Skip to main content
Log in

Carbonic anhydrase isoform expression and functional role in rodent extraocular muscle

  • Skeletal Muscle
  • Published:
Pflügers Archiv Aims and scope Submit manuscript

Abstract

Carbonic anhydrase (CA) accelerates contractile function, particularly in fast-twitch skeletal muscles. Since the extraocular muscles are considered to be amongst the fastest skeletal muscles in mammals, this study tested two hypotheses: (1) CA is expressed at higher levels in rat extraocular muscles than in extensor digitorum longus (EDL, a fast limb muscle), and (2) inhibition of CA activity increases twitch duration and force in the extraocular muscles to a greater extent than in EDL. By real-time quantitative PCR we determined that the expression of CA3 isoform, typically high in skeletal muscles, is significantly depressed in extraocular muscles. Message levels for the CA2 and CA4 isoforms were higher in the extraocular muscles, while CA5 expression was equivalent in both muscles. Strong CA activity was demonstrated by histochemistry in frozen EDL muscle sections, in particular along the sarcolemma and in capillaries. By contrast, extraocular muscle had very low sarcolemmal or cytosolic CA activity. CA inhibition with 6-ethoxyzolamide (ETZ) reversibly increased twitch duration and force in EDL muscle bundles. In the extraocular muscles, ETZ did not alter twitch kinetics. Based on these results, we reject our initial hypotheses and conclude that CA does not influence the fast contractile kinetics characteristic of the extraocular muscles.

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.

Institutional subscriptions

Fig. 1A, B
Fig. 2A, B
Fig. 3

Similar content being viewed by others

References

  1. Andrade FH, Anzueto A, Napier W, Levine S, Lawrence RA, Jenkinson SG, Maxwell LC (1998) Effects of selenium deficiency on diaphragmatic function after resistive loading. Acta Physiol Scand 162:141–148

    Article  CAS  PubMed  Google Scholar 

  2. Asmussen G, Gaunitz U (1981) Mechanical properties of the isolated inferior oblique muscle of the rabbit. Pflugers Archiv 392:183–190

    CAS  PubMed  Google Scholar 

  3. Close RI, Luff AR (1974) Dynamic properties of inferior rectus muscle of the rat. J Physiol (Lond) 236:259–270

    Google Scholar 

  4. Fischer MD, Gorospe JR, Felder E, Bogdanovich S, Pedrosa-Domellöf F, Ahima RS, Rubinstein NA, Hoffman EP, Khurana TS (2002) Expression profiling reveals metabolic and structural components of extraocular muscles. Physiol Genomics 9:71–84

    Google Scholar 

  5. Fuchs AF, Scudder CA, Kaneko CRS (1988) Discharge patterns and recruitment order of identified motoneurons and internuclear neurons in the monkey abducens nucleus. J Neurophysiol 60:1874–1895

    CAS  PubMed  Google Scholar 

  6. Geers C, Gros G (2000) Carbon dioxide transport and carbonic anhydrase in blood and muscle. Physiol Rev 80:681–715

    CAS  PubMed  Google Scholar 

  7. Giulietti A, Overbergh L, Valckx D, Decallonne B, Bouillon R, Mathieu C (2001) An overview of real-time quantitative PCR: applications to quantify cytokine gene expression. Methods 25:386–401

    Article  CAS  PubMed  Google Scholar 

  8. Hansson HPJ (1967) Histochemical demonstration of carbonic anhydrase activity. Histochemie 11:112–128

    CAS  PubMed  Google Scholar 

  9. Hennig R, Lømo T (1985) Firing patterns of motor units in normal rats. Nature 314:164–166

    CAS  PubMed  Google Scholar 

  10. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2ΔΔCt method. Methods 25:402–408

    CAS  PubMed  Google Scholar 

  11. Mayr R (1971) Structure and distribution of fibre types in the external eye muscles of the rat. Tissue Cell 3:433–462

    Google Scholar 

  12. Peyronnard J-M, Charron LF, Messier J-P, Lavoie J, Faraco-Cantin F, Dubreuil M (1988) Histochemical localization of carbonic anhydrase in normal and disease human muscle. Muscle Nerve 11:108–113

    CAS  PubMed  Google Scholar 

  13. Spencer RF, Porter JD (1988) Structural organization of the extraocular muscles. In: Büttner-Ennever JA (ed) Neuroanatomy of the oculomotor system. Elsevier, New York, pp 33–79

  14. Spitzer KW, Skolnick RL, Peercy BE, Keener JP, Vaughan-Jones RD (2002) Facilitation of intracellular H+ ion mobility by CO2/HCO3 in rabbit ventricular myocytes is regulated by carbonic anhydrase. J Physiol (Lond) 541:159–167

    Google Scholar 

  15. Wetzel P, Gros G (1998) Inhibition and kinetic properties of membrane-bound carbonic anhydrases in rabbit skeletal muscles. Arch Biochem Biophys 356:151–158

    CAS  PubMed  Google Scholar 

  16. Wetzel P, Kleinke T, Papadopoulos S, Gros G (2002) Inhibition of muscle carbonic anhydrase slows the Ca2+ transient in rat skeletal muscle fibers. Am J Physiol 283:C1242–C1253

    CAS  Google Scholar 

  17. Wetzel P, Papadopoulos S, Gros G (2002) Inhibition of muscle carbonic anhydrase increases rise and relaxation times of twitches in rat skeletal muscle fibres. Pflugers Arch 443:762–770

    Article  CAS  PubMed  Google Scholar 

  18. Wieczorek DF, Periasamy M, Butler-Browne GS, Whalen RG, Nadal-Ginard B (1985) Co-expression of multiple myosin heavy chain genes, in addition to a tissue-specific one, in extraocular musculature. J Cell Biol 101:618–629

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank the Center for AIDS Research at CWRU (P30 AI36219) for providing access to real-time quantitative PCR. This work was supported by grants from the National Eye Institute (EY12998 and EY13724 to F.H.A., and P30 EY11373 to the Vision Science Research Center at CWRU) and by the Evenor Armington Fund.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Francisco H. Andrade.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Andrade, F.H., Hatala, D.A. & McMullen, C.A. Carbonic anhydrase isoform expression and functional role in rodent extraocular muscle. Pflugers Arch - Eur J Physiol 448, 547–551 (2004). https://doi.org/10.1007/s00424-004-1284-3

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00424-004-1284-3

Keywords

Navigation