Ultrastructural characterization of antennal sensilla and immunocytochemical localization of a chemosensory protein in Carausius morosus Brünner (Phasmida: Phasmatidae).

The aim of this work was to investigate the olfactory system of the walking stick insect, Carausius morosus. Morphological, ultrastructural and immunocytochemical studies of adult female antennae were conducted by scanning and transmission electron microscopy. Extensive cross-section series were made through the last antennal segment to define the cuticular apparatus, wall pore distribution and the number of innervating receptor neurons of each sensillum type. Single-walled wall pore sensilla occur in three subtypes: (i) with 27 or 28 branched receptor neurons, (ii) with two branched neurons and (iii) with one or two unbranched neurons, respectively. Double-walled wall pore sensilla were found in two subtypes with spoke channels, one with four unbranched neurons, the other with two unbranched neurons. One terminal pore sensillum was found, showing two cavities within the hair and being innervated by six sensory cells. Immunocytochemical experiments were performed to show the localization of a 19 kDa soluble protein found in the chemosensory organs of C. morosus. This protein shows an amino acid sequence homologous to the family of chemosensory proteins (CSP). The polyclonal antibody raised against the purified protein (CSP-cmA) showed, for the first time in CSPs, a strong labeling in olfactory sensilla, specifically in the sensillum lymph surrounding the dendritic branches of SW-WP sensilla and in the uninnervated lumen between the two concentric walls of DW-WP type 1 sensilla.

[1]  J. Nardi,et al.  Diversity of odourant binding proteins revealed by an expressed sequence tag project on male Manduca sexta moth antennae , 1999, Insect molecular biology.

[2]  C. Masson,et al.  Cloning and Expression of a Queen Pheromone-Binding Protein in the Honeybee: an Olfactory-Specific, Developmentally Regulated Protein , 1999, The Journal of Neuroscience.

[3]  R. Steinbrecht,et al.  Mechanosensitive and Olfactory Sensilla of Insects , 1984 .

[4]  B. Hansson,et al.  Fine structure and distribution of antennal sensilla of the desert locust, Schistocerca gregaria (Orthoptera: Acrididae) , 1998, Cell and Tissue Research.

[5]  R. Steinbrecht Freeze‐Substitution for morphological and immunocytochemical studies in insects , 1993, Microscopy research and technique.

[6]  A. Scaloni,et al.  Purification, structural characterization, cloning and immunocytochemical localization of chemoreception proteins from Schistocerca gregaria. , 1999, European journal of biochemistry.

[7]  R. Steinbrecht Pore structures in insect olfactory sensilla: A review of data and concepts , 1997 .

[8]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[9]  J. Picimbon,et al.  Olfactory soluble proteins of cockroaches , 1999 .

[10]  T. Close,et al.  Isolation of the origin of replication of the IncW-group plasmid pSa. , 1982, Gene.

[11]  G. Danscher Localization of gold in biological tissue , 1981, Histochemistry.

[12]  H. Sass,et al.  Relationship between structure and function of antennal chemo-, hygro-, and thermoreceptive sensilla in Periplaneta americana , 1977, Cell and Tissue Research.

[13]  H. Tichy,et al.  Lamellated outer dendritic segments of a sensory cell within a poreless thermo- and hygroreceptive sensillum of the insect Carausius morosus , 1978, Cell and Tissue Research.

[14]  R. Vogt,et al.  Odorant binding protein diversity and distribution among the insect orders, as indicated by LAP, an OBP-related protein of the true bug Lygus lineolaris (Hemiptera, Heteroptera). , 1999, Chemical senses.

[15]  G. Hasan,et al.  Expression patterns of two putative odorant-binding proteins in the olfactory organs of Drosophila melanogaster have different implications for their functions , 2000, Cell and Tissue Research.

[16]  E H Slifer Sense organs on the antennal flagellum of a walkingstick Carausius morosus Brünner (Phasmida) , 1966, Journal of morphology.

[17]  C. Masson,et al.  Identification and developmental profiles of hexamerins in antenna and hemolymph of the honeybee, Apis mellifera. , 1998, Insect biochemistry and molecular biology.

[18]  P. Lucas,et al.  Functional characterization of a new class of odorant-binding proteins in the moth Mamestra brassicae. , 1998, Biochemical and biophysical research communications.

[19]  P. Pelosi,et al.  Odorant-binding proteins in insects. , 1995, Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology.

[20]  A. Scaloni,et al.  Purification and molecular cloning of chemosensory proteins from Bombyx mori. , 2000, Archives of insect biochemistry and physiology.

[21]  G. Ziegelberger,et al.  Immunolocalization of pheromone-binding protein and general odorant-binding protein in olfactory sensilla of the silk moths Antheraea and Bombyx , 1995, Cell and Tissue Research.

[22]  M. Laue Immunolocalization of general odorant-binding protein in antennal sensilla of moth caterpillars. , 2000, Arthropod structure & development.

[23]  R. Steinbrecht,et al.  Chemosensory proteins from the proboscis of mamestra brassicae. , 2000, Chemical senses.

[24]  R. Steinbrecht,et al.  Fine structure of the antennal receptors of the bed bug, Cimex lectularius L. , 1976, Tissue & cell.

[25]  J. Carlson,et al.  Olfactory Coding in a Compound Nose: Coexpression of Odorant‐Binding Proteins in Drosophila a , 1998, Annals of the New York Academy of Sciences.

[26]  M. McKenna,et al.  Putative Drosophila pheromone-binding proteins expressed in a subregion of the olfactory system. , 1994, The Journal of biological chemistry.

[27]  E. Eguchi,et al.  Atlas of arthropod sensory receptors : dynamic morphology in relation to function , 1999 .

[28]  H. Altner,et al.  Ultrastructure of invertebrate chemo-, thermo-, and hygroreceptors and its functional significance. , 1980 .

[29]  F. Bin,et al.  Antennal chemosensilla and glands, and their possible meaning in the reproductive behaviour of Trissolcus basalis (Woll.) (Hym. : Scelionidae) , 1989 .

[30]  Franz Ameismeier Ultrastructure of the chemosensitive basiconic single-walled wall-pore sensilla on the antennae in adults and embryonic stages of Locusta migratoria L. (Insecta, Orthoptera) , 1987, Cell and Tissue Research.

[31]  G. Stange,et al.  Molecular cloning, by a novel approach, of a cDNA encoding a putative olfactory protein in the labial palps of the moth Cactoblastis cactorum. , 1997, Gene.

[32]  R. Steinbrecht,et al.  Functional morphology of a double-walled multiporous olfactory sensillum: the sensillum coeloconicum of Bombyx mori (Insecta, Lepidoptera). , 1998, Tissue & cell.

[33]  R. Vogt,et al.  Intergeneric distribution and immunolocalization of a putative odorant-binding protein in true bugs (Hemiptera, Heteroptera). , 1998, The Journal of experimental biology.

[34]  R. Steinbrecht Functional Morphology of Pheromone-Sensitive Sensilla , 1987 .

[35]  A. Scaloni,et al.  Soluble proteins from chemosensory organs of Eurycantha calcarata (Insects, Phasmatodea). , 2000, Insect biochemistry and molecular biology.

[36]  R. Steinbrecht,et al.  Volume and surface of receptor and auxiliary cells in hygro-/thermoreceptive sensilla of moths (Bombyx mori, Antheraea pernyi, and A. polyphemus) , 2004, Cell and Tissue Research.

[37]  P. Pelosi,et al.  Soluble proteins in chemosensory organs of phasmids. , 1996, Insect biochemistry and molecular biology.

[38]  H. Breer,et al.  Chemosensory proteins of Locusta migratoria (Orthoptera: Acrididae). , 2000, Insect biochemistry and molecular biology.

[39]  R. Chapman The Insects: Structure and Function , 1969 .

[40]  R. Steinbrecht,et al.  Odorant‐Binding Proteins: Expression and Function , 1998, Annals of the New York Academy of Sciences.

[41]  M. Rosbash,et al.  Members of a family of drosophila putative odorant-binding proteins are expressed in different subsets of olfactory hairs , 1994, Neuron.

[42]  M. Karnovsky,et al.  A formaldehyde-glutaraldehyde fixative of high osmolality for use in electron-microscopy , 1965 .

[43]  P. Pelosi,et al.  Putative odorant-binding protein in antennae and legs of Carausius morosus (Insecta, Phasmatodea). , 1996, Insect biochemistry and molecular biology.