Skip to main content
Log in

Histochemical detection of glycogen and glycoconjugates in the inner ear with modified concanavalin A-horseradish peroxidase procedures

  • Papers
  • Published:
The Histochemical Journal Aims and scope Submit manuscript

Summary

Inner ears from neonatal and adult Mongolian gerbils were examined to determine developmental changes in the content of glycogen and glycoconjugates as shown by histochemical application of the jack bean lectin, concanavalin A (con A). Sections of fixed paraffin-embedded inner ears were stained using the con A-horseradish peroxidase sequence in conjunction with prior treatments including periodate oxidation with or without subsequent reduction and diastase digestion. In adult inner ear, brief periodate oxidation followed by reduction and con A-horseradish peroxidase staining demonstrated abundant glycogen in Deiters' cells and in fibrocytes of the spiral ligament and submacular plaque. This procedure also detected diastase-resistant glycoprotein, probably containing N-linked complex-type saccharides, in the basal and marginal regions of the tectorial membrane and in the otolithic membrane. During morphogenesis and maturation, various cochlear cells showed changes in their glycogen content possibly related to stage-specific energy requirements. Cellular glycogen storage reached adult levels by postnatal day 14. The tectorial membrane gradually acquired con A reactivity during the first postnatal week. Thus, application of modified con A staining procedures has provided further knowledge for comparison with data from previous biochemical and histochemical studies of carbohydrate-rich components in the inner ear.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Bélanger, L. F. (1956) Observations on the development, structure and composition of the cochlea of the rat. Ann. Otol. Rhinol. Laryngol. 65, 1060–73.

    Google Scholar 

  • Duvall, A. J.III. & Hukee, M. J. (1976) Delineation of cochlear glycogen by electron microscopy. Ann. Otol. 85, 234–46.

    Google Scholar 

  • Falbe-Hansen, J. (1967) On glycogen in the cochlear duct of foetuses and young of albino rats. Acta Oto-laryngologica 63, 340–6.

    Google Scholar 

  • Falbe-Hansen, J. & Thomsen, E. (1963) Histochemical studies on glycogen in the cochlea of the guinea pig. Acta Otolaryngol. (Stockh.) 56, 429–36.

    Google Scholar 

  • Gil-Loyzaga, P., Raymond, J. & Gabrion, J. (1985a) Carbohydrates detected by lectins in the vestibular organ. Hear. Res. 18, 269–72.

    Google Scholar 

  • Gil-Loyzaga, P., Gabrion, J. & Uziel, A. (1985b) Lectins demonstrate the presence of carbohydrates in the tectorial, membrane of mammalian cochlea. Hear. Res. 20, 1–8.

    Google Scholar 

  • Gil-Loyzaga, P., Bueno, A. M., Broto, J. P. & Perez, J. P. (1990) Effects of perinatal hypothyroidism in the carbohydrate composition of cochlear tectorial membrane. Hear. Res. 45, 151–6.

    Google Scholar 

  • Hilding, D. A., Bahia, I. & Ginzberg, R. D. (1977) Glycogen in the cochlea during development. Acta Otolaryngol. 84, 12–23.

    Google Scholar 

  • Ishii, D., Takahashi, T. & Balogh, K. (1969) Glycogen in the inner ear after acoustic stimulation. Acta Oto-laryngologica 67, 573–82.

    Google Scholar 

  • Ito, M., Spicer, S. S. & Schulte, B. A. (1993) Immunohistochemical localization of brain type glucose transporter in mammalian inner ear: comparison of development and adult stages. Hear. Res. 71, 230–8.

    Google Scholar 

  • Katsuyama, T. & Spicer, S. S. (1978) Histochemical differentiation of complex carbohydrates with variants of the concanavalin A-horseradish peroxidase method. J. Histochem. Cytochem. 26, 233–50.

    Google Scholar 

  • Katsuyama, T., Ono, K., Nakayama, J. & Kanai, M. (1985) Recent advances in mucosubstance histochemistry. In Gastric Mucus and Mucus Secreting Cells (edited by Kawai, K.) pp. 3–18. Amsterdam: Excepta Medica.

    Google Scholar 

  • Khalkhali-Ellis, Z., Hemming, F. W. & Steel, K. P. (1987) Glycoconjugates of the tectorial membrane. Hear. Res. 25, 185–91.

    Google Scholar 

  • Khan, K. M. & Drescher, D. G. (1990) Proteins of the gelatinous layer of the trout saccular otolithic membrane. Hear. Res. 43, 149–58.

    Google Scholar 

  • Lim, D. J. (1987) Development of the tectorial membrane. Hear. Res. 28, 9–21.

    Google Scholar 

  • Lim, D. J. & Rueda, J. (1990) Distribution of glycoconjugates during cochlear development. A histochemical study. Acta Otolaryngol. (Stockh.) 110, 224–33.

    Google Scholar 

  • Marcus, D. C., Thalmann, R. & Marcus, N. Y. (1978) Respiratory rate and ATP content of stria vascularis of guinea pig in vitro. Laryngoscope 88, 1825–35.

    Google Scholar 

  • Mikol, D. D., Wrabetz, L., Marton, L. S. & Stefansson, K. (1988) Developmental changes in the molecular weights of polypeptides in the human CNS that carry the HNK-1 epitope and bind Phaseolus vulgaris lectins. J. Neurochem. 50, 1924–8.

    Google Scholar 

  • Munyer, P. D. & Schulte, B. A. (1991) Immunohistochemical identification of proteoglycans in gelatinous membranes of cat and gerbil inner ear. Hear. Res. 52, 369–78.

    Google Scholar 

  • Osawa, T. & Tsuji, T. (1987) Fraction and structural assessment of liposaccharides and glycoproteins by use of immobilized lectins. Ann. Rev. Biochem. 56, 21–42.

    Google Scholar 

  • Plotz, E. & Perlman, H. B. (1955) A histochemical study of the cochlea. Laryngoscope 65, 291–312.

    Google Scholar 

  • Prieto, J. J., Rueda, J., Sala, M. L. & Merchan, J. A. (1990a) Lectin staining of saccharides in the normal and hypothyroid developing organ of Corti. Dev. Brain Res. 52, 141–9.

    Google Scholar 

  • Prieto, J. J., Rubio, M. E. & Merchan, J. A. (1990b) Localization of anionic sulfate groups in the tectorial membrane. Hear. Res. 45, 283–94.

    Google Scholar 

  • Qvortrup, K. & Rostgaard, J. (1990) Three-dimensional organization of a transcellular tubulocisternal endoplasmic reticulum in epithelial cells of Reissner's membrane in the guinea pig. Cell Tissue Res. 261, 287–99.

    Google Scholar 

  • Richardson, G. P., Russel, I. J., Duance, V. C. & Bailey, A. J. (1987) Polypeptide composition of the mammalian tectorial membrane. Hear. Res. 25, 45–60.

    Google Scholar 

  • Ross, M. D. (1978) Glycogen accumulation in Reissner's membrane following chemical sympathectomy with 6-hydroxydopamine. Acta Otolaryngol 86, 314–30.

    Google Scholar 

  • Rueda, J. & Lim, D. J. (1988) Possible transient sterociliary adhesion molecules expressed during cochlear development: a preliminary study. In Glycoconjugates in Medicine (edited by Ohyama, M. & Muramatsu, T.) pp. 338–50. Tokyo: Professional Postgraduate Services.

    Google Scholar 

  • Santi, P. A. & Anderson, C. B. (1987) A newly identified surface coat on cochlear hair cells. Hear. Res. 27, 47–65.

    Google Scholar 

  • Shimizu, N. & Kumamoto, T. (1952) A tetra-acetate-Schiff method for polysaccharides in tissue sections. Stain Tech. 27, 97–100.

    Google Scholar 

  • Slepecky, N. & Chamberlain, S. C. (1985) The cell coat of inner ear sensory and supporting cells as demonstrated by ruthenium red. Hear. Res. 17, 281–8.

    Google Scholar 

  • Slepecky, N. B., Cefaratti, L. K. & Yoo, T. J. (1992) Type II and type IX collagen form heterotypic fibers in the tectorial membrane of the inner ear. Matrix 11, 80–6.

    Google Scholar 

  • Spicer, S. S. (1965) Diamine methods for differentiating muco-substances histochemically. J. Histochem. Cytochem. 13, 211–34.

    Google Scholar 

  • Spicer, S. S. & Schulte, B. A. (1992a) Diversity of cell glycocon-jugates shown histochemically: a perspective. J. Histochem. Cytochem. 40, 1–38.

    Google Scholar 

  • Spicer, S. S. & Schulte, B. A. (1992b) Creatine kinase in epithelium of the inner ear. J. Histochem. Cytochem. 40, 185–92.

    Google Scholar 

  • Spicer, S. S. & Schulte, B. A. (1993) Cytologic structures unique to Deiters cells of the cochlea. Anat. Rec. 237, 421–30.

    Google Scholar 

  • Stack, C. R. & Webster, D. B. (1971a) Glycogen content in the outer hair cells of kangaroo rat (D. spectabilis) cochlea prior to and following auditory stimulation. Acta Otolaryngol. (Stockh.) 71, 483–93.

    Google Scholar 

  • Sugiyama, S., Spicer, S. S., Munyer, P. D. & Schulte, B. A. (1991a) Distribution of glycoconjugates in ion transport cells of gerbil inner ear. J. Histochem. Cytochem. 39, 425–34.

    Google Scholar 

  • Sugiyama, S., Spicer, S. S., Munyer, P. D. & Schulte, B. A. (1991b) Histochemical analysis of glycoconjugates in gelatinous Membranes of the gerbil's inner ear. Hear. Res. 55, 263–72.

    Google Scholar 

  • Sugiyama, S., Spicer, S. S. & Schulte, B. A. (1992) Ultrastructural localization and semiquantitative analysis of glycocon-jugates in the tectorial membrane. Hear. Res. 58, 35–46.

    Google Scholar 

  • Suzuki, H., Lee, Y. C., Tachibana, M., Hozawa, K., Wataya, H. & Takasaka, T. (1992) Quantitative carbohydrate analyses of the tectorial and otoconial membranes of the guinea pig. Hear. Res. 60, 45–52.

    Google Scholar 

  • Thalmann, I., Thallinger, G., Crouch, E. C., Comegys, T. H., Barrett, N. & Thalmann, R. (1987) Composition and supramolecular organization of the tectorial membrane. Laryngoscope 97, 357–67.

    Google Scholar 

  • Wislocki, G. H. & Ladman, A. J. (1955) Selective and histo-chemical staining of the otolithic membranes, cupulae and tectorial membrane of the inner ear. J. Anat. (London) 89, 3–15.

    Google Scholar 

  • Yamashita, H. & Bagger-Sjöbäck, D. (1992) Expression of glycoconjugates in the human fetal cochlea. Acta Otolaryngol. (Stockh.) 112, 628–34.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ito, M., Spicer, S.S. & Schulte, B.A. Histochemical detection of glycogen and glycoconjugates in the inner ear with modified concanavalin A-horseradish peroxidase procedures. Histochem J 26, 437–446 (1994). https://doi.org/10.1007/BF00160057

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00160057

Keywords

Navigation