Identification and expression analysis of Drosophila melanogaster genes encoding β-hexosaminidases of the sperm plasma membrane

F. Cattaneo, J. Intra, Midori Matsumoto, F. Briani, M. Hoshi, M. E. Perotti

Research output: Contribution to journalArticle

45 Citations (Scopus)

Abstract

Sperm surface β-N-acetylhexosaminidases are among the molecules mediating early gamete interactions in invertebrates and vertebrates, including man. The plasma membrane of Drosophila spermatozoa contains two β-N-acetylhexosaminidases, DmHEXA and DmHEXB, which are required for egg fertilization. Here, we demonstrate that three putative Drosophila melanogaster genes predicted to code for β-N-acetylhexosaminidases, Hexo1, Hexo2, and fdl, are all expressed in the male germ line. fdl codes for a homolog of the α-subunit of the mammalian lysosomal β-N-acetylhexosaminidase Hex A. Hexo1 and Hexo2 encode two homologs of the β-subunit of all known β-N-acetylhexosaminidases, which we have named β1 and β2, respectively. Immunoblot analysis of sperm proteins indicated that the gene products associate in different heterodimeric combinations forming DmHEXA, with an αβ2 structure, and DmHEXB, with a β1 β2 structure. Immunofluorescence demonstrated that all the gene products localized to the sperm plasma membrane. Although none of the genes was testis-specific, fdl was highly and preferentially expressed in the testis, whereas Hexo1 and Hexo2 showed broader tissue expression. Enzyme assays carried out on testis and on a variety of somatic tissues corroborated the results of gene expression analysis. These findings for the first time show the in vivo expression in insects of genes encoding β-N-acetylhexosaminidases, the only molecules so far identified as involved in sperm/egg recognition in this class, whereas in mammals, the organisms where these enzymes have been best studied, only two types of polypeptide chains forming dimeric functional β-N-acetylhexosaminidases are present in Drosophila three different gene products are available that might generate numerous dimeric isoforms.

Original languageEnglish
Pages (from-to)786-800
Number of pages15
JournalGlycobiology
Volume16
Issue number9
DOIs
Publication statusPublished - 2006 Sep

Fingerprint

Hexosaminidases
Gene encoding
Cell membranes
Drosophila melanogaster
Spermatozoa
Genes
Cell Membrane
Testis
Drosophila
Ovum
Insect Genes
Hexosaminidase A
Tissue
Sperm-Ovum Interactions
Molecules
Mammals
Enzyme Assays
Enzymes
Invertebrates
Fertilization

Keywords

  • fdl
  • Fertilization
  • Gamete interactions
  • Hexo2
  • Hexol

ASJC Scopus subject areas

  • Biochemistry

Cite this

Identification and expression analysis of Drosophila melanogaster genes encoding β-hexosaminidases of the sperm plasma membrane. / Cattaneo, F.; Intra, J.; Matsumoto, Midori; Briani, F.; Hoshi, M.; Perotti, M. E.

In: Glycobiology, Vol. 16, No. 9, 09.2006, p. 786-800.

Research output: Contribution to journalArticle

Cattaneo, F. ; Intra, J. ; Matsumoto, Midori ; Briani, F. ; Hoshi, M. ; Perotti, M. E. / Identification and expression analysis of Drosophila melanogaster genes encoding β-hexosaminidases of the sperm plasma membrane. In: Glycobiology. 2006 ; Vol. 16, No. 9. pp. 786-800.
@article{e87e3c3f7e2a45e6a7ecf3a023578ccc,
title = "Identification and expression analysis of Drosophila melanogaster genes encoding β-hexosaminidases of the sperm plasma membrane",
abstract = "Sperm surface β-N-acetylhexosaminidases are among the molecules mediating early gamete interactions in invertebrates and vertebrates, including man. The plasma membrane of Drosophila spermatozoa contains two β-N-acetylhexosaminidases, DmHEXA and DmHEXB, which are required for egg fertilization. Here, we demonstrate that three putative Drosophila melanogaster genes predicted to code for β-N-acetylhexosaminidases, Hexo1, Hexo2, and fdl, are all expressed in the male germ line. fdl codes for a homolog of the α-subunit of the mammalian lysosomal β-N-acetylhexosaminidase Hex A. Hexo1 and Hexo2 encode two homologs of the β-subunit of all known β-N-acetylhexosaminidases, which we have named β1 and β2, respectively. Immunoblot analysis of sperm proteins indicated that the gene products associate in different heterodimeric combinations forming DmHEXA, with an αβ2 structure, and DmHEXB, with a β1 β2 structure. Immunofluorescence demonstrated that all the gene products localized to the sperm plasma membrane. Although none of the genes was testis-specific, fdl was highly and preferentially expressed in the testis, whereas Hexo1 and Hexo2 showed broader tissue expression. Enzyme assays carried out on testis and on a variety of somatic tissues corroborated the results of gene expression analysis. These findings for the first time show the in vivo expression in insects of genes encoding β-N-acetylhexosaminidases, the only molecules so far identified as involved in sperm/egg recognition in this class, whereas in mammals, the organisms where these enzymes have been best studied, only two types of polypeptide chains forming dimeric functional β-N-acetylhexosaminidases are present in Drosophila three different gene products are available that might generate numerous dimeric isoforms.",
keywords = "fdl, Fertilization, Gamete interactions, Hexo2, Hexol",
author = "F. Cattaneo and J. Intra and Midori Matsumoto and F. Briani and M. Hoshi and Perotti, {M. E.}",
year = "2006",
month = "9",
doi = "10.1093/glycob/cwl007",
language = "English",
volume = "16",
pages = "786--800",
journal = "Glycobiology",
issn = "0959-6658",
publisher = "Oxford University Press",
number = "9",

}

TY - JOUR

T1 - Identification and expression analysis of Drosophila melanogaster genes encoding β-hexosaminidases of the sperm plasma membrane

AU - Cattaneo, F.

AU - Intra, J.

AU - Matsumoto, Midori

AU - Briani, F.

AU - Hoshi, M.

AU - Perotti, M. E.

PY - 2006/9

Y1 - 2006/9

N2 - Sperm surface β-N-acetylhexosaminidases are among the molecules mediating early gamete interactions in invertebrates and vertebrates, including man. The plasma membrane of Drosophila spermatozoa contains two β-N-acetylhexosaminidases, DmHEXA and DmHEXB, which are required for egg fertilization. Here, we demonstrate that three putative Drosophila melanogaster genes predicted to code for β-N-acetylhexosaminidases, Hexo1, Hexo2, and fdl, are all expressed in the male germ line. fdl codes for a homolog of the α-subunit of the mammalian lysosomal β-N-acetylhexosaminidase Hex A. Hexo1 and Hexo2 encode two homologs of the β-subunit of all known β-N-acetylhexosaminidases, which we have named β1 and β2, respectively. Immunoblot analysis of sperm proteins indicated that the gene products associate in different heterodimeric combinations forming DmHEXA, with an αβ2 structure, and DmHEXB, with a β1 β2 structure. Immunofluorescence demonstrated that all the gene products localized to the sperm plasma membrane. Although none of the genes was testis-specific, fdl was highly and preferentially expressed in the testis, whereas Hexo1 and Hexo2 showed broader tissue expression. Enzyme assays carried out on testis and on a variety of somatic tissues corroborated the results of gene expression analysis. These findings for the first time show the in vivo expression in insects of genes encoding β-N-acetylhexosaminidases, the only molecules so far identified as involved in sperm/egg recognition in this class, whereas in mammals, the organisms where these enzymes have been best studied, only two types of polypeptide chains forming dimeric functional β-N-acetylhexosaminidases are present in Drosophila three different gene products are available that might generate numerous dimeric isoforms.

AB - Sperm surface β-N-acetylhexosaminidases are among the molecules mediating early gamete interactions in invertebrates and vertebrates, including man. The plasma membrane of Drosophila spermatozoa contains two β-N-acetylhexosaminidases, DmHEXA and DmHEXB, which are required for egg fertilization. Here, we demonstrate that three putative Drosophila melanogaster genes predicted to code for β-N-acetylhexosaminidases, Hexo1, Hexo2, and fdl, are all expressed in the male germ line. fdl codes for a homolog of the α-subunit of the mammalian lysosomal β-N-acetylhexosaminidase Hex A. Hexo1 and Hexo2 encode two homologs of the β-subunit of all known β-N-acetylhexosaminidases, which we have named β1 and β2, respectively. Immunoblot analysis of sperm proteins indicated that the gene products associate in different heterodimeric combinations forming DmHEXA, with an αβ2 structure, and DmHEXB, with a β1 β2 structure. Immunofluorescence demonstrated that all the gene products localized to the sperm plasma membrane. Although none of the genes was testis-specific, fdl was highly and preferentially expressed in the testis, whereas Hexo1 and Hexo2 showed broader tissue expression. Enzyme assays carried out on testis and on a variety of somatic tissues corroborated the results of gene expression analysis. These findings for the first time show the in vivo expression in insects of genes encoding β-N-acetylhexosaminidases, the only molecules so far identified as involved in sperm/egg recognition in this class, whereas in mammals, the organisms where these enzymes have been best studied, only two types of polypeptide chains forming dimeric functional β-N-acetylhexosaminidases are present in Drosophila three different gene products are available that might generate numerous dimeric isoforms.

KW - fdl

KW - Fertilization

KW - Gamete interactions

KW - Hexo2

KW - Hexol

UR - http://www.scopus.com/inward/record.url?scp=33748703846&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33748703846&partnerID=8YFLogxK

U2 - 10.1093/glycob/cwl007

DO - 10.1093/glycob/cwl007

M3 - Article

C2 - 16733265

AN - SCOPUS:33748703846

VL - 16

SP - 786

EP - 800

JO - Glycobiology

JF - Glycobiology

SN - 0959-6658

IS - 9

ER -