Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1984 May 1;98(5):1678–1684. doi: 10.1083/jcb.98.5.1678

Diffusion and regionalization in membranes of maturing ram spermatozoa

PMCID: PMC2113169  PMID: 6725394

Abstract

An essential feature of the "fluid mosaic model" (Singer, S. J., and G. L. Nicolson , 1972, Science (Wash. DC)., 175:720-731) of the cell plasma membrane is the ability of membrane lipids and proteins to diffuse laterally in the plane of the membrane. Mammalian sperm are capable of overcoming free random diffusion and restricting specific membrane components, both lipid and protein, to defined regions of the sperm's surface. The patterns of these regionalizations evolve with the processes of sperm differentiation: spermatogenesis, epididymal maturation, and capacitation. We have used the technique of fluorescence recovery after photobleaching to measure the diffusion of the lipid analogue 1,1'- dihexadecyl 3,3,3',3'- tetramethylindocarbocyanine perchlorate ( C16dil ) on the different morphological regions of testicular and ejaculated ram spermatozoa. We have found: (a) that the major morphologically distinct regions (head, midpiece, and tail) of the plasma membrane of both testicular and ejaculated spermatozoa are also physically distinct as measured by C16dil diffusibility; (b) that despite regional differences in diffusibility there is exchange of this lipid analogue by lateral diffusion between the major morphological regions of the plasma membrane; and (c) that epididymal maturation results in changes in C16dil diffusibility in the different regions of the sperm plasma membrane. In particular, the plasma membranes of the anterior and posterior heads become physically distinct.

Full Text

The Full Text of this article is available as a PDF (866.0 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Axelrod D., Koppel D. E., Schlessinger J., Elson E., Webb W. W. Mobility measurement by analysis of fluorescence photobleaching recovery kinetics. Biophys J. 1976 Sep;16(9):1055–1069. doi: 10.1016/S0006-3495(76)85755-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Axelrod D., Ravdin P., Koppel D. E., Schlessinger J., Webb W. W., Elson E. L., Podleski T. R. Lateral motion of fluorescently labeled acetylcholine receptors in membranes of developing muscle fibers. Proc Natl Acad Sci U S A. 1976 Dec;73(12):4594–4598. doi: 10.1073/pnas.73.12.4594. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Barisas B. G., Leuther M. D. Fluorescence photobleaching recovery measurement of protein absolute diffusion constants. Biophys Chem. 1979 Sep;10(2):221–229. doi: 10.1016/0301-4622(79)85044-9. [DOI] [PubMed] [Google Scholar]
  4. Bearer E. L., Friend D. S. Modifications of anionic-lipid domains preceding membrane fusion in guinea pig sperm. J Cell Biol. 1982 Mar;92(3):604–615. doi: 10.1083/jcb.92.3.604. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bradley M. P., Rayns D. G., Forrester I. T. Effects of filipin, digitonin, and polymyxin b on plasma membrane of ram spermatozoa--an EM study. Arch Androl. 1980 May;4(3):195–204. doi: 10.3109/01485018008986963. [DOI] [PubMed] [Google Scholar]
  6. Cherry R. J. Rotational and lateral diffusion of membrane proteins. Biochim Biophys Acta. 1979 Dec 20;559(4):289–327. doi: 10.1016/0304-4157(79)90009-1. [DOI] [PubMed] [Google Scholar]
  7. Dacheux J. L., Voglmayr J. K. Sequence of sperm cell surface differentiation and its relationship to exogenous fluid proteins in the ram epididymis. Biol Reprod. 1983 Nov;29(4):1033–1046. doi: 10.1095/biolreprod29.4.1033. [DOI] [PubMed] [Google Scholar]
  8. Darin-Bennett A., Poulos A., White I. G. The fatty acid composition of the major phosphoglycerides of ram and human spermatozoa. Andrologia. 1976;8(1):37–45. doi: 10.1111/j.1439-0272.1976.tb01643.x. [DOI] [PubMed] [Google Scholar]
  9. Ethier M. F., Wolf D. E., Melchior D. L. Calorimetric investigation of the phase partitioning of the fluorescent carbocyanine probes in phosphatidylcholine bilayers. Biochemistry. 1983 Mar 1;22(5):1178–1182. doi: 10.1021/bi00274a029. [DOI] [PubMed] [Google Scholar]
  10. Frye L. D., Edidin M. The rapid intermixing of cell surface antigens after formation of mouse-human heterokaryons. J Cell Sci. 1970 Sep;7(2):319–335. doi: 10.1242/jcs.7.2.319. [DOI] [PubMed] [Google Scholar]
  11. Johnson L. V., Walsh M. L., Chen L. B. Localization of mitochondria in living cells with rhodamine 123. Proc Natl Acad Sci U S A. 1980 Feb;77(2):990–994. doi: 10.1073/pnas.77.2.990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Klausner R. D., Kleinfeld A. M., Hoover R. L., Karnovsky M. J. Lipid domains in membranes. Evidence derived from structural perturbations induced by free fatty acids and lifetime heterogeneity analysis. J Biol Chem. 1980 Feb 25;255(4):1286–1295. [PubMed] [Google Scholar]
  13. Klausner R. D., Wolf D. E. Selectivity of fluorescent lipid analogues for lipid domains. Biochemistry. 1980 Dec 23;19(26):6199–6203. doi: 10.1021/bi00567a039. [DOI] [PubMed] [Google Scholar]
  14. Koehler J. K. The mammalian sperm surface: studies with specific labeling techniques. Int Rev Cytol. 1978;54:73–108. doi: 10.1016/s0074-7696(08)60165-5. [DOI] [PubMed] [Google Scholar]
  15. Koppel D. E., Axelrod D., Schlessinger J., Elson E. L., Webb W. W. Dynamics of fluorescence marker concentration as a probe of mobility. Biophys J. 1976 Nov;16(11):1315–1329. doi: 10.1016/S0006-3495(76)85776-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Koppel D. E. Fluorescence redistribution after photobleaching. A new multipoint analysis of membrane translational dynamics. Biophys J. 1979 Nov;28(2):281–291. doi: 10.1016/S0006-3495(79)85176-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. McNutt N. S., Weinstein R. S. Membrane ultrastructure at mammalian intercellular junctions. Prog Biophys Mol Biol. 1973;26:45–101. doi: 10.1016/0079-6107(73)90017-5. [DOI] [PubMed] [Google Scholar]
  18. Nicolson G. L., Yanagimachi R. Mobility and the restriction of mobility of plasma membrane lectin-binding components. Science. 1974 Jun 21;184(4143):1294–1296. doi: 10.1126/science.184.4143.1294. [DOI] [PubMed] [Google Scholar]
  19. Pessin J. E., Glaser M. Budding of Rous sarcoma virus and vesicular stomatitis virus from localized lipid regions in the plasma membrane of chicken embryo fibroblasts. J Biol Chem. 1980 Oct 10;255(19):9044–9050. [PubMed] [Google Scholar]
  20. Peters R. Translational diffusion in the plasma membrane of single cells as studied by fluorescence microphotolysis. Cell Biol Int Rep. 1981 Aug;5(8):733–760. doi: 10.1016/0309-1651(81)90231-9. [DOI] [PubMed] [Google Scholar]
  21. Portis A., Newton C., Pangborn W., Papahadjopoulos D. Studies on the mechanism of membrane fusion: evidence for an intermembrane Ca2+-phospholipid complex, synergism with Mg2+, and inhibition by spectrin. Biochemistry. 1979 Mar 6;18(5):780–790. doi: 10.1021/bi00572a007. [DOI] [PubMed] [Google Scholar]
  22. Pringle M. J., Miller K. W. Differential effects on phospholipid phase transitions produced by structurally related long-chain alcohols. Biochemistry. 1979 Jul 24;18(15):3314–3320. doi: 10.1021/bi00582a018. [DOI] [PubMed] [Google Scholar]
  23. Quinn P. J., White I. G. Phospholipid and cholesterol content of epididymal and ejaculated ram spermatozoa and seminal plasma in relation to cold shock. Aust J Biol Sci. 1967 Dec;20(6):1205–1215. doi: 10.1071/bi9671205. [DOI] [PubMed] [Google Scholar]
  24. Schlegel R. A., Phelps B. M., Waggoner A., Terada L., Williamson P. Binding of merocyanine 540 to normal and leukemic erythroid cells. Cell. 1980 Jun;20(2):321–328. doi: 10.1016/0092-8674(80)90618-2. [DOI] [PubMed] [Google Scholar]
  25. Schmell E. D., Yuan L. C., Gulyas B. J., August J. T. Identification of mammalian sperm surface antigens. I. Production of monoclonal anti-mouse sperm antibodies. Fertil Steril. 1982 Feb;37(2):249–257. [PubMed] [Google Scholar]
  26. Scott T. W., Voglmayr J. K., Setchell B. P. Lipid composition and metabolism in testicular and ejaculated ram spermatozoa. Biochem J. 1967 Feb;102(2):456–461. doi: 10.1042/bj1020456. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Shimshick E. J., McConnell H. M. Lateral phase separation in phospholipid membranes. Biochemistry. 1973 Jun 5;12(12):2351–2360. doi: 10.1021/bi00736a026. [DOI] [PubMed] [Google Scholar]
  28. Sims P. J., Waggoner A. S., Wang C. H., Hoffman J. F. Studies on the mechanism by which cyanine dyes measure membrane potential in red blood cells and phosphatidylcholine vesicles. Biochemistry. 1974 Jul 30;13(16):3315–3330. doi: 10.1021/bi00713a022. [DOI] [PubMed] [Google Scholar]
  29. Singer S. J., Nicolson G. L. The fluid mosaic model of the structure of cell membranes. Science. 1972 Feb 18;175(4023):720–731. doi: 10.1126/science.175.4023.720. [DOI] [PubMed] [Google Scholar]
  30. Sklar L. A., Miljanich G. P., Dratz E. A. Phospholipid lateral phase separation and the partition of cis-parinaric acid and trans-parinaric acid among aqueous, solid lipid, and fluid lipid phases. Biochemistry. 1979 May 1;18(9):1707–1716. doi: 10.1021/bi00576a012. [DOI] [PubMed] [Google Scholar]
  31. Staehelin L. A. Structure and function of intercellular junctions. Int Rev Cytol. 1974;39:191–283. doi: 10.1016/s0074-7696(08)60940-7. [DOI] [PubMed] [Google Scholar]
  32. Tank D. W., Wu E. S., Webb W. W. Enhanced molecular diffusibility in muscle membrane blebs: release of lateral constraints. J Cell Biol. 1982 Jan;92(1):207–212. doi: 10.1083/jcb.92.1.207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Voglmayr J. K., Fairbanks G., Vespa D. B., Colella J. R. Studies on mechanisms of surface modifications in ram spermatozoa during the final stages of differentiation. Biol Reprod. 1982 Apr;26(3):483–500. doi: 10.1093/biolreprod/26.3.483. [DOI] [PubMed] [Google Scholar]
  34. Voglmayr J. K., Musto N. A., Saksena S. K., Brown-Woodman D. C., Marley P. B., White I. G. Characteristics of semen collected from the cauda epididymis of conscious rams. J Reprod Fertil. 1977 Mar;49(2):245–251. doi: 10.1530/jrf.0.0490245. [DOI] [PubMed] [Google Scholar]
  35. Voglmayr J. K., Scott T. W., Setchell B. P., Waites G. M. Metabolism of testicular spermatozoa and characteristics of testicular fluid collected from conscious rams. J Reprod Fertil. 1967 Aug;14(1):87–99. doi: 10.1530/jrf.0.0140087. [DOI] [PubMed] [Google Scholar]
  36. Wolf D. E., Edidin M., Handyside A. H. Changes in the organization of the mouse egg plasma membrane upon fertilization and first cleavage: indications from the lateral diffusion rates of fluorescent lipid analogs. Dev Biol. 1981 Jul 15;85(1):195–198. doi: 10.1016/0012-1606(81)90250-5. [DOI] [PubMed] [Google Scholar]
  37. Wolf D. E., Kinsey W., Lennarz W., Edidin M. Changes in the organization of the sea urchin egg plasma membrane upon fertilization: indications from the lateral diffusion rates of lipid-soluble fluorescent dyes. Dev Biol. 1981 Jan 15;81(1):133–138. doi: 10.1016/0012-1606(81)90355-9. [DOI] [PubMed] [Google Scholar]
  38. Wolf D. E., Ziomek C. A. Regionalization and lateral diffusion of membrane proteins in unfertilized and fertilized mouse eggs. J Cell Biol. 1983 Jun;96(6):1786–1790. doi: 10.1083/jcb.96.6.1786. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Wu E. S., Tank D. W., Webb W. W. Unconstrained lateral diffusion of concanavalin A receptors on bulbous lymphocytes. Proc Natl Acad Sci U S A. 1982 Aug;79(16):4962–4966. doi: 10.1073/pnas.79.16.4962. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES