Hostname: page-component-cd9895bd7-dzt6s Total loading time: 0 Render date: 2024-12-29T12:12:36.851Z Has data issue: false hasContentIssue false

Preliminary characterization of the lipid and protein components of the protective surface membranes of a pentastomid Porocephalus crotali

Published online by Cambridge University Press:  06 April 2009

D. A. C. Jones
Affiliation:
Department of Biological Sciences, The University of Dundee, Dundee DD1 4HN, Scotland
R. J. Henderson
Affiliation:
NERC Unit of Aquatic Biochemistry, Department of Biological Sciences, University of Stirling, Stirling FK9 4LA, Scotland
J. Riley
Affiliation:
Department of Biological Sciences, The University of Dundee, Dundee DD1 4HN, Scotland

Extract

All instars of the pentastomid Porocephalus crotali, in the tissues of rat intermediate hosts and the lung of rattlesnake definitive hosts, are covered by a vesicular or stacked membranous secretory product which is synthesized in sub-parietal cells (SPC) and channelled to the cuticle via multitudinous chitin-lined ducts. In rats this enveloping foam of vesicles survives for the duration of a cuticle. We have purified and partially characterized the lipid and protein composition of lamellate droplets from SPC of infective nymphs. Lipids in the droplets comprised a mixture of neutral and polar lipid classes with cholesterol being the major neutral lipid and phosphatidylcholine the dominant polar lipid. In addition, Triton X-114 phase separation of the protein component of droplets partitioned these into an aqueous phase (with a major band at 60 kDa), a detergent phase with two hydrophobic polypeptides (24 and 16 kDa) and an insoluble pellet containing several minor proteins and major bands at 31 and 107 kDa. Western blotting, with rabbit anti-lamellate droplet antiserum, strongly label only the 60 kDa and the two hydrophobic proteins. Significantly no proteins at all label with serum from infected rats from 50–150 days post-infection, a finding endorsed by indirect fluorescent antibody tests on sectioned V–VII stage nymphs. Possible immunomodulatory functions of SPC-derived surface membranes are discussed and, in this regard, they are compared with pulmonary surfactant, a product of alveolar Type II cells which lines all vertebrate lungs.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1992

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Abele, L. G., Kim, W. & Felgenhauer, B. E. (1989). Molecular evidence for the inclusion of the phylum Pentastomida in the Crustacea. Molecular and Biochemical Evolution 6, 685–91.Google Scholar
Ambrose, N. C. & Riley, J. (1988 a). Light microscope observations of granulomatous reactions against developing Porocephalus crotali (Pentastomida: Porocephalida) in mouse and rat. Parasitology 97, 115.Google Scholar
Ambrose, N. C. & Riley, J. (1988 b). Fine structural aspects of secretory processes in a pentastomid arthropod parasite in its mouse and rattlesnake hosts. Tissue and Cell 20, 381404.CrossRefGoogle Scholar
Ambrose, N. C. & Riley, J. (1988 c). Studies on the host parasite interface during the development of a pentastomid arthropod parasite in rodent intermediate hosts, with observations on protective surface membranes. Tissue and Cell 20, 721–44.CrossRefGoogle ScholarPubMed
Ambrose, N. C. & Riley, J. (1989). Further evidence for the protective role of sub-parietal cell membranous secretory product on the cuticle of a pentastomid arthropod parasite developing in its rodent intermediate host. Tissue and Cell 21, 699722.Google Scholar
Ansfield, M. J., Kaltreider, H. B., Benson, B. J. & Coldwell, J. L. (1979). Immunosuppressive activity of canine surface active materials. Journal of Immunology 122, 1062–6.Google Scholar
Baer, J. G. (1952). Ecology of Animal Parasites. Urbana: University of Illinois Press.Google Scholar
Bordier, C. (1981). Phase separation of integral membrane proteins in Triton X-114 solution. Journal of Biological Chemistry 230, 1604–7.Google Scholar
Christie, W. W. (1982). Lipid Analysis. Oxford: Pergamon Press.Google Scholar
Esslinger, J. H. (1962). Development of Porocephalus crotali (Humboldt, 1808) (Pentastomida) in experimental intermediate hosts. Journal of Parasitology 48, 452–6.Google Scholar
Goerke, J. (1974). Lung surfactant. Biochimica et Biophysica Acta 334, 241–61.CrossRefGoogle Scholar
Heukeshoven, J. & Dernick, R. (1988). Improved silver staining procedure for fast staining in PhastSystem development Unit I. Staining of sodium dodecyl sulfate gels. Electrophoresis 9, 60–1.CrossRefGoogle ScholarPubMed
Hill, B. (1988). Biology of Surfactants. Cambridge: Cambridge University Press.Google Scholar
Jones, D. A. C., Riley, J., Kerby, N. W. & Knox, D. P. (1991). Isolation and preliminary characterization of a 48-kilodalton metalloproteinase from the excretory/secretory components of the frontal glands of Porocephalus pentastomids. Molecular and Biochemical Parasitology 46, 6172.Google Scholar
Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of the bacteriophage T4. Nature, London 227, 680–5.Google Scholar
Oosterlaken-Dijksterhuis, M. A., Eijk, M. Van, Buel, B. L. M. Van, Golde, L. M. G. Van & Haagsman, H. P. (1991). Surfactant protein composition of lamellar bodies isolated from rat lung. The Biochemical Journal 274, 115–19.Google Scholar
Rich, E. A. (1990). Pulmonary surfactant as a physiologic suppressive agent. Journal of Laboratory and Clinical Medicine 116, 45.Google Scholar
Richman, P. S., Batcher, S. & Catanzaro, A. (1990). Pulmonary surfactant suppresses the immune lung injury response to inhaled antigen in guinea pigs. Journal of Laboratory and Clinical Medicine 116, 1826.Google ScholarPubMed
Riley, J. (1981). Some observations on the development of Porocephalus crotali (Pentastomida: Porocephalida) in the Western Diamondback Rattlesnake (Crotalus atrox). International Journal for Parasitology 11, 127–31.Google Scholar
Riley, J., James, J. L. & Banaja, A. A. (1979). The possible role of the frontal and sub-parietal gland systems of the pentastomid Reighardia sternae (Diesing, 1864) in the evasion of the host immune response. Parasitology 78, 5366.CrossRefGoogle ScholarPubMed
Sitrin, R. G., Ansfield, M. J. & Kaltreider, H. B. (1985). The effect of pulmonary surface active material on the generation of murine B and T lymphocytes in vitro. Experimental Lung Research 9, 8597.CrossRefGoogle Scholar
Towbin, H., Staehelin, T. & Gordon, J. (1979). Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some application. Proceedings of the National Academy of Sciences, USA 76, 313–40.CrossRefGoogle Scholar
Wilsher, M. L., Hughes, D. A. & Haslam, P. L. (1988). Immunoregulatory properties of pulmonary surfactant: effect of lung lining fluid on proliferation of human lymphocytes. Thorax 43, 354–9.Google Scholar
Wilsher, M. L., Parker, D. J. & Haslam, P. L. (1990). Immunosuppression by pulmonary surfactant: mechanisms of action. Thorax 45, 38.CrossRefGoogle ScholarPubMed