Abstract
The Henze precipitate, a peculiar blue-green microparticulate obtained by lysis of the blood cells of the ascidian Phallusia mammillata (Protochordata), was investigated with atomic force microscopy (AFM), scanning electron microscopy (SEM) and X-ray microanalysis. The precipitate was collected from the Henze solution, an unstable red-brown product obtained by treating blood with distilled water, whose degradation yields a characteristic blue-green product. The microparticulates measured 50–100 µm in diameter and appeared irregular in shape. SEM examination showed smooth, roughly round boundaries. The microparticulate surface examined with AFM appeared as an irregular matrix formed by 70–320-nm-wide mammillate composites, including and embedding small (500–800 nm wide) crystal-like multilayered formations. X- ray analysis showed that the elements present in these same precipitates were mainly C, Si, Al and O. The microparticulate composition appeared close to those of natural waxes or lacquers, embedding amorphous silicates and/or other Si–Al components. The unusual occurrence of Si in ascidian blood and its role are discussed.
This is a preview of subscription content, log in via an institution to check access.
References
Binnig GH, Quate CF, Gerber C (1986) Atomic force microscopy. Phys Rev Lett 56:930–933
Butt HJ, Wolff EK, Gould SAC, Northern BD, Peterson CM, Hansma PK (1990) Imaging cells with the atomic force microscope. J Struct Biol 105:54–61
Chernoff EAG, Chernoff DA (1992) Atomic force microscope images of collagen fibers. J Vac Sci Technol A 10:596–599
Frank P, Carlson RMK, Hodgson KO (1988) Further investigations on the status of acidity and vanadium in the blood cells of Ascidia ceratodes. Inorg Chem 27:118–122
Frank P, Hedman B, Carlson RMK, Hodgson KO (1994) Interaction of vanadium and sulfate in blood cells from the tunicate Ascidia ceratodes: observations using X-ray absorption edge structure and EPR spectroscopies. Inorg Chem 33:3794–3803
Frank P, Kustin K, Robinson WE, Linebaugh L, Hodgson KO (1995) Nature and ligation of vanadium within whole blood cells and Henze solution from the tunicate Ascidia ceratodes, as investigated by using X-ray absorption spectrometry. Inorg Chem 34:5942–5949
Henderson E, Haydon PG, Sakaguchi DS (1992) Actin filaments dynamics in living glial cells imaged by atomic force microscopy. Science 257:1944–1946
Henze M (1911) Untersuchungen über das Blut der Ascidien. I. Hoppe Seyler’s Z Physiol Chem 72:494–501
Karrasch S, Hegerl R, Hoh J, Baumeister W, Engel A (1994) Atomic force microscopy produces faithful high-resolution images of protein surfaces in an aqueous environment. Proc Natl Acad Sci USA 91:836–838
Lindsay SM, Thundat T, Nagahara L, Knipping U, Rill RL (1989) Images of DNA double helix in water. Science 244:1063–1064
Michibata H, Hirata J, Uesaka M, Numakunai T, Sakurai H (1987) Separation of vanadocytes: determination and characterization of vanadium ion in the separated blood cells of the ascidian, Ascidia ahodori. J Exp Zool 244:33–38
Monniot F, Martoja R, Truchet M, Fröhlich F (1992) Opal in ascidian: a curious bioaccumulation in the ovary. Mar Biol 112:283–292
Muller DJ, Schoenenberger CA, Schabert F, Engel A (1997) Structural changes in native membrane proteins monitored at subnanometer resolution with the atomic force microscopy: a review. J Struct Biol 119:149–157
Nette G, Scippa S, Vincentiis M de (2000) Origin of the Henze solution/precipitate from morula cells of the blood of the ascidian Phallusia mammillata. Naturwissenschaften 87:220–224
Oltz EM, Bruening, RC, Smith, MJ, Kustin K, Nakanishi K (1988) The tunichromes: a class of reducing blood pigments from sea squirts: isolation, structures, and vanadium chemistry. J Am Chem Soc 110:6162–6172
Oltz EM, Pollack S, Delohery T, Smith MJ, Ojika M, Lee S, Kustin K, Nakanishi K (1989) Distribution of tunichrome and vanadium in sea squirt blood cells sorted by flow cytometry. Experientia 45:186–190
Ryan DE, Ghatlia ND, McDermott AE, Turro NJ, Nakanishi K, Kustin K (1992) Reactivity of tunichromes: reduction of vanadium(V) and vanadium(lV) to vanadium(lII) at neutral pH. J Am Chem Soc 114:9659–9660
Scippa S, Botte L, Zierold K, Vincentiis M de (1985) X-ray microanalytical studies on cryofixed blood cells of the ascidian Phallusia mammillata. I. Elemental composition of morula cells. Cell Tissue Res 239:459–461
Scippa S, Zierold K, Vincentiis M de (1988) X-ray microanalytical studies on cryofixed blood cells of the ascidian Phallusia mammillata. II. Elemental composition of the various blood cell types. J Submicrosc Cytol Pathol 20:719–730
Scippa S, Vincentiis M de, Zierold K (1990) Quantitative X-ray microanalysis of freeze-dried cryosections from the ascidian Phallusia mammillata. Int J Invert Reprod Dev 17:141–146
Taylor SW, Kammerer B, Bayer E (1997a) New perspectives in the chemistry and biochemistry of the tunichromes and related compounds. Chem Rev 97:333–346
Taylor SW, Kammerer B, Nicholson GJ, Pusacker K, Walk T, Bayer E, Scippa S, Vincentiis M de (1997b) Morulin Pm: a modified polypeptide containing TOPA and 6-bromotryptophan from the morula cells of the ascidian, Phallusia mammillata. Arch Biochem Biophys 348:278–288
Webb DA (1939) Observations on the blood of certain ascidians, with special reference to the biochemistry of vanadium. J Exp Biol 16:499–522
Acknowledgements
The authors gratefully acknowledge Prof. Ernst Bayer, Tubingen University, for his helpful suggestions; Dr. G. Grasso, CSATA Tecnopolis, Valenzano, Italy, for assistance with microanalysis; CNR, Italy for a grant (G.N.); Dr. Martin Wynne and peer reviewers for useful suggestions. Mr. Pasquale Sansone of Stazione Zoologica “A. Dohrn”, Napoli, is gratefully acknowledged for ascidian collection.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Ciancio, A., Scippa, S., Nette, G. et al. Analysis of the Henze precipitate from the blood cells of the ascidian Phallusia mammillata. Naturwissenschaften 91, 366–370 (2004). https://doi.org/10.1007/s00114-004-0540-x
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00114-004-0540-x