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. 1998 Aug 15;102(4):695–703. doi: 10.1172/JCI2545

Expression of aquaporin-4 in fast-twitch fibers of mammalian skeletal muscle.

A Frigeri 1, G P Nicchia 1, J M Verbavatz 1, G Valenti 1, M Svelto 1
PMCID: PMC508931  PMID: 9710437

Abstract

In this study we analyzed the expression of aquaporin-4 (AQP4) in mammalian skeletal muscle. Immunohistochemical experiments revealed that affinity-purified AQP4 antibodies stained selectively the sarcolemma of fast-twitch fibers. By immunogold electron microscopy, little or no intracellular labeling was detected. Western blot analysis showed the presence of two immunopositive bands with apparent molecular masses of 30 and 32 kD specifically present in membrane fraction of a fast-twitch rat skeletal muscle (extensor digitorum longus, EDL) and not revealed in a slow-twitch muscle (soleus). PCR Southern blot experiments resulted in a selective amplification in EDL of a 960-bp cDNA fragment encoding for the full-length rat form of AQP4. Functional experiments carried out on isolated skeletal muscle bundle fibers demonstrated that the osmotic response is faster in EDL than in soleus fibers isolated from the same rat. These results provide for the first time evidence for the expression of an aquaporin in skeletal muscle correlated to a specific fiber-type metabolism. Furthermore, we have analyzed AQP4 expression in skeletal muscle of mdx mice in which a decreased density of orthogonal arrays of particles, a typical morphological feature of AQP4, has been reported. Immunofluorescence experiments showed a marked reduction of AQP4 expression suggesting a critical role in the membrane alteration of Duchenne muscular dystrophy.

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Selected References

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  1. Armstrong R. B., Phelps R. O. Muscle fiber type composition of the rat hindlimb. Am J Anat. 1984 Nov;171(3):259–272. doi: 10.1002/aja.1001710303. [DOI] [PubMed] [Google Scholar]
  2. Berman D. M., Peña-Rasgado C., Holmgren M., Hawkins P., Rasgado-Flores H. External Ca effect on water permeability, regulatory volume decrease, and extracellular space in barnacle muscle cells. Am J Physiol. 1993 Oct;265(4 Pt 1):C1128–C1137. doi: 10.1152/ajpcell.1993.265.4.C1128. [DOI] [PubMed] [Google Scholar]
  3. Farinas J., Simanek V., Verkman A. S. Cell volume measured by total internal reflection microfluorimetry: application to water and solute transport in cells transfected with water channel homologs. Biophys J. 1995 Apr;68(4):1613–1620. doi: 10.1016/S0006-3495(95)80335-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Folkesson H. G., Matthay M. A., Frigeri A., Verkman A. S. Transepithelial water permeability in microperfused distal airways. Evidence for channel-mediated water transport. J Clin Invest. 1996 Feb 1;97(3):664–671. doi: 10.1172/JCI118463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Frigeri A., Gropper M. A., Turck C. W., Verkman A. S. Immunolocalization of the mercurial-insensitive water channel and glycerol intrinsic protein in epithelial cell plasma membranes. Proc Natl Acad Sci U S A. 1995 May 9;92(10):4328–4331. doi: 10.1073/pnas.92.10.4328. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Frigeri A., Gropper M. A., Umenishi F., Kawashima M., Brown D., Verkman A. S. Localization of MIWC and GLIP water channel homologs in neuromuscular, epithelial and glandular tissues. J Cell Sci. 1995 Sep;108(Pt 9):2993–3002. doi: 10.1242/jcs.108.9.2993. [DOI] [PubMed] [Google Scholar]
  7. Han Z., Wax M. B., Patil R. V. Regulation of aquaporin-4 water channels by phorbol ester-dependent protein phosphorylation. J Biol Chem. 1998 Mar 13;273(11):6001–6004. doi: 10.1074/jbc.273.11.6001. [DOI] [PubMed] [Google Scholar]
  8. Hasegawa H., Ma T., Skach W., Matthay M. A., Verkman A. S. Molecular cloning of a mercurial-insensitive water channel expressed in selected water-transporting tissues. J Biol Chem. 1994 Feb 25;269(8):5497–5500. [PubMed] [Google Scholar]
  9. Hatton J. D., Ellisman M. H. Orthogonal arrays are redistributed in the membranes of astroglia from alumina-induced epileptic foci. Epilepsia. 1984 Apr;25(2):145–151. doi: 10.1111/j.1528-1157.1984.tb04170.x. [DOI] [PubMed] [Google Scholar]
  10. Hirche H., Schumacher E., Hagemann H. Extracellular K+ concentration and K+ balance of the gastrocnemius muscle of the dog during exercise. Pflugers Arch. 1980 Sep;387(3):231–237. doi: 10.1007/BF00580975. [DOI] [PubMed] [Google Scholar]
  11. Jung J. S., Bhat R. V., Preston G. M., Guggino W. B., Baraban J. M., Agre P. Molecular characterization of an aquaporin cDNA from brain: candidate osmoreceptor and regulator of water balance. Proc Natl Acad Sci U S A. 1994 Dec 20;91(26):13052–13056. doi: 10.1073/pnas.91.26.13052. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. King L. S., Agre P. Pathophysiology of the aquaporin water channels. Annu Rev Physiol. 1996;58:619–648. doi: 10.1146/annurev.ph.58.030196.003155. [DOI] [PubMed] [Google Scholar]
  13. Kowalchuk J. M., Heigenhauser G. J., Lindinger M. I., Sutton J. R., Jones N. L. Factors influencing hydrogen ion concentration in muscle after intense exercise. J Appl Physiol (1985) 1988 Nov;65(5):2080–2089. doi: 10.1152/jappl.1988.65.5.2080. [DOI] [PubMed] [Google Scholar]
  14. Lindinger M. I., Spriet L. L., Hultman E., Putman T., McKelvie R. S., Lands L. C., Jones N. L., Heigenhauser G. J. Plasma volume and ion regulation during exercise after low- and high-carbohydrate diets. Am J Physiol. 1994 Jun;266(6 Pt 2):R1896–R1906. doi: 10.1152/ajpregu.1994.266.6.R1896. [DOI] [PubMed] [Google Scholar]
  15. Lu M., Lee M. D., Smith B. L., Jung J. S., Agre P., Verdijk M. A., Merkx G., Rijss J. P., Deen P. M. The human AQP4 gene: definition of the locus encoding two water channel polypeptides in brain. Proc Natl Acad Sci U S A. 1996 Oct 1;93(20):10908–10912. doi: 10.1073/pnas.93.20.10908. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Markwell M. A., Haas S. M., Bieber L. L., Tolbert N. E. A modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples. Anal Biochem. 1978 Jun 15;87(1):206–210. doi: 10.1016/0003-2697(78)90586-9. [DOI] [PubMed] [Google Scholar]
  17. Neuhaus J., Schmid E. M., Wolburg H. Stability of orthogonal arrays of particles in murine skeletal muscle and astrocytes after circulatory arrest, and human gliomas. Neurosci Lett. 1990 Feb 5;109(1-2):163–168. doi: 10.1016/0304-3940(90)90556-o. [DOI] [PubMed] [Google Scholar]
  18. Nielsen S., Nagelhus E. A., Amiry-Moghaddam M., Bourque C., Agre P., Ottersen O. P. Specialized membrane domains for water transport in glial cells: high-resolution immunogold cytochemistry of aquaporin-4 in rat brain. J Neurosci. 1997 Jan 1;17(1):171–180. doi: 10.1523/JNEUROSCI.17-01-00171.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Olson J. E., Li G. Z. Increased potassium, chloride, and taurine conductances in astrocytes during hypoosmotic swelling. Glia. 1997 Jul;20(3):254–261. doi: 10.1002/(sici)1098-1136(199707)20:3<254::aid-glia9>3.0.co;2-7. [DOI] [PubMed] [Google Scholar]
  20. Schotland D. L., Bonilla E., Wakayama Y. Freeze fracture studies of muscle plasma membrane in human muscular dystrophy. Acta Neuropathol. 1981;54(3):189–197. doi: 10.1007/BF00687741. [DOI] [PubMed] [Google Scholar]
  21. Seligman A. M., Karnovsky M. J., Wasserkrug H. L., Hanker J. S. Nondroplet ultrastructural demonstration of cytochrome oxidase activity with a polymerizing osmiophilic reagent, diaminobenzidine (DAB). J Cell Biol. 1968 Jul;38(1):1–14. doi: 10.1083/jcb.38.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Shibuya S., Wakayama Y. Changes in muscle plasma membranes in young mice with X chromosome-linked muscular dystrophy: a freeze-fracture study. Neuropathol Appl Neurobiol. 1991 Aug;17(4):335–344. doi: 10.1111/j.1365-2990.1991.tb00730.x. [DOI] [PubMed] [Google Scholar]
  23. Sjøgaard G., Adams R. P., Saltin B. Water and ion shifts in skeletal muscle of humans with intense dynamic knee extension. Am J Physiol. 1985 Feb;248(2 Pt 2):R190–R196. doi: 10.1152/ajpregu.1985.248.2.R190. [DOI] [PubMed] [Google Scholar]
  24. Sorenson A. L. Water permeability of isolated muscle fibers of a marine crab. J Gen Physiol. 1971 Sep;58(3):287–303. doi: 10.1085/jgp.58.3.287. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Suleymanian M. A., Baumgarten C. M. Osmotic gradient-induced water permeation across the sarcolemma of rabbit ventricular myocytes. J Gen Physiol. 1996 Apr;107(4):503–514. doi: 10.1085/jgp.107.4.503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Suzuki M., Iwasaki Y., Yamamoto T., Konno H., Yoshimoto T., Suzuki J. Disintegration of orthogonal arrays in perivascular astrocytic processes as an early event in acute global ischemia. Brain Res. 1984 May 21;300(1):141–145. doi: 10.1016/0006-8993(84)91348-9. [DOI] [PubMed] [Google Scholar]
  27. Tao-Cheng J. H., Bressler J. P., Brightman M. W. Astroglial membrane structure is affected by agents that raise cyclic AMP and by phosphatidylcholine phospholipase C. J Neurocytol. 1992 Jun;21(6):458–467. doi: 10.1007/BF01191509. [DOI] [PubMed] [Google Scholar]
  28. Valenti G., Frigeri A., Ronco P. M., D'Ettorre C., Svelto M. Expression and functional analysis of water channels in a stably AQP2-transfected human collecting duct cell line. J Biol Chem. 1996 Oct 4;271(40):24365–24370. doi: 10.1074/jbc.271.40.24365. [DOI] [PubMed] [Google Scholar]
  29. Verbavatz J. M., Ma T., Gobin R., Verkman A. S. Absence of orthogonal arrays in kidney, brain and muscle from transgenic knockout mice lacking water channel aquaporin-4. J Cell Sci. 1997 Nov;110(Pt 22):2855–2860. doi: 10.1242/jcs.110.22.2855. [DOI] [PubMed] [Google Scholar]
  30. Verkman A. S., van Hoek A. N., Ma T., Frigeri A., Skach W. R., Mitra A., Tamarappoo B. K., Farinas J. Water transport across mammalian cell membranes. Am J Physiol. 1996 Jan;270(1 Pt 1):C12–C30. doi: 10.1152/ajpcell.1996.270.1.C12. [DOI] [PubMed] [Google Scholar]
  31. Wakayama Y., Jimi T., Misugi N., Kumagai T., Miyake S., Shibuya S., Miike T. Dystrophin immunostaining and freeze-fracture studies of muscles of patients with early stage amyotrophic lateral sclerosis and Duchenne muscular dystrophy. J Neurol Sci. 1989 Jun;91(1-2):191–205. doi: 10.1016/0022-510x(89)90087-7. [DOI] [PubMed] [Google Scholar]
  32. Wakayama Y., Kumagai T., Shibuya S. Freeze-fracture studies of muscle plasma membrane in Fukuyama-type congenital muscular dystrophy. Neurology. 1985 Nov;35(11):1587–1593. doi: 10.1212/wnl.35.11.1587. [DOI] [PubMed] [Google Scholar]
  33. Yang B., Brown D., Verkman A. S. The mercurial insensitive water channel (AQP-4) forms orthogonal arrays in stably transfected Chinese hamster ovary cells. J Biol Chem. 1996 Mar 1;271(9):4577–4580. [PubMed] [Google Scholar]
  34. Yang B., Ma T., Verkman A. S. cDNA cloning, gene organization, and chromosomal localization of a human mercurial insensitive water channel. Evidence for distinct transcriptional units. J Biol Chem. 1995 Sep 29;270(39):22907–22913. doi: 10.1074/jbc.270.39.22907. [DOI] [PubMed] [Google Scholar]
  35. Yang B., van Hoek A. N., Verkman A. S. Very high single channel water permeability of aquaporin-4 in baculovirus-infected insect cells and liposomes reconstituted with purified aquaporin-4. Biochemistry. 1997 Jun 17;36(24):7625–7632. doi: 10.1021/bi970231r. [DOI] [PubMed] [Google Scholar]
  36. ZADUNAISKY J. A., PARISI M. N., MONTOREANO R. EFFECT OF ANTIDIURETIC HORMONE ON PERMEABILITY OF SINGLE MUSCLE FIBRES. Nature. 1963 Oct 26;200:365–366. doi: 10.1038/200365a0. [DOI] [PubMed] [Google Scholar]
  37. van Beaumont W., Strand J. C., Petrofsky J. S., Hipskind S. G., Greenleaf J. E. Changes in total plasma content of electrolytes and proteins with maximal exercise. J Appl Physiol. 1973 Jan;34(1):102–106. doi: 10.1152/jappl.1973.34.1.102. [DOI] [PubMed] [Google Scholar]

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