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Homologues of p48 Protein from Morula Cells of Ascidian Styela rustica in Other Species of Stolidobranchia

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Abstract

Tunicata is an interesting phylogenetic group, at the base of the branch leading to Chordata. Ascidians (Tunicata, Ascidiacea) morula blood cells are involved in defense reactions and in the formation of the tunic extracellular matrix. The ascidian tunic is hardened as a result of sclerotization of matrix proteins due to the action of phenol oxidase enzyme, contained in morula cells. Morula cells of the ascidian Styela rustica contain two major proteins, the function of which is still unknown; one of them is a protein with molecular mass of 48 kDa – p48. The aim of present study was to search for possible homologues of p48 in ascidians belonging to the Stolidobranchia order: Styela coriacea (Styelidae), Molgula citrina (Molgulidae), Boltenia echinata, Halocynthia aurantium (Pyuridae) and to determine the tissue distribution of those homologues. In order to show the presence of p48 in the tissues of sea squirts we used indirect immunolabeling method on paraffin sections.It was shown that the antibodies bind with morula cells of Styela rustica, Styela coreacea, Boltenia echinata, with the tunic matrix in all studied species and with test cells of Styela rustica, Styela coreacea, Boltenia echinata, Molgula citrina. It gives us a ground to assume the existence of p48 homologues in all studied ascidians and to expect a common mechanism of their participation in the tunic formation, as possible substrates of the phenol oxidase system.

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ACKNOWLEDGMENTS

We are grateful to the staff of the Kartesh White Sea Biological Station (Zoological Institute, Russian Academy of Sciences) for the opportunity to carry out the work and for assistance in its implementation. We are also grateful to A.I. Solovyeva for assistance in collecting the material and L.S. Adonin for help in obtaining antibodies.

Funding

This work was supported by the Russian Foundation for Basic Research (project no. 15-04-06008), Russian Science Foundation (project no. 19-74-20102) and by the Program of the Presidium of Russian Academy of Sciences “Molecular and Cellular Biology” (no. 01.2.01457147).

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Authors and Affiliations

  1. Department of Cytology and Histology, St. Petersburg State University, 199034, St. Petersburg, Russia

    M. I. Tylets, O. I. Podgornaya & T. G. Shaposhnikova

  2. Institute of Cytology, Russian Academy of Sciences, 194064, St. Petersburg, Russia

    M. A. Daugavet & O. I. Podgornaya

  3. National Scientific Center of Marine Biology, Zhirmunsky Institute of Marine Biology, FEB RAS, 690041, Vladivostok, Russia

    A. V. Savelieva

  4. Far Eastern Federal University, 690922, Vladivostok, Russia

    O. I. Podgornaya

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  1. M. I. Tylets
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Correspondence to M. I. Tylets.

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Statement on the welfare of animals. The authors declare that all experiments on guinea pigs were carried out in compliance with animal rights and according to generally accepted international ethical standards guaranteed by a certificate of the Institute of Cytology, Russian Academy of Sciences, identification no. F18-00380 (Animal Welfare Assurance).

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Translated by A. Barkhash

 Abbreviations: DOPA—dioxyphenylalanine, BSA—bovine serum albumin, DAB—diaminobenzidine, SDS—sodium dodecyl sulfate, TBST (TBS-tween)—mixture of isotonic Tris-buffered saline solution and Tween 20 polysorbate.

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Tylets, M.I., Daugavet, M.A., Savelieva, A.V. et al. Homologues of p48 Protein from Morula Cells of Ascidian Styela rustica in Other Species of Stolidobranchia. Cell Tiss. Biol. 13, 388–396 (2019). https://doi.org/10.1134/S1990519X19050080

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