Hostname: page-component-586b7cd67f-2plfb Total loading time: 0 Render date: 2024-11-30T15:33:10.989Z Has data issue: false hasContentIssue false
  • English
  • Français

Histochemical studies on microfilariae

Published online by Cambridge University Press:  06 April 2009

M. G. Simpson  and
B. R. Laurence
M. G. Simpson
Affiliation:
Department of Entomology, London School of Hygiene and Tropical Medicine
B. R. Laurence
Affiliation:
Department of Entomology, London School of Hygiene and Tropical Medicine
Get access Rights & Permissions [Opens in a new window]

Extract

Microfilariae belonging to the genera Brugia, Loa and Wuchereria have been stained by a wide variety of histochemical methods. The results are recorded in eight Tables.

The sheath of the microfilaria contains polyanionic material, probably polysulphate, periodate reactive mucosubstance, sulphur-protein and complex lipids. The fluid within the sheath is also polyanionic.

The cuticle of the microfilaria is stabilized by disulphide bonds possibly in a collagen but it was not possible to demonstrate collagen by staining. The cuticle also contains a periodate reactive G mucosubstance and complex lipids. The annular grooves of the cuticle also contain a periodate unreactive mucosubstance. Certain areas of the cuticle, around the excretory pore, the pores of the Schwanzgebilde, and the cephalic structures - oral ring, hook and spines - are hardened by deposition of a keratinous scleroprotein. Some reactions of the cuticle may be masked by the presence of basic protein.

The excretory vesicle contains polyanionic material but there was no clear evidence that the polyanionic material within the fluid in the sheath came from the excretory vesicle. The excretory vesicle, the anal vesicle and the subcuticular cells contain periodate reactive G mucosubstance. Most RNA staining was found in the excretory vesicle, the excretory cell, the anal vesicle, the rectal cells and the G cell.

The pharyngeal thread, the Innenkörper, the rectal cells and the anal vesicle represent the modified gut of this first stage larva. The pharyngeal thread appears to consist largely of a protein-mucosubstance complex that can be unmasked by oxidation. The Innenkörper contains a basic protein and acidic mucosubstance, possibly sialomucin, complex that can be unmasked by adjusting the electrolyte concentration of the staining media.

The relevance of some of these techniques in the analysis of possibly unique invertebrate structures is discussed briefly.

We thank Dr D. A. Denham, Mr P. R. N. Pester and Dr T. Ponnudurai in particular, and many other correspondents, for supplying us with microfilariae for staining. This work was supported by a grant from the Medical Research Council of Great Britain.

Type
Research Article
Information
Parasitology , Volume 64 , Issue 1 , February 1972 , pp. 61 - 88
Copyright
Copyright © Cambridge University Press 1972

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

REFERENCES

Abe, Y., Hori, E., Yamaji, T., Ono, T., Ushuku, F. & Kagei, N. (1960). Studies on filaria and filariasis. Morphological and histochemical studies on microfilaria of Wuchereria bancrofti. Medical Journal of Kagoshima University 12, 1098–114.Google Scholar
Adams, C. W. M. & Bayliss, O. B. (1968). Histochemistry of myelin. VI. Solvent action of acetone on brain and other lipid rich tissues. Journal of Histochemistry and Cytochemistry 16, 115–18.CrossRefGoogle ScholarPubMed
Barka, T. & Anderson, P. J. (1965). Histochemistry, Theory, Practice and Bibliography. New York, Evanston and London: Harper and Row.Google Scholar
Bloom, G. & Kelly, J. W. (1960). The copper phthalocyanine dye ‘Astrablau’ and its staining properties, especially the staining of mast cells. Histochemie 2, 48–57.CrossRefGoogle ScholarPubMed
Buckley, J. J. C. (1955). Symposium on loiasis. V. The morphology of the larval stages in the vector: some of the problems involved. Transactions of the Royal Society of Tropical Medicine and Hygiene 49, 122–6.Google Scholar
Bussolati, G., Rost, F. W. D. & Pearse, A. G. E. (1969). Fluorescence metachromasia in polypeptide hormone producing cells of the APUD series, and its significance in relation to the structure of the precursor protein. Histochemical Journal 1, 517–30.CrossRefGoogle Scholar
Cameron, M. L. & Steele, J. E. (1959). Simplified aldehyde fuchsin staining of neurosecretory cells. Stain Technology 34, 265–6.CrossRefGoogle ScholarPubMed
Casselman, W. B. (1962). Histochemical Technique. London: Methuen.Google Scholar
Chowdhury, A. B., Dasgupta, B., Ray, H. N. & Bhaduri, N. V. (1956). Histochemical pattern of the microfilaria of Wuchereria bancrofti in peripheral blood. Bulletin of the Calcutta School of Tropical Medicine 4, 23–4.Google Scholar
Curren, R. C. (1964). The histochemistry of mucopolysaccharides. International Review of Cytology 17, 149–212.CrossRefGoogle Scholar
Elder, H. Y. & Owen, G. (1967). Occurrence of elastic fibres in invertebrates. Journal of Zoology 152, 1–8.CrossRefGoogle Scholar
Feng, Lan-chou (1933). A comparative study of the anatomy of Microfilaria malayi Brug, 1927 and Microfilaria bancrofti Cobbold, 1877. Chinese Medical Journal 47, 1214–46.Google Scholar
Frank, W. (1964). Die Entwicklung von Macdonaldius oschei Chabaud et Frank 1961 (Filarioidea, Onchocercidae) in der Lederzecke Ornithodoros talaje Guerin-Meneville (Ixodoidea, Argasidae). Zeitshrift fur Parasitenkunde 24, 319–50.Google Scholar
French, J. E. & Benditt, E. P. (1953). The histochemistry of connective tissue. II. The effect of proteins on the selective staining of mucopolysaccharides by basic dyes. Journal of Histochemistry and Cytochemistry 1, 321–5.CrossRefGoogle ScholarPubMed
Fujimoto, D. (1968). Correlation between pronase susceptibility and hydroxy-proline content of collagen fibre. Biochimica et Biophysica Acta 154, 183–9.CrossRefGoogle Scholar
Fülleborn, F. (1929). Filariosen des Menschen. In Handbuch der pathogenen Mikroorganismen 6, 1039–224.Google Scholar
Fullmer, H. M. (1960). Effect of peracetic acid on the enzymatic digestion of various mucopolysaccharides: reversal of the PAS staining reaction of mucin. Journal of Histochemistry and Cytochemistry 8, 113–21.CrossRefGoogle ScholarPubMed
Fullmer, H. M. & Lillie, R. D. (1957). The staining of collagen with elastic tissue stains. Journal of Histochemistry and Cytochemistry 5, 11–14.CrossRefGoogle ScholarPubMed
Fullmer, H. M. & Lillie, R. D. (1958). The oxytalan fibre: a previously undescribed connective tissue fibre. Journal of Histochemistry and Cytochemistry 6, 425–30.CrossRefGoogle Scholar
Glick, D. (1967). Usage of ‘histochemical’, ‘staining’ and ‘biochemical’ in histochemical literature. Journal of Histochemistry and Cytochemistry 15, 299.CrossRefGoogle ScholarPubMed
Green, J. A. (1960). Digestion of collagen and reticulin in paraffin section by collagenase. Stain Technology 35, 273–6.CrossRefGoogle ScholarPubMed
Gurr, E. (1958). Methods of Analytical Histology and Histochemistry. London: Leonard Hill.Google Scholar
Hadler, W. A., Ziti, L. M., Patelli, A. S., Vozza, J. A. & De Lucca, O. (1968). Histochemical meaning of the chrome-alum hematoxylin and the aldehyde fuchsin staining: An investigation accomplished by a spot test technique carried on filter paper models. Acta Histochemica 29, 320–38.Google ScholarPubMed
Hadler, W. A., Ziti, L. M., De Lucca, O. & Patelli, A. S. (1968). A new technique for histochemical detection of cholesterol-like compounds. Acta Histochemica 30, 70–8.Google ScholarPubMed
Hale, A. J. (1957). The histochemistry of polysaccharides. International Review of Cytology 6, 193–263.CrossRefGoogle Scholar
Hockley, D. J., Worms, M. J. & Ebden, L. M. (1968). Electron micrographs of microfilariae. Transactions of the Royal Society of Tropical Medicine and Hygiene 62, 468–9.Google Scholar
Hunt, S. (1970). Polysaccharide-Protein Complexes in Invertebrates. London and New York: Academic Press.Google Scholar
Johnson, K. H. & Bemrick, W. J. (1969). Dirofilaria immitis microfilariae: electron microscopic examination of cuticle and muscle cells. American Journal of Veterinary Research 30, 1443–50.Google ScholarPubMed
Kvistberg, D. R. (1966). Infra-red spectrophotometry. Its use in characterising histochemical reaction mechanisms. Journal of Histochemistry and Cytochemistry 14, 853–61.CrossRefGoogle Scholar
Landing, B. H., Hall, H. E. & West, C. D. (1956). Aldehyde-fuchsin-positive material of the posterior pituitary. Laboratory Investigation 5, 256–66.Google ScholarPubMed
Laurence, B. R. & Pester, F. R. N. (1967). Adaptation of a filarial worm, Brugia patei, to a new mosquito host, Aedes togoi. Journal of Helminthology 41, 365–92.CrossRefGoogle Scholar
Laurence, B. R. & Simpson, M. G. (1968). Cephalic and pharyngeal structures in microfilariae revealed by staining. Journal of Helminthology 42, 309–30.CrossRefGoogle ScholarPubMed
Laurence, B. R. & Simpson, M. G. (1969). Staining microfilariae for structures other than nuclei. Transactions of the Royal Society of Tropical Medicine and Hygiene 63, 801–10.CrossRefGoogle ScholarPubMed
Laurence, B. R. & Simpson, M. G. (1971). The microfilaria of Brugia: a first stage nematode larva. Journal of Helminthology 45, 23–40.CrossRefGoogle ScholarPubMed
Leppi, T. J. & Stoward, P. J. (1965). On the use of testicular hyaluronidase for identifying acid mucins in tissue sections. Journal of Histochemistry and Cytochemistry 13, 406–7.CrossRefGoogle ScholarPubMed
Lev, R. & Spicer, S. S. (1964). Specific staining of sulphate groups with alcian blue at low pH. Journal of Histochemistry and Cytochemistry 12, 309.CrossRefGoogle ScholarPubMed
Lhotka, J. F. (1952). Histochemical use of sodium bismuthate. Stain Technology 27, 259–62.CrossRefGoogle ScholarPubMed
Lhotka, J. F. (1957). Histochemical use of lead tetra acetate. II. α-hydroxy-carboxylic acid localisation. Stain Technology 32, 275–8.CrossRefGoogle Scholar
Lillie, R. D. (1965). Histopathologic Technic and Practical Histochemistry. New York: McGraw-Hill.Google Scholar
Lillie, R. D., Bangle, R. & Fisher, E. R. (1954). Metachromatic basophilia of keratin after oxidation-cleavage of disulphide bonds. Journal of Histochemistry and Cytochemistry 2, 95–102.CrossRefGoogle Scholar
Lumsden, R. D., Oaks, J. A. & Alworth, W. L. (1970). Cytological studies on the absorptive surfaces of cestodes. IV. Localisation and cytochemical properties of membrane-fixed cation binding sites. Journal of Parasitology 56, 736–47.CrossRefGoogle ScholarPubMed
Lyons, K. M. (1970). The fine structure and function of the adult epidermis of two skin parasitic monogeneans, Entobdella soleae and Ancanthocotyle elegans. Parasitology 60, 39–52.CrossRefGoogle ScholarPubMed
Mancini, R. E., Vilar, O., Stein, E. & Fiorini, H. (1961). A histochemical and radio-autographic study of the participation of fibroblasts in the production of mucopolysaccharides in connective tissue. Journal of Histochemistry and Cytochemistry 9, 278–91.CrossRefGoogle Scholar
Mander, S. T. K., Mander, L. N. & Carmichael, G. G. (1968). The staining mechanism of aldehyde-fuchsin, with reference to the oxtyalan fiber in the mouse. Journal of Histochemistry and Cytochemistry 16, 480–5.CrossRefGoogle Scholar
Materazzi, G. & Ferretti, E. (1970). Histochemical and histophysical investigations on the acetylation blockade of carboxylic groups of polysaccharides. Journal of Histochemistry and Cytochemistry 18, 504–9.CrossRefGoogle ScholarPubMed
Materazzi, G. & Vitaioli, L. (1969). Further cytochemical studies of the mucopolysaccharides of the test cells of Ciona intestinalis L. Histochemie 19, 58–63.CrossRefGoogle ScholarPubMed
Mathews, M. B. (1967). Macromolecular evolution of connective tissue. Biological Reviews 42, 499–551.CrossRefGoogle ScholarPubMed
McBride, O. W. & Harrington, W. F. (1965). Evidence for disulphide cross linkages in an invertebrate collagen. Journal of Biological Chemistry 240, 4545–7.CrossRefGoogle Scholar
McManus, J. F. A. & Mowry, R. W. (1960). Staining Methods, Histologic and Histochemical. New York, Evanston and London: Harper and Row.Google Scholar
Meyer, K. (1966). Problems of histochemical identification of carbohydrate rich tissue components. Journal of Histochemistry and Cytochemistry 14, 605–6.CrossRefGoogle ScholarPubMed
Mowry, R. W. (1963). The special value of methods that colour both acidic and vic hydroxyl groups in the histochemical study of mucins with revised directions for the colloidal iron stain, the use of alcian blue 8GX and their combinations with the PAS reaction. Annals of the New York Academy of Science 106, 402–23.CrossRefGoogle Scholar
Neurath, H. (1966). The Proteins. Vol. 4. Composition, Structure and Function. New York and London: Academic Press.Google Scholar
Paget, G. E. & Eccleston, E. (1959). Aldehyde thionin. A stain having similar properties to aldehyde fuchsin. Stain Technology 34, 223–6.CrossRefGoogle Scholar
Pautard, F. G. E. & Zola, H. (1967). The location of onuphic acid in Hyalinoecia tubicola. Journal of Histochemistry and Cytochemistry 15, 737–44.CrossRefGoogle ScholarPubMed
Peace, D. C. (1966). Polysaccharides associated with the exterior surface of epithelial cells, kidney, intestine, brain. Journal of Ultrastructure Research 15, 555–88.CrossRefGoogle Scholar
Pearse, A. G. E. (1968). Histochemistry, Theoretical and Applied. London: Churchill.Google Scholar
Quintarelli, G. (1963). Masking action of basic proteins on sialic acid carboxyls in epithelial mucins. Experimentia 19, 230–1.CrossRefGoogle ScholarPubMed
Quintarelli, G. (1968). The Chemical Physiology of Mucopolysaccharides. Boston: Little, Brown and Co.Google Scholar
Rees, D. A. (1970). No bones about polymers. Nature, London 227, 1274.CrossRefGoogle Scholar
Ringertz, N. & Zetterberg, A. (1966). Cytochemical demonstration of histones and protamines. Mechanism and specificity of the alkaline bromphenol blue binding reaction. Experimental Cell Research 42, 260–4.CrossRefGoogle ScholarPubMed
Saunders, A. M. & Rosan, R. C. (1966). Histochemical nomenclature of carbohydrate-rich components. Journal of Histochemistry and Cytochemistry 14, 869.CrossRefGoogle ScholarPubMed
Scott, J. E. & Dorling, J. (1965). Differential staining of acid glycosaminoglycans (mucopolysaccharides) by alcian blue in salt solutions. Histochemie 5, 221–33.CrossRefGoogle ScholarPubMed
Scott, J. E., Dorling, J. & Stockwell, R. A. (1968). Reversal of protein blocking of basophilia in salt solutions: implications in the localisation of polyanions using alcian blue. Journal of Histochemistry and Cytochemistry 16, 383–6.CrossRefGoogle ScholarPubMed
Scott, J. E. & Willett, I. H. (1966). Binding of cationic dyes to nucleic acids and other biological polyanions. Nature, London 209, 985.CrossRefGoogle Scholar
Silverman, L. & Glick, D. (1966). Studies in histochemistry. LXXXV. Histochemical demonstration of protein with bromsulfalein. Journal of Histochemistry and Cytochemistry 14, 425–6.CrossRefGoogle ScholarPubMed
Solcia, E., Vasallo, G. & Capella, C. (1968). Selective staining of endocrine cells by basic dyes after acid hydrolysis. Stain Technology 43, 257–63.CrossRefGoogle ScholarPubMed
Spicer, S. S. (1961). The use of various cationic agents in histochemical differentiation of mucopolysaccharides. American Journal of Clinical Pathology 36, 393–407.CrossRefGoogle Scholar
Spicer, S. S. (1965). Diamine methods for differentiating mucosubstances histochemically. Journal of Histochemistry and Cytochemistry 13, 211–34.CrossRefGoogle Scholar
Spicer, S. S. & Henson, J. G. (1967). Methods for localising mucosubstances in epithelial and connective tissues. In Methods and Achievements in Experimental Pathology, ed. Bajurz, E. and Jasmin, G..Google Scholar
Spicer, S. S., Horn, R. G. & Leppi, T. J. (1967). Histochemistry of connective tissue mucopolysaccharides. In The Connective Tissue. International Academy of Pathology Monograph No. 7. Baltimore.Google Scholar
Spicer, S. S. & Jarrels, M. H. (1961). Histochemical reaction of an aromatic diamine with acid groups and periodate engendered aldehydes in mucopolysaccharides. Journal of Histochemistry and Cytochemistry 9, 368–79.CrossRefGoogle ScholarPubMed
Spicer, S. S., Leppi, T. J. & Stoward, P. J. (1965). Suggestions for a histochemical terminology of carbohydrate rich components. Journal of Histochemistry and Cytochemistry 13, 599–603.CrossRefGoogle Scholar
Spicer, S. S. & Meyer, D. B. (1960). Histochemical differentiation of acid mucopolysaccharides by means of combined aldehyde fuchsin-alcian blue staining. American Journal of Clinical Pathology 33, 453–60.CrossRefGoogle Scholar
Stoward, P. (1968). The histochemical reactions of methanolic thionyl chloride with the anionic groups and basic protein component of sulphated mucosubstances. Journal of the Royal Microscopical Society 88, 119–31.CrossRefGoogle ScholarPubMed
Szirmai, J. A. (1963). Quantitative approaches in the histochemistry of mucopolysaccharides. Journal of Histochemistry and Cytochemistry 11, 24–34.CrossRefGoogle Scholar
Taylor, A. E. R. (1960). Studies on the microfilariae of Loa loa, Wuchereria bancrofti, Brugia malayi, Dirofilaria immitis, D. repens and D. aethiops. Journal of Helminthology 34, 13–26.CrossRefGoogle ScholarPubMed
Thompson, S. W. (1966). Selected Histochemical and Histopathological Methods. Illinois: Thomas.Google Scholar
Urbanova, D. & Adams, C. W. M. (1970). Extraction of lipids from tissue sections with acetone; further qualitative and quantitative histochemical observations. Histochemical Journal 2, 1–9.CrossRefGoogle ScholarPubMed
Wetzel, M. G., Wetzel, B. K. & Spicer, S. S. (1966). Ultrastructural localisation of acid mucosubstances in the mouse colon with iron containing stains. Journal of Cell Biology 30, 299–315.CrossRefGoogle ScholarPubMed