Cloning and sequence analysis of rat fertilin a and β - Developmental expression, processing and immunolocalization

Eileen A. McLaughlin, Jan Frayne, Helen L. Barker, Jennifer A. Jury, Roy Jones, W. Christopher L. Ford, Len Hall

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Abstract

Fertilin α and β are members of the MDC (metalloproteinase-like, disintegrin-like, cysteine-rich) protein family and are expressed on the sperm surface where they have been proposed to play a role in mammalian fertilization. Inhibition of sperm-oocyte binding and sperm-oocyte fusion make fertilin an attractive target for the development of an immunocontraceptive vaccine. Full-length cDNAs encoding α and β fertilin subunits were isolated from a rat testis cDNA library and sequenced. Using reverse transcription-polymerase chain reaction (RT-PCR), the developmental expression of fertilin α and β was determined in pre-pubertal and mature rat testes. Fertilin α mRNA was present at all stages of development, suggesting that it is not exclusively expressed in post-meiotic germ cells. In contrast, fertilin β mRNA was first identified in day 19 testes, coincident with the presence of pachytene spermatocytes. Polyclonal antisera raised against a 28-residue peptide (corresponding to part of the disintegrin domain) and two recombinant fusion proteins identified a 90 kDa protein in testicular sperm extracts and a 60 kDa protein in caput and cauda epididymidal sperm extracts, the predicted sizes for rat fertilin β precursor and mature protein respectively. Indirect immunofluorescence using the anti-peptide antisera stained the acrosomal cap of permeabilized testicular, caput and caudal spermatozoa and elongating spermatids in testicular sections.

Original languageEnglish
Pages (from-to)801-809
Number of pages9
JournalMolecular Human Reproduction
Volume3
Issue number9
DOIs
Publication statusPublished - 1 Sep 1997
Externally publishedYes

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McLaughlin, E. A., Frayne, J., Barker, H. L., Jury, J. A., Jones, R., Ford, W. C. L., & Hall, L. (1997). Cloning and sequence analysis of rat fertilin a and β - Developmental expression, processing and immunolocalization. Molecular Human Reproduction, 3(9), 801-809. https://doi.org/10.1093/molehr/3.9.801