Proteomic characterization of canine seminal plasma

Annice Aquino-Cortez; Breno Queiroz Pinheiro; David Baruc Cruvinel Lima; Herlon Victor Rodrigues Silva; Antônio Cavalcante Mota-Filho; Jorge André Matias Martins; Paula Rodriguez-Villamil; Arlindo Alencar Moura; Lúcia Daniel Machado Silva

doi:10.1016/j.theriogenology.2017.03.016

Proteomic characterization of canine seminal plasma

Theriogenology. 2017 Jun:95:178-186. doi: 10.1016/j.theriogenology.2017.03.016. Epub 2017 Mar 20.

Authors

Annice Aquino-Cortez¹, Breno Queiroz Pinheiro¹, David Baruc Cruvinel Lima¹, Herlon Victor Rodrigues Silva¹, Antônio Cavalcante Mota-Filho², Jorge André Matias Martins³, Paula Rodriguez-Villamil⁴, Arlindo Alencar Moura⁴, Lúcia Daniel Machado Silva⁵

Affiliations

¹ Laboratory of Carnivores Reproduction, State University of Ceara, Fortaleza, Brazil.
² Faculty Earth Northeast, Caucaia, Brazil.
³ Academic Unit of Serra Talhada, Federal Rural University of Pernambuco, Serra Talhada, Brazil.
⁴ Department of Animal Science, Federal University of Ceara, Fortaleza, Brazil.
⁵ Laboratory of Carnivores Reproduction, State University of Ceara, Fortaleza, Brazil. Electronic address: lucia.daniel.machado@hotmail.com.

PMID: 28460673
DOI: 10.1016/j.theriogenology.2017.03.016

Abstract

The present study was conducted to identify the major proteome of the sperm-rich fraction and prostatic fraction of canine seminal plasma. Three semen samples from four healthy dogs were obtained by digital manipulation. The pre-sperm fraction, sperm-rich fraction and prostatic fraction were separated from each ejaculate. Immediately after sperm analysis, a protease inhibitor was added to the sperm-rich fraction and prostatic fraction, and the fractions were separately centrifuged and frozen at -80 °C. The samples were thawed, re-centrifuged, and the total protein concentration was determined. Samples were subjected to 1D SDS-PAGE and Coomassie-blue stained gels, were analyzed by Quantity One 1D Analysis Software. Bands detected in the gels were excised and proteins subjected to digestion with trypsin. Proteins were identified by nano-HPLC-MS and tools of bioinformatics. Tandem mass spectrometry allowed the detection of 268 proteins in the gels of sperm-rich fraction and prostatic fraction of canine ejaculate. A total of 251 proteins were common to the sperm-rich and prostatic fractions, while 17 proteins were present in the sperm-rich fraction and absent in the prostatic fraction. The intensity of the bands detected in range 1 and 2 represented 46.5% of all of the band intensities detected in the 1D gels for proteins of the sperm-rich fraction and 53.0% of all bands in the prostatic fraction. Arginine esterase and lactotransferrin precursor were the protein with the highest intensity observed in the both fractions. Among the proteins present only in the sperm-rich fraction, the proteins UPF0764 protein C16orf89 homolog and epididymal-specific lipocalin-9 were the most abundant. In conclusion, canine sperm-rich fraction and prostatic fraction express a very diverse set of proteins, with unique biochemical properties and functions. Moreover, although most proteins are common to both sperm-rich fraction and prostatic fraction, there are some exclusive proteins in sperm-rich fraction.

Keywords: Canine semen; Dog; Prostatic fraction; Proteins; Proteomic; Sperm-rich fraction.

MeSH terms

Animals
Dogs / physiology*
Male
Proteome / analysis*
Semen / chemistry*
Semen / physiology
Semen Analysis / veterinary*
Seminal Plasma Proteins / analysis*

Substances

Proteome
Seminal Plasma Proteins