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

Research output: Contribution to journalArticle

37 Citations (Scopus)

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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Sequence Analysis
Organism Cloning
Spermatozoa
Disintegrins
Testis
Oocytes
Immune Sera
Recombinant Fusion Proteins
Messenger RNA
Peptides
Proteins
Spermatocytes
Protein Precursors
Spermatids
Metalloproteases
Indirect Fluorescent Antibody Technique
Fertilins
Gene Library
Fertilization
Germ Cells

Cite this

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
McLaughlin, Eileen A. ; Frayne, Jan ; Barker, Helen L. ; Jury, Jennifer A. ; Jones, Roy ; Ford, W. Christopher L. ; Hall, Len. / Cloning and sequence analysis of rat fertilin a and β - Developmental expression, processing and immunolocalization. In: Molecular Human Reproduction. 1997 ; Vol. 3, No. 9. pp. 801-809.
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Cloning and sequence analysis of rat fertilin a and β - Developmental expression, processing and immunolocalization. / McLaughlin, Eileen A.; Frayne, Jan; Barker, Helen L.; Jury, Jennifer A.; Jones, Roy; Ford, W. Christopher L.; Hall, Len.

In: Molecular Human Reproduction, Vol. 3, No. 9, 01.09.1997, p. 801-809.

Research output: Contribution to journalArticle

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T1 - Cloning and sequence analysis of rat fertilin a and β - Developmental expression, processing and immunolocalization

AU - McLaughlin, Eileen A.

AU - Frayne, Jan

AU - Barker, Helen L.

AU - Jury, Jennifer A.

AU - Jones, Roy

AU - Ford, W. Christopher L.

AU - Hall, Len

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