Abstract
Septin14 is an important spermatogenesis related gene involved in the pathogenesis of male infertility that has not been well studied. Here, full-length Septin14 cDNA of the Banna mini-pig inbred line (BMI) was cloned using the RACE method and expressed in pig kidney epithelial cells (PK15) and E. coli Rosetta (DE3) cells. Septin14 expression was identified in somatic tissues and testis in different developmental stages. The pig Septin14 CDS is 1,299 bp long, and encodes a peptide (or protein) of 432 amino acids (MW=50.4 kDa). Phylogenetic analysis indicated that pig Septin14 was highly evolutionarily conserved. Subcellular localization of GFP-tagged Septin14 fusion protein revealed that Septin14 was distributed throughout the testicular cells. Among 34 pig tissues, Septin14 mRNA was found specifically in testis and seminal vesicle. In six different postnatal developmental stages, the testicular level of Septin14 mRNA was barely detectable on day 2, while the highest level occurred on day 75. The spatiotemporal expression profile of Septin14, reported herein for the first time in pig, indicated that Septin14 might be involved in the division, development and apoptosis of germ cells. Furthermore, using a pET prokaryotic expression system, we expressed and isolated recombinant 67.9 kDa Septin14 protein.
Highlights
Septins are guanosine-5-triphosphate (GTP) binding proteins first described in yeast that can self-assemble into polymers and nonpolar oligomers (Mostowy and Cossart, 2012)
We found that Septin14 mRNA was only minimally expressed at day 2 (D2), but was highly expressed by D75, when round spermatids became evident in the cross sections of seminiferous tubules in the Banna mini-pig inbred line (BMI) testes
We reported pig Septin14 gene for the first time
Summary
Septins are guanosine-5-triphosphate (GTP) binding proteins first described in yeast that can self-assemble into polymers and nonpolar oligomers (Mostowy and Cossart, 2012). Septin was first identified in human testis in 2007 (Peterson et al, 2007), and it is highly expressed in testis and is involved in spermatogenesis and cortical neuronal migration (Shinoda et al, 2010). Recent studies reported that the testicular tissues of men with hypo spermatogenesis, maturation arrest and sertoli cells had lower levels of Septin transcripts than normal men Copyright © The Author(s).
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