology . 2002;61(3):615–632. doi: 10.1111/j.1095-8649.2002.tb00900.x. DOI

Riehl R., Patzner R. A. Minireview: the modes of egg attachment in teleost fishes. Italian Journal of Zoology . 1998;65(Supplement 1):415–420. doi: 10.1080/11250009809386857. DOI

Mansour N., Lahnsteiner F., Patzner R. A. Physiological and biochemical investigations on egg stickiness in common carp. Animal Reproduction Science . 2009;114(1–3):256–268. doi: 10.1016/j.anireprosci.2008.09.005. PubMed DOI

Niksirat H., Andersson L., Golpour A., Chupani L., James P. Quantification of egg proteome changes during fertilization in sterlet Acipenser ruthenus. Biochemical and Biophysical Research Communications . 2017;490(2):189–193. doi: 10.1016/j.bbrc.2017.06.019. PubMed DOI

Sano K., Kawaguchi M., Katano K., et al. Comparison of egg envelope thickness in teleosts and its relationship to the sites of ZP protein synthesis. Journal of Experimental Zoology Part B: Molecular and Developmental Evolution . 2017;328(3):240–258. doi: 10.1002/jez.b.22729. PubMed DOI

Wang Y., Chen F., He J., Xue G., Chen J., Xie P. Cellular and molecular modification of egg envelope hardening in fertilization. Biochimie . 2021;181:134–144. doi: 10.1016/j.biochi.2020.12.007. PubMed DOI

Yamagami K., Hamazaki T. S., Yasumasut S., Masuda K., luchi I. Molecular and cellular basis of formation, hardening, and breakdown of the egg envelope in fish. International Review of Cytology . 1992;136:51–92. doi: 10.1016/S0074-7696(08)62050-1. PubMed DOI

Hedrick J. L. Anuran and pig egg zona pellucida glycoproteins in fertilization and early development. International Journal of Developmental Biology . 2008;52(5–6):683–701. doi: 10.1387/ijdb.082580jh. PubMed DOI

Foster C. S., Thompson M. B., Van Dyke J. U., Brandley M. C., Whittington C. M. Emergence of an evolutionary innovation: gene expression differences associated with the transition between oviparity and viviparity. Molecular Ecology . 2020;29(7):1315–1327. doi: 10.1111/mec.15409. PubMed DOI

Gao W., Sun Y. B., Zhou W. W., et al. Genomic and transcriptomic investigations of the evolutionary transition from oviparity to viviparity. Proceedings of the National Academy of Sciences of the United States of America . 2019;116(9):3646–3655. doi: 10.1073/pnas.1816086116. PubMed DOI PMC

Qi P., Guo B., Xie C., et al. Assessing the genetic diversity and population structure of Culter alburnus in China based on mitochondrial 16S rRNA and COI gene sequences. Biochemical Systematics and Ecology . 2013;50:390–396. doi: 10.1016/j.bse.2013.04.010. DOI

Finn R. N., Østby G. C., Norberg B., Fyhn H. J. In vivo oocyte hydration in Atlantic halibut (Hippoglossus hippoglossus); proteolytic liberation of free amino acids, and ion transport, are driving forces for osmotic water influx. Journal of Experimental Biology . 2002;205(2):211–224. doi: 10.1242/jeb.205.2.211. PubMed DOI

Sørensen S. R., Butts I. A. E., Munk P., Tomkiewicz J. Effects of salinity and sea salt type on egg activation, fertilization, buoyancy and early embryology of European eel, Anguilla anguilla. Zygote . 2016;24(1):121–138. doi: 10.1017/S0967199414000811. PubMed DOI

Wootton R. J., Smith C. Reproductive Biology of Teleost Fishes . John Wiley & Sons; 2015.

Craik J. C. A., Harvey S. M. The causes of buoyancy in eggs of marine teleosts. Journal of the Marine Biological Association of the United Kingdom . 1987;67(1):169–182. doi: 10.1017/S0025315400026436. DOI

Thorsen A., Kjesbu O. S., Fyhndr H. J., Solemdal P. Physiological mechanisms of buoyancy in eggs from brackish water cod. Journal of Fish Biology . 1996;48(3):457–477. doi: 10.1111/j.1095-8649.1996.tb01440.x. DOI

Skoblina M. N. Hydration of oocytes in teleost fishes. Russian Journal of Developmental Biology . 2010;41(1):1–12. doi: 10.1134/S1062360410010017. DOI

Jia Y. D., Niu H. X., Meng Z., Liu X. F., Lei J. L. Biochemical composition of the ovarian fluid and its effects on the fertilization capacity of turbot Scophthalmus maximus during the spawning season. Journal of Fish Biology . 2015;86(5):1612–1620. doi: 10.1111/jfb.12676. PubMed DOI

Shoshan-Barmatz V., Gincel D. The voltage-dependent anion channel: characterization, modulation, and role in mitochondrial function in cell life and death. Cell Biochemistry and Biophysics . 2003;39(3):279–292. doi: 10.1385/CBB:39:3:279. PubMed DOI

Kwong J. Q., Molkentin J. D. Physiological and pathological roles of the mitochondrial permeability transition pore in the heart. Cell Metabolism . 2015;21(2):206–214. doi: 10.1016/j.cmet.2014.12.001. PubMed DOI PMC

Bernardi P., Rasola A., Forte M., Lippe G. The mitochondrial permeability transition pore: channel formation by F-ATP synthase, integration in signal transduction, and role in pathophysiology. Physiological Reviews . 2015;95(4):1111–1155. doi: 10.1152/physrev.00001.2015. PubMed DOI PMC

Fabra M., Raldúa D., Power D. M., Deen P. M., Cerda J. Marine fish egg hydration is aquaporin-mediated. Science . 2005;307(5709):545–545. doi: 10.1126/science.1106305. PubMed DOI

Chang Y. S., Wang Y. W., Huang F. L. Cross-linking of ZP2 and ZP3 by transglutaminase is required for the formation of the outer layer of fertilization envelope of carp egg. Molecular Reproduction and Development: Incorporating Gamete Research . 2002;63(2):237–244. doi: 10.1002/mrd.10174. PubMed DOI

Iuchi I., Ha C. R., Sugiyama H., Nomura K. Analysis of chorion hardening of eggs of rainbow trout, Oncorhynchus mykiss. Development, Growth & Differentiation . 1996;38(3):299–306. doi: 10.1046/j.1440-169X.1996.t01-2-00009.x. PubMed DOI

Welch M. D., Mullins R. D. Cellular control of actin nucleation. Annual Review of Cell and Developmental Biology . 2002;18(1):247–288. doi: 10.1146/annurev.cellbio.18.040202.112133. PubMed DOI

Pollard T. D., Borisy G. G. Cellular motility driven by assembly and disassembly of actin filaments. Cell . 2003;112(4):453–465. doi: 10.1016/S0092-8674(03)00120-X. PubMed DOI

Nuckolls G. H., Romer L. H., Burridge K. Microinjection of antibodies against talin inhibits the spreading and migration of fibroblasts. Journal of Cell Science . 1992;102(4):753–762. doi: 10.1242/jcs.102.4.753. PubMed DOI

Dos Remedios C. G., Chhabra D., Kekic M., et al. Actin binding proteins: regulation of cytoskeletal microfilaments. Physiological Reviews . 2003;83(2):433–473. doi: 10.1152/physrev.00026.2002. PubMed DOI

Cohen I., Young-Bandala L., Blankenberg T. A., Siefring G. E., Jr., Bruner-Lorand J. Fibrinoligase-catalyzed cross-linking of myosin from platelet and skeletal muscle. Archives of Biochemistry and Biophysics . 1979;192(1):100–111. doi: 10.1016/0003-9861(79)90075-4. PubMed DOI

Cohen I., Blankenberg T. A., Borden D., Kahn D. R., Veis A. Factor XIIIa-catalyzed cross-linking of platelet and muscle actin regulation by nucleotides. Biochimica et Biophysica Acta (BBA)-General Subjects . 1980;628(3):365–375. doi: 10.1016/0304-4165(80)90386-4. PubMed DOI

Mosher D. F., Schad P. E., Vann J. M. Cross-linking of collagen and fibronectin by factor XIIIa. Localization of participating glutaminyl residues to a tryptic fragment of fibronectin. Journal of Biological Chemistry . 1980;255(3):1181–1188. doi: 10.1016/S0021-9258(19)86160-4. PubMed DOI

Chang Y. S., Huang F. L. Fibroin-like substance is a major component of the outer layer of fertilization envelope via which carp egg adheres to the substratum. Molecular Reproduction and Development: Incorporating Gamete Research . 2002;62(3):397–406. doi: 10.1002/mrd.10125. PubMed DOI

Bravo Portela I., Martinez-Zorzano V. S., Molist-Perez I., Molist Garcia P. Ultrastructure and glycoconjugate pattern of the foot epithelium of the abalone Haliotis tuberculata (Linnaeus, 1758) (Gastropoda, Haliotidae) The Scientific World Journal . 2012;2012:12. doi: 10.1100/2012/960159.960159 PubMed DOI PMC

Högfors-Rönnholm E., Norrgård J., Wiklund T. Adhesion of smooth and rough phenotypes of Flavobacterium psychrophilum to polystyrene surfaces. Journal of Fish Diseases . 2015;38(5):429–437. doi: 10.1111/jfd.12250. PubMed DOI

Akimov S. S., Krylov D., Fleischman L. F., Belkin A. M. Tissue transglutaminase is an integrin-binding adhesion coreceptor for fibronectin. The Journal of Cell Biology . 2000;148(4):825–838. doi: 10.1083/jcb.148.4.825. PubMed DOI PMC

Akimov S. S., Belkin A. M. Cell surface tissue transglutaminase is involved in adhesion and migration of monocytic cells on fibronectin. Blood . 2001;98(5):1567–1576. doi: 10.1182/blood.V98.5.1567. PubMed DOI

Peramo A., Meads M. B., Dalton W. S., Matthews W. G. Static adhesion of cancer cells to glass surfaces coated with glycosaminoglycans. Colloids and Surfaces B: Biointerfaces . 2008;67(1):140–144. doi: 10.1016/j.colsurfb.2008.07.019. PubMed DOI

Selman K., Wallace R. A., Cerdà J. Bafilomycin A1 inhibits proteolytic cleavage and hydration but not yolk crystal disassembly or meiosis during maturation of sea bass oocytes. Journal of Experimental Zoology . 2001;290(3):265–278. doi: 10.1002/jez.1057. PubMed DOI

Zhang T., Rawson D. M., Tosti L., Carnevali O. Cathepsin activities and membrane integrity of zebrafish (Danio rerio) oocytes after freezing to −196°C using controlled slow cooling. Cryobiology . 2008;56(2):138–143. doi: 10.1016/j.cryobiol.2008.01.002. PubMed DOI

Robinson B. L., Dumas M., Ali S. F., Paule M. G., Gu Q., Kanungo J. Cyclosporine exacerbates ketamine toxicity in zebrafish: mechanistic studies on drug–drug interaction. Journal of Applied Toxicology . 2017;37(12):1438–1447. doi: 10.1002/jat.3488. PubMed DOI PMC

Lin Y. H., Hung G. Y., Wu L. C., Chen S. W., Lin L. Y., Horng J. L. Anion exchanger 1b in stereocilia is required for the functioning of mechanotransducer channels in lateral-line hair cells of zebrafish. PLoS One . 2015;10(2, article e0117041) doi: 10.1371/journal.pone.0117041. PubMed DOI PMC

Alternative signal pathways underly fertilization and egg activation in a fish with contrasting modes of spawning