HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) All Versions of this Article: 79/6/1129    most recent biolreprod.108.070177v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal My Folders Download to citation manager Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Vadnais, M. L. Articles by Roberts, K. P. Search for Related Content PubMed PubMed Citation Articles by Vadnais, M. L. Articles by Roberts, K. P. Agricola Articles by Vadnais, M. L. Articles by Roberts, K. P.

Molecular Cloning and Expression of the CRISP Family of Proteins in the Boar¹

Departments of Integrative Biology & Physiology and Urologic Surgery,³ University of Minnesota, Minneapolis, Minnesota 55455 Department of Animal Science,⁴ University of Minnesota, St. Paul, Minnesota 55108

ABSTRACT

The family of mammalian cysteine-rich secretory proteins (CRISP) have been well characterized in the rat, mouse, and human. Here we report the molecular cloning and expression analysis of CRISP1, CRISP2, and CRISP3 in the boar. A partial sequence published in the National Center for Biotechnology Information (NCBI) database was used to derive the full-length sequences for CRISP1 and CRISP2 using rapid amplification of cDNA ends. RT-PCR confirmed the expression of these mRNAs in the boar reproductive tract, and real time RT-PCR showed CRISP1 to be highly expressed throughout the epididymis, with CRISP2 highly expressed in the testis. A search of the porcine genomic sequence in the NCBI database identified a BAC (CH242-199E6) encoding the CRISP1 gene. This BAC is derived from porcine Chromosome 7 and is syntenic with the regions of the mouse, rat, and human genomes encoding the CRISP gene family. This BAC was found to encode a third CRISP protein with a predicted amino acid sequence of high similarity to human CRISP3. Using RT-PCR we show that CRISP3 expression in the boar reproductive tract is confined to the prostate. Recombinant porcine (rp) CRISP2 protein was produced and purified. When incubated with capacitated boar sperm, rpCRISP2 induced an acrosome reaction, consistent with its demonstrated ability to alter the activity of calcium channels.

acrosome reaction, boar, CRISP, epididymis, gamete biology, male reproductive tract, reproductive tract, sperm, testis

¹Supported by the Sorkness Endowment for Research in Urology, Department of Urologic Surgery, University of Minnesota. Porcine CRISP1, CRISP2, and CRISP3 nucleotide sequences were submitted to the National Center for Biotechnology Information and assigned GenBank accession numbers EU661592 (CRISP1), EU661593 (CRISP2), and EU661594 (CRISP3).

²Correspondence and current address: Ken Roberts, WWAMI Medical Education Program, Washington State University, HSB 320N, P.O. Box 1495, Spokane, WA 99210. FAX: 509 358 7627; e-mail: kenroberts{at}wsu.edu.