OSR1 and SPAK cooperatively modulate Sertoli cell support of mouse spermatogenesis

Abstract

We investigated the role of oxidative stress-responsive kinase-1 (OSR1) and STE20 (sterile 20)/SPS1-related proline/alanine-rich kinase (SPAK), upstream regulators of the Na+-K+-2Cl− cotransporter (NKCC1)—essential for spermatogenesis—in mouse models of male fertility. Global OSR1+/− gene mutations, but not global SPAK−/− or Sertoli cell (SC)-specific OSR1 gene knockout (SC-OSR1−/−), cause subfertility with impaired sperm function and are associated with reduced abundance of phosphorylated (p)-NKCC1 but increased p-SPAK expression in testicular tissue and spermatozoa. To dissect further in a SC-specific manner the compensatory effect of OSR1 and SPAK in male fertility, we generated SC-OSR1−/− and SPAK−/− double knockout (DKO) male mice. These are infertile with defective spermatogenesis, presenting a SC-only-like syndrome. Disrupted meiotic progression and increased germ cell apoptosis occurred in the first wave of spermatogenesis. The abundance of total and p-NKCC1 was significantly decreased in the testicular tissues of DKO mice. These results indicate that OSR1 and SPAK cooperatively regulate NKCC1-dependent spermatogenesis in a SC-restricted manner.

hnRNPU in Sertoli cells cooperates with WT1 and is essential for testicular development by modulating transcriptional factors Sox8/9

“…The results advised that there were 19 distinctly differential transcription factors between Amh-Cre-cKO and control testes at P3 (Figure 6 D) . We then chose 3 transcription factors associated with Sertoli cell and testis development, including Rhox8 44 , Lhx1 45 , and Osr1 46 , to validate the RNA-seq data. The RT-qPCR results were in keeping with the RNA-seq data (Figure 6 E-F) .…”

Loss of Slc12a2 specifically in pancreatic β-cells drives metabolic syndrome in mice

“…Indeed, i ) Cre -mediated recombination of "floxed" alleles was only detected in β-cells of Ins1 Cre : Nkcc1 lox/lox : Tomato ( Fig 1A–1H ) and Nkcc1 βKO mice ( Fig 1I–1L ), respectively; ii ) Nkcc1 βKO islets exhibited expected recombination events and Nkcc1 transcript expression patterns ( Fig 1M–1P ); and iii ) immunoreactive Cre was present only in β-cells of Ins1 Cre mice ( S2I–S2P Fig ). Importantly, the use of Nkcc1 antibodies validated against knockout tissues ( S1A–S1H Fig ) showed the expected expression pattern of Nkcc1 in tissues/cells of the normal Nkcc1 lox/lox mouse, including those in Sertoli and chromaffin cells ( S2Q and S2R Fig ) [ 70 , 71 ], ductal/epithelial cells of the pancreas, intestine ( S2S and S2T Fig ) [ 72 , 73 ] and brain in Ins1 Cre : Nkcc1 lox/lox : Tomato mouse model ( S1E–S1I Fig ) [ 74 ]. Along the same lines, Nkcc1 protein expression was intact in the choroid plexus ( S1I Fig ), a relevant finding because low levels of Ins1 gene activity were previously reported [ 75 ].…”

The Ubiquitin-Proteasome System Participates in Sperm Surface Subproteome Remodeling during Boar Sperm Capacitation

“…The STK39 is expressed at high levels in normal human testes and the prostate [ 72 ], as well as in several cancer cell lines [ 72 , 73 , 110 , 111 , 112 , 113 ]. The importance of STK39 in mouse spermatogenesis was demonstrated by Liu et al [ 71 ], where a Sertoli cell-specific OSR1 −/− and STK39 −/− double-knockout male mice displayed infertility with increased germ cell apoptosis and defective spermatogenesis, resulting in Sertoli cell-only syndrome. These double-knockout male mice had a significantly decreased abundance of phosphorylated NKCC1 (the ubiquitous form, which is more active than dephosphorylated form), suggesting that STK39, in cooperation with OSR1, regulates spermatogenesis via the activation of NKCC1.…”