Giant basal spicule from the deep-sea glass sponge <italic>Monorhaphis chuni</italic>: synthesis of the largest bio-silica structure on Earth by silicatein

doi:10.1007/s11706-009-0044-x

Front Mater Sci Chin

2009, Vol. 3

Issue (3) : 226-240 https://doi.org/10.1007/s11706-009-0044-x

RESEARCH ARTICLE

Giant basal spicule from the deep-sea glass sponge Monorhaphis chuni: synthesis of the largest bio-silica structure on Earth by silicatein

Xiao-hong WANG¹(

), Xue-hua ZHANG², Heinz C. SCHR?DER³, Werner E. G. MüLLER³(

)

1. National Research Center for Geoanalysis, 26 Baiwanzhuang Dajie, Beijing 100037,China; 2. Laboratory of Guangzhou Marine Geological Surey, Guangzhou 510760, China; 3. Institut für Physiologische Chemie, Abteilung Angewandte Molekularbiologie, Universit?t, Duesbergweg 6, D-55099 Mainz, Germany

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Abstract

Like all sponges (phylum Porifera), the glass sponges (Hexactinellida) are provided with an elaborate and distinct body plan, which relies on a filigree skeleton. It is constructed by an array of morphologically determined elements, the spicules. Schulze described the largest siliceous hexactinellid sponge on Earth, the up to 3 m high Monorhaphis chuni, collected during the German Deep Sea Expedition “Valdivia” (1898–1899). This species develops an equally large bio-silica structure, the giant basal spicule (3 m × 10 mm). Using these spicules as a model, one can obtain the basic knowledge on the morphology, formation, and development of silica skeletal elements. The silica matrix is composed of almost pure silica, endowing it with unusual optophysical properties, which are superior to those of man-made waveguides. Experiments suggest that the spicules function in vivo as a nonocular photoreception system. The spicules are also provided with exceptional mechanical properties. Like demosponges, the hexactinellids synthesize their silica enzymatically via the enzyme silicatein (27 kDa protein). This enzyme is located in/embedded in the silica layers. This knowledge will surely contribute to a further utilization and exploration of silica in biomaterial/biomedical science.

Keywords sponge Porifera Hexactinellida spicule giant basal spicule silicatein biomaterial science

Corresponding Author(s): WANG Xiao-hong,Email:wxh0408@hotmail.com (Prof. X.H. WANG); MüLLER Werner E. G.,Email:wmueller@uni-mainz.de (Prof. W.E.G. MüLLER)

Issue Date: 05 September 2009

Cite this article:

Xiao-hong WANG,Xue-hua ZHANG,Heinz C. SCHR?DER, et al. Giant basal spicule from the deep-sea glass sponge Monorhaphis chuni: synthesis of the largest bio-silica structure on Earth by silicatein[J]. Front Mater Sci Chin, 2009, 3(3): 226-240.

URL:

https://academic.hep.com.cn/foms/EN/10.1007/s11706-009-0044-x
https://academic.hep.com.cn/foms/EN/Y2009/V3/I3/226

Fig.1 specimens. Young specimens are anchored to the muddy substratum by one single giant basal spicule (gbs). The body (bo) surrounds the spicule as a continuous and round cylinder. Schematic representation of the growth phases of the sessile animal with its giant basal spicule (gbs) that anchors the animal to the substratum and holds its surrounding soft body (bo). The characteristic habitus displays linearly arranged large atrial openings (at) of approximately 2 cm in diameter. With growth, the soft body dies off in the basal region and exposes the bare giant basal spicule (a to c). Part of the body (bo) with its atrial openings (at). The body surface is interspersed with ingestion openings allowing a continuous water flow though canals in the interior, which opens into oscules that are centralized in atrial openings, the sieve-plates. Original photograph of an specimen, collected during the Valdivia expedition. A dried specimen of a “moderate” size of 120 cm showing that the basal body had fallen off from the giant basal spicule (gbs). The apical body comprises the serially arranged atrial openings (at). A 2.7 m long giant basal spicule. The diameter of a giant basal spicule at the basis or in the middle of the spicule. in its natural soft bottom habitat of bathyal slopes of New Caledonia []. ((A)(C)(D): modified after Schulze []; (I): modified after Müller et al. [])

Fig.2 Structural architecture of the giant basal spicules from . Cross break through a giant basal spicule. The zoning into the lamellar region (la), the axial cylinder (cy), and the axial canal (ac) is seen. Lamellae (la) from a giant basal spicule that have been spalled off for the spicule releasing the axial cylinder. Structural arrangement of a giant basal spicule; cross section with the lamellae (la), the axial cylinder (cy), and the axial canal (ac). Axial cylinder with its axial canal (ac) and its axial filament (af). Axial filament released from a giant basal spicule. Presentation of the organic matrix within the lamellae after limited exposure to HF (hydrofluoric acid). At this magnification, the proteinaceous palisade-like scaffold (pr) within the lamellae (la) is uncovered. ((A)(B): digital light microscopy using a VHX-600 Digital Microscope from KEYENCE, Neu-Isenburg; Germany; (C)(D)(E)(F): high resolution scanning electron microscopy (HR-SEM))

Fig.3 Light optical properties and chemical analysis of the giant basal spicule. Original spicule, collected 1898/1899 at a depth of 1079 m off the coast of Somalia (Museum für Naturkunde Berlin, Germany; ZMB Por 12700) was used as an optical waveguide. The silica spicule is an excellent waveguide, while the calcareous material of the corals (co) is nontransparent. Electron microprobe analysis of a giant basal spicule. (C) Polished cross section through the spicule (sp) showing the lamellar-wise organization (la) at the outer region. The spicule is surrounded by fused polyps of the coral (co); SEM analysis. X-ray maps of elements ((B) and (D)) of the area shown in (C). The counts of elemental characteristic X-ray increase, so the concentrations of the elements increase from blue via green to yellow and, finally, to red. Elemental mapping is given for the elements Si (C) and Ca (E).

Fig.4 Toward the elucidation of the control mechanisms involved in the morphogenesis of the giant basal spicules. Scheme summarizing the three morphological zones of the spicules. The silica structures (with their organic components); the axial canal (ac) (axial filament (af)), the axial cylinder (cy) (axial barrel), and the lamellar zone (la) (lamellar coating). The surface of the spicule is surrounded by a net (net) that leaves open 10 μm holes, allowing the bio-silica lamellae to be squeezed in. Backcoupling mechanism explaining the interaction between the bio-silica formation caused by the silicatein in the axial filament (af) and the morphogenetic activity, displayed by the product of silicatein, the bio-silica. (B-a) At first, the axial filament (af) is formed, which mediates the formation of bio-silica (B-b). (B-c) Bio-silica of the formed lamellae (i) attracts the cells to associate with the growing spicule and (ii) causes a morphogenetic effect on the stem cells to differentiate into the sclerocytes (bio-silica-forming cells; sc) and the collencytes (collagen-forming cells; co). These two cell types allow the composition of the fibrous net (net) around the spicules. Self-assembly of silicatein; SEM analysis. The lamellae of had been extracted, and the resulting 25/27 kDa protein was analyzed for the ability to form organizes into a filamentous structures; SEM analysis. Stepwise dissolution of the silica shell around the axial filament of the spicule (comitalia) by treatment with HF solution; light microscopic analysis. Initial phase of the dissolution of the silica by HF. Solubilization of the lamellar zone (la) starts after 1–3 min, leaving behind proteinaceous filaments/clumps. The axial cylinder (cy) can be distinguished within the spicule by Nomarsky imaging. After complete dissolution of the lamellar zone (treatment for 30 min), the axial cylinder (cy) that surrounds the axial canal/axial filament (af) is exposed; stained positive with Sirius Red.

Fig.5 Collagen sheet formed around the giant spicules (sp) of . Early description of the fibrous sheath (fs) around the spicules by Schulze []. MicroCT analysis of a giant basal spicule. Three-dimensional reconstruction to resolve a cross section through a spicule in a noninvasive way. The section shows the organic envelope (env) that tightly surrounds the inorganic spicule (sp). The outer surface of the spicule shows a serrated relief structure (ss). Smaller spicules are surrounded by a collagen (col) net that is regularly punctured with holes (h) through which the underlying siliceous spicular material is visible. Axial section through a spicule discloses “rectangular” protrusions of the same size as the holes in the collagen net seen in (C). The dimensions of these protrusions fit into the space uncovered by the holes within the collagen sheet (>h<). Lamellae (la) form the protrusions and follow the curvings. ((C)(D): modified after Müller et al. [])

Fig.6 Analysis of the proteins, extracted from the giant basal spicules of . Analysis of the total spicule extract by two-dimensional gel electrophoresis (first, isoelectric focusing and, then, size separation). The arrows mark the positions of the two sets of proteins in the total spicule extract, the 27/30 kDa molecules and the 70 kDa polypeptides. The gel was stained with Coomassie Brilliant Blue; according to Müller et al. []. Corresponding Western blot to the protein extract obtained from the lamellae. Analysis of a total spicule extract by two-dimensional gel electrophoresis (first, isoelectric focusing and, then, size separation). The arrows mark the positions of the two sets of proteins in the total spicule extract, the 27/30 kDa molecules, and the 70 kDa polypeptides. The gel was stained with Coomassie Brilliant Blue.

Fig.7 Kinetic parameters, characterizing the proteolytic activity existing in extracts from the lamellae of giant basal spicules (the 25 kDa protein) using the substrate Z-Phe-Arg-AMC (the incubation period was 10 min (22°C)). Determination of the initial reaction velocity between 2 and 12 μmol/L of the substrate. Lineweaver-Burk plot applied for the determination of the hydrolytic activity of the protein extract from lamellae. Various concentrations of substrate (Z-Phe-Arg-AMC) were added to the standard assay. A double reciprocal plot of velocity substrate concentration was computed. After incubation, the degree of fluorescence of the released AMC was determined and used for the calculation of the enzymatic parameters; the values are expressed in nmol/L AMC released per min, or in mol/L substrate applied.

Fig.8 Hexactinellid silicatein sequence similarity to silicateins from demosponges and cathepsin (according to Müller et al. []). The silicatein protein from (SILCA_CRAME) was aligned with the silicatein-α from (SILCAa_SUBDO) and silicatein-β from (SILCAb_SUBDO), as well as with the cathepsin L sequence from hexactinellid (CATL_APHRVAS). Residues conserved (similar or related with respect to their physico-chemical properties) in all sequences are shown in white on black, and those in at least three sequences in black on gray. The characteristic sites in the sequences are marked; the catalytic triad (CT) amino acids, Ser in silicateins and Cys in cathepsin, and His and Asn. The borders within the mature silicatein (mature peptide) and the peptidase-C1 papain family cysteine protease domain (papain) are given. The “conventional” serine cluster (<<#Ser#>>), and the “-specific” serine cluster (<Ser>) are marked. The aa conserved in all freshwater sequences, here shown in the sequence, are listed above the alignment. A radial phylogenetic tree was constructed after the alignment of these five proteins, together with the two silicateins-α and-β (SILCAa_TETYA and SILCAb_TETYA), two isoforms (of silicatein-α) from (α-3) (SILCAa3_LUBAI and SILCAa1_LUBAI) as well as the three cathepsin sequences from (CAT1_CRAME; CAT2_CRAME and CAT3_CRAME) as well as the related papain-like cysteine peptidase XBCP3 from (PAPAIN_ARATH, AAK71314). The radial tree was constructed after the alignment. The numbers at the nodes are an indication of the level of confidence for the branches as determined by bootstrap analysis (1000 bootstrap replicates).

Fig.9 Model of silicateins. Based on the data available and an the algorithm of Ramachandran et al. [], the models have been predicted for the silicatein-α, silicatein-α3, and the silicatein from . The aa residues, comprising the catalytic triad (ct) Ser (aa22), His (aa161), and Asn (aa181) are highlighted in red. In addition, both the aa for the conserved Ser stretch (cSer) and the “hexactinellid-specific” Ser cluster (hSer) are in green. The “conventional” Ser cluster in all three sequences is found in the loop, flanking the alpha helix; the “hexactinellid-specific” Ser cluster exists in the outer loop of the exposed β-sheet.

Fig.10 Flexible breakage of a giant spicule (from ); the diameter of the fiber was 0.8 mm. The fibers were inserted into a traction device between two moving rubber wheels and were continuously pulled toward the fixation point (f) using a motor. The breakage was recorded with a high-speed camera. Flexible and stepwise breakage of a sponge spicule. The spicule had been completely bent by one circumvolution; the diameter of the circular wrapped around spicule was initially 4.5 cm. After proceeding traction, three stepwise breaking steps can be recorded ((B), (F), and (K)), which are interrupted by phases of flexible bending.

1	Kruse M, Müller I M, Muller W E G. Early evolution of metazoan serine/threonine and tyrosine kinases: Identification of selected kinases in marine sponges. Molecular Biology and Evolution , 1997, 14(12): 1326–1334
2	Kruse M, Leys S P, Müller I M, . Phylogenetic position of the hexactinellida within the phylum porifera based on the amino acid sequence of the protein kinase C from Rhabdocalyptus dawsoni. Journal of Molecular Evolution , 1998, 46(6): 721–728
3	Müller W E G, Wiens M, Adell T, , Bauplan of Urmetazoa: Basis for genetic complexity of Metazoa. In: International Review of Cytology – A Survey of Cell Biology, Vol 235 . San Diego: Elsevier Academic Press Inc, 2004, 53–92
4	Müller W E G, Li J H, Schr?der H C, . The unique skeleton of siliceous sponges (Porifera; Hexactinellida and Demospongiae) that evolved first from the Urmetazoa during the Proterozoic: a review. Biogeosciences , 2007, 4(2): 219–232
5	Pilcher H. Animal magnetism. Nature , 2005, 435(7045): 1022–1023
6	Murray J, Hjort J. The Depths of the Ocean. London: MacMillan, 1912
7	Schulze F E.Hexactinellida. Wissenschaftliche Ergebnisse der Deutschen Tiefsee-Expedition auf dem Dampfer “Valdivia” 1898–1899 . Stuttgart: Gustav Fischer Verlag, 1904
8	Roux M, Bouchet P, Bourseau J P, . L'environment bathyal au large de la Nouvelle-Calédonie: résultats preliminaries de la campagne CALSUB et consequences paléoécologiques. Geological Society of France , 1991, 162: 675–685
9	Müller W E G, Eckert C, Kropf K, . Formation of giant spicules in the deep-sea hexactinellid Monorhaphis chuni (Schulze 1904): electron-microscopic and biochemical studies. Cell and Tissue Research , 2007, 329(2): 363–378
10	Li J. Monorhaphis intermedia-a new species of Hexactinellida. Oceanologia et Limnologia Sinica , 1987, 18: 135–137
11	Tabachnick K R. Family Monorhaphididae Ijima, 1927. In: Hooper J N A, van Soest R. Systema Porifera: A Guide to the Classification of Sponges . New York: Kluwer Academic, 2002, 1264–1266
12	Wang X H, Li J H, Qiao L, . Structure and characteristics of giant spicules of the deep sea hexactinellid sponges of the genus Monorhaphis (Hexactinellida: Amphidiscosida: Monorhaphididae). Acta Zoologica Sinica , 2007, 53(3): 557–569
13	Sandford F. Physical and chemical analysis of the siliceous skeletons in six sponges of two groups (Demospongiae and Hexactinellida). Microscopy Research and Technique , 2003, 62(4): 336–355
14	Uriz M J, Turon X, Becerro M A, . Siliceous spicules and skeleton frameworks in sponges: Origin, diversity, ultrastructural patterns, and biological functions. Microscopy Research and Technique , 2003, 62(4): 279–299
15	Uriz M J. Mineral spiculogenesis in sponges. Canadian Journal of Zoology , 2006, 84: 322–356
16	Müller W E G, Jochum K P, Stoll B, . Formation of giant spicule from quartz glass by the deep sea sponge Monorhaphis. Chemistry of Materials , 2008, 20(14): 4703–4711
17	Müller W E G, Wang X H, Kropf K, . Bioorganic/inorganic hybrid composition of sponge spicules: Matrix of the giant spicules and of the comitalia of the deep sea hexactinellid Monorhaphis. Journal of Structural Biology , 2008, 161(2): 188–203
18	Levi C, Barton J L, Guillemet C, . A remarkably strong natural glassy rod — the anchoring spicule of the Monorhaphis sponge. Journal of Materials Science Letters , 1989, 8(3): 337–339
19	Müller W E G, Boreiko A, Schlossmacher U, . Identification of a silicatein(-related) protease in the giant spicules of the deep-sea hexactinellid Monorhaphis chuni. Journal of Experimental Biology , 2008, 211(3): 300–309
20	Müller W E G, Boreiko A, Wang X H, . Silicateins, the major biosilica forming enzymes present in demosponges: Protein analysis and phylogenetic relationship. Gene , 2007, 395(1–2): 62–71
21	Müller W E G, Rothenberger M, Boreiko A, . Formation of siliceous spicules in the marine demosponge Suberites domuncula. Cell and Tissue Research , 2005, 321(2): 285–297
23	Müller W E G, Schlossacher U, Wang X, . Poly(silicate)-metabolizing silicatein in siliceous spicules and silicasomes of demosponges comprises dual enzymatic activities (silica polymerase and silica esterase). FEBS Journal , 2008, 275(2): 362–370
24	Wang X H, Schloβmacher U, Jochum K P, . Silica-protein composite layers of the giant basal spicules from Monorhaphis: basis for their mechanical stability. Pure and Applied Chemistry , 2009 (in press)
25	Sumerel J L, Morse D E. Biotechnological advances in biosilicification. In: Silicon Biomineralization, Vol 33 . Berlin: Springer-Verlag Berlin, 225–247
26	Shimizu K, Cha J, Stucky G D, . Silicatein alpha: Cathepsin L-like protein in sponge biosilica. Proceedings of the National Academy of Sciences of the United States of America , 1998, 95(11): 6234–6238
27	Cha J N, Shimizu K, Zhou Y, . Silicatein filaments and subunits from a marine sponge direct the polymerization of silica and silicones in vitro. Proceedings of the National Academy of Sciences of the United States of America , 1999, 96(2): 361–365
28	Krasko A, Lorenz B, Batel R, . Expression of silicatein and collagen genes in the marine sponge Suberites domuncula is controlled by silicate and myotrophin. European Journal of Biochemistry , 2000, 267(15): 4878–4887
29	Müller W E G, Wang X H, Kropf K, . Silicatein expression in the hexactinellid Crateromorpha meyeri: the lead marker gene restricted to siliceous sponges. Cell and Tissue Research , 2008, 333(2): 339–351
30	Müller W E, Krasko A, Le Pennec G, . Molecular mechanism of spicule formation in the demosponge Suberites domuncula: silicatein-collagen-myotrophin. Progress in Molecular and Subcellular Biology , 2003, 33: 195–221
31	Wiens M, Belikov S I, Kaluzhnaya O V, . Molecular control of serial module formation along the apical-basal axis in the sponge Lubomirskia baicalensis: silicateins, mannose-binding lectin and mago nashi. Development Genes and Evolution , 2006, 216(5): 229–242
32	Müller W E G, Boreiko A, Schlossmacher U, . Fractal-related assembly of the axial filament in the demosponge Suberites domuncula: Relevance to biomineralization and the formation of biogenic silica. Biomaterials , 2007, 28(30): 4501–4511
33	Ramachandran G N, Ramakrishnan C, Sasisekharan V. Stereochemistry of polypeptide chain configurations. Journal of Molecular Biology , 1963, 7(1): 95–99
34	Robinson P N. A Java program for drawing Ramachandran plots. peter.robinson@charite.de, 2007
35	Mayer G. Rigid biological systems as models for synthetic composites. Science , 2005, 310(5751): 1144–1147
36	Mayer G, Trejo R, Lara-Curzio E, . Lessons for new classes of inorganic/organic composites from the spicules and skeleton of the sea sponge Euplectella aspergillum. Mechanical Properties of Bioinspired and Biological Materials , 2005, 844: 79–86
37	Perovic S, Krasko A, Prokic I, . Origin of neuronal-like receptors in Metazoa: cloning of a metabotropic glutamate GABA-like receptor from the marine sponge Geodia cydonium. Cell and Tissue Research , 1999, 296(2): 395–404
38	Chevreux B, Pfisterer T, Drescher B, . Using the miraEST assembler for reliable and automated mRNA transcript assembly and SNP detection in sequenced ESTs. Genome Research , 2004, 14(6): 1147–1159
39	Pavans de Ceccatty M. Coordination in sponges — foundations of integration. American Zoologist , 1974, 14(3): 895–903
40	Mackie G O. Is there a conduction system in sponges? Colloq Int Centre Natl Res Sci , 1979, 291: 145–151
41	Leys S P, Degnan B M. Cytological basis of photoresponsive behavior in a sponge larva. Biological Bulletin , 2001, 201(3): 323–338
42	Leys S P, Cronin T W, Degnan B M, . Spectral sensitivity in a sponge larva. Journal of Comparative Physiology A — Neuroethology Sensory Neural and Behavioral Physiology , 2002, 188(3): 199–202
43	Cattaneo-Vietti R, Bavestrello G, Cerrano C, . Optical fibres in an Antarctic sponge. Nature , 1996, 383(6599): 397–398
44	Aizenberg J, Sundar V C, Yablon A D, . Biological glass fibers: Correlation between optical and structural properties. Proceedings of the National Academy of Sciences of the United States of America , 2004, 101(10): 3358–3363
45	Müller W E G, Wendt K, Geppert C, . Novel photoreception system in sponges? Unique transmission properties of the stalk spicules from the hexactinellid Hyalonema sieboldi. Biosensors & Bioelectronics , 2006, 21(7): 1149–1155
46	Murr M M, Morse D E. Fractal intermediates in the self-assembly of silicatein filaments. Proceedings of the National Academy of Sciences of the United States of America , 2005, 102(33): 11657–11662
47	Krasko A, Schr?der H C, Batel R, . Iron induces proliferation and morphogenesis in primmorphs from the marine sponge Suberites domuncula. DNA and Cell Biology , 2002, 21(1): 67–80
48	Schr?der H C, Perovic-Ottstadt S, Wiens M, . Differentiation capacity of epithelial cells in the sponge Suberites domuncula. Cell and Tissue Research , 2004, 316(2): 271–280

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