Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1992 Nov 2;119(4):977–988. doi: 10.1083/jcb.119.4.977

Laminin in the male germ cells of Drosophila

PMCID: PMC2289689  PMID: 1429843

Abstract

To study genes that may be crucial for the male germ cell development of Drosophila we screened a cDNA expression library with a polyclonal antiserum against testis proteins of Drosophila hydei. We identified a cDNA fragment that exhibited a complete sequence similarity with the cDNA of the laminin B2 chain, an important component of the extracellular matrix. Transcripts of laminin B2 were detected in the RNA of male germ cells with the polymerase chain reaction and by in situ hybridization. We studied the reaction of different polyclonal antibodies including those against a Drosophila laminin B2-lac fusion protein, the entire Drosophila laminin complex, or against the mouse laminin complex and against laminin A and B1 chains with specific structures in developing male germ cells of Drosophila. Antigenic sites against laminin B2 were found in the lampbrush loops in primary spermatocyte nuclei, in nuclei of spermatids, and in heads of spermatozoa. The axonemes of elongating spermatids react with antibodies against the Drosophila laminin B1, B2 and laminin A chains. The possible biological functions of the laminin in the male germ cells of Drosophila are discussed.

Full Text

The Full Text of this article is available as a PDF (5.5 MB).

Selected References

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

  1. Blake M. S., Johnston K. H., Russell-Jones G. J., Gotschlich E. C. A rapid, sensitive method for detection of alkaline phosphatase-conjugated anti-antibody on Western blots. Anal Biochem. 1984 Jan;136(1):175–179. doi: 10.1016/0003-2697(84)90320-8. [DOI] [PubMed] [Google Scholar]
  2. Bonaccorsi S., Pisano C., Puoti F., Gatti M. Y chromosome loops in Drosophila melanogaster. Genetics. 1988 Dec;120(4):1015–1034. doi: 10.1093/genetics/120.4.1015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chi H. C., Hui C. F. cDNA and amino acid sequences of Drosophila laminin B2 chain. Nucleic Acids Res. 1988 Jul 25;16(14B):7205–7206. doi: 10.1093/nar/16.14.7205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cooper A. R., MacQueen H. A. Subunits of laminin are differentially synthesized in mouse eggs and early embryos. Dev Biol. 1983 Apr;96(2):467–471. doi: 10.1016/0012-1606(83)90183-5. [DOI] [PubMed] [Google Scholar]
  5. Ekblom M., Klein G., Mugrauer G., Fecker L., Deutzmann R., Timpl R., Ekblom P. Transient and locally restricted expression of laminin A chain mRNA by developing epithelial cells during kidney organogenesis. Cell. 1990 Jan 26;60(2):337–346. doi: 10.1016/0092-8674(90)90748-4. [DOI] [PubMed] [Google Scholar]
  6. Fessler J. H., Fessler L. I. Drosophila extracellular matrix. Annu Rev Cell Biol. 1989;5:309–339. doi: 10.1146/annurev.cb.05.110189.001521. [DOI] [PubMed] [Google Scholar]
  7. Fessler L. I., Campbell A. G., Duncan K. G., Fessler J. H. Drosophila laminin: characterization and localization. J Cell Biol. 1987 Nov;105(5):2383–2391. doi: 10.1083/jcb.105.5.2383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Garzino V., Berenger H., Pradel J. Expression of laminin and of a laminin-related antigen during early development of Drosophila melanogaster. Development. 1989 May;106(1):17–27. doi: 10.1242/dev.106.1.17. [DOI] [PubMed] [Google Scholar]
  9. Gundersen G. G., Shapiro B. M. Sperm surface proteins persist after fertilization. J Cell Biol. 1984 Oct;99(4 Pt 1):1343–1353. doi: 10.1083/jcb.99.4.1343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gültekin H., Heermann K. H. The use of polyvinylidenedifluoride membranes as a general blotting matrix. Anal Biochem. 1988 Aug 1;172(2):320–329. doi: 10.1016/0003-2697(88)90451-4. [DOI] [PubMed] [Google Scholar]
  11. Hemmati-Brivanlou A., Frank D., Bolce M. E., Brown B. D., Sive H. L., Harland R. M. Localization of specific mRNAs in Xenopus embryos by whole-mount in situ hybridization. Development. 1990 Oct;110(2):325–330. doi: 10.1242/dev.110.2.325. [DOI] [PubMed] [Google Scholar]
  12. Hennig W., Kremer H. Spermatogenesis of Drosophila hydei. Int Rev Cytol. 1990;123:129–175. doi: 10.1016/s0074-7696(08)60673-7. [DOI] [PubMed] [Google Scholar]
  13. Hennig W. Untersuchungen zur Struktur und Funktion des Lampenbürsten-Y-Chromosoms in der Spermatogenese von Drosophila. Chromosoma. 1967;22(3):294–357. doi: 10.1007/BF00319879. [DOI] [PubMed] [Google Scholar]
  14. Hennig W. Y chromosome function and spermatogenesis in Drosophila hydei. Adv Genet. 1985;23:179–234. doi: 10.1016/s0065-2660(08)60513-1. [DOI] [PubMed] [Google Scholar]
  15. Hulsebos T. J., Hackstein J. H., Hennig W. Involvement of Y chromosomal loci in the synthesis of Drosophila hydei sperm proteins. Dev Biol. 1983 Nov;100(1):238–243. doi: 10.1016/0012-1606(83)90216-6. [DOI] [PubMed] [Google Scholar]
  16. Hulsebos T. J., Hackstein J. H., Hennig W. Lampbrush loop-specific protein of Drosophila hydei. Proc Natl Acad Sci U S A. 1984 Jun;81(11):3404–3408. doi: 10.1073/pnas.81.11.3404. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Klein G., Langegger M., Timpl R., Ekblom P. Role of laminin A chain in the development of epithelial cell polarity. Cell. 1988 Oct 21;55(2):331–341. doi: 10.1016/0092-8674(88)90056-6. [DOI] [PubMed] [Google Scholar]
  18. Kleinman H. K., Ebihara I., Killen P. D., Sasaki M., Cannon F. B., Yamada Y., Martin G. R. Genes for basement membrane proteins are coordinately expressed in differentiating F9 cells but not in normal adult murine tissues. Dev Biol. 1987 Aug;122(2):373–378. doi: 10.1016/0012-1606(87)90302-2. [DOI] [PubMed] [Google Scholar]
  19. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  20. Lander A. D., Fujii D. K., Reichardt L. F. Purification of a factor that promotes neurite outgrowth: isolation of laminin and associated molecules. J Cell Biol. 1985 Sep;101(3):898–913. doi: 10.1083/jcb.101.3.898. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Laurie G. W., Horikoshi S., Killen P. D., Segui-Real B., Yamada Y. In situ hybridization reveals temporal and spatial changes in cellular expression of mRNA for a laminin receptor, laminin, and basement membrane (type IV) collagen in the developing kidney. J Cell Biol. 1989 Sep;109(3):1351–1362. doi: 10.1083/jcb.109.3.1351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. MEYER G. F., HESS O., BEERMANN W. [Phase specific function structure in spermatocyte nuclei of Drosophila melanogaster and their dependence of Y chromosomes]. Chromosoma. 1961;12:676–716. doi: 10.1007/BF00328946. [DOI] [PubMed] [Google Scholar]
  23. Mecham R. P. Receptors for laminin on mammalian cells. FASEB J. 1991 Aug;5(11):2538–2546. doi: 10.1096/fasebj.5.11.1651264. [DOI] [PubMed] [Google Scholar]
  24. Mierendorf R. C., Percy C., Young R. A. Gene isolation by screening lambda gt11 libraries with antibodies. Methods Enzymol. 1987;152:458–469. doi: 10.1016/0076-6879(87)52054-7. [DOI] [PubMed] [Google Scholar]
  25. Montell D. J., Goodman C. S. Drosophila laminin: sequence of B2 subunit and expression of all three subunits during embryogenesis. J Cell Biol. 1989 Nov;109(5):2441–2453. doi: 10.1083/jcb.109.5.2441. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Montell D. J., Goodman C. S. Drosophila substrate adhesion molecule: sequence of laminin B1 chain reveals domains of homology with mouse. Cell. 1988 May 6;53(3):463–473. doi: 10.1016/0092-8674(88)90166-3. [DOI] [PubMed] [Google Scholar]
  27. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Timpl R. Structure and biological activity of basement membrane proteins. Eur J Biochem. 1989 Apr 1;180(3):487–502. doi: 10.1111/j.1432-1033.1989.tb14673.x. [DOI] [PubMed] [Google Scholar]
  29. von Boxberg Y., Wütz R., Schwarz U. Use of the biotin-avidin system for labelling, isolation and characterization of neural cell-surface proteins. Eur J Biochem. 1990 Jun 20;190(2):249–256. doi: 10.1111/j.1432-1033.1990.tb15569.x. [DOI] [PubMed] [Google Scholar]

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

RESOURCES