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ZFIN ID: ZDB-PUB-110609-34
Immunolocalization of Spetex-1 at the Connecting Piece in Spermatozoa of the Musk Shrew (Suncus murinus)
Kaneko, T., Iwamoto, S., Murayama, E., Kurio, H., Inai, T., Oda, S., and Iida, H.
Date: 2011
Source: Zoological science   28(6): 444-452 (Journal)
Registered Authors: Murayama, Emi
Keywords: Spetex-1, spermatozoa, flagella, Suncus murinus, Tektin4, segmented column
MeSH Terms:
  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Cloning, Molecular
  • Immunoblotting
  • Male
  • Molecular Sequence Data
  • Phylogeny
  • Protein Transport/physiology*
  • Shrews/physiology*
  • Spermatozoa/cytology*
  • Spermatozoa/metabolism*
PubMed: 21627455 Full text @ Zool. Sci.
Spetex-1, which has been isolated by differential display and rat cDNA library screening as a haploid spermatid-specific gene, encodes a protein with two coiled-coil motifs that locates at both the segmented column in the connecting piece and outer dense fibers-affiliated satellite fibrils in rat sperm flagella. Orthologs of Spetex-1 are identified in many animal species, including human, chimpanzee, macaque, cow, dog, African clawed frog, green spotted puffer, and zebrafish. In this study, we used RT-PCR in combination with 5' and 3' RACE (Rapid Amplification of cDNA End) technique to isolate Spetex-1 ortholog of the musk shrew (Suneus murinus), which yielded a full-length Suncus Spetex-1 gene containing an open reading frame of 1,908 base pairs encoding a protein of 636 amino acids with the predicted molecular mass of 72,348 Da. Suncus Spetex-1 has two coiled-coil motifs at 118-184 and 242-276 amino acid residues, which is a characteristic shared by mammalian Spetex-1 proteins. To examine the subcellular localization of Spetex-1 in Suncus spermatozoa, we produced the anti-Suncus Spetex-1 antibody and carried out immunocytochemistry. In spite of that the primary structure of Suncus Spetex-1 is basically similar to that of rat and mouse Spetex-1, confocal laser scanning microscopy and immunoelectron microscopy revealed that Spetex-1 was restricted to the segmented column and capitulum in the connecting piece of Suncus spermatozoa and was not detected in other parts of flagella, suggesting a diversity of Spetex-1 localization in mammalian spermatozoa.