Testosterone could induce a rapid rise in intracellular free Ca2+ concentration through binding to the membrane surface of bone marrow-derived macrophages / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the ways testosterone influences the murine bone marrow-derived macrophages (BMMs) and how testosterone affects the function of BMMs after bound to their membrane surface.</p><p><b>METHODS</b>BMMs were cultured in vitro, their total RNA and proteins isolated, and the expression of intracellular androgen receptor (AR) detected through RT-PCR and Western blotting. The binding site of testosterone (T) to the membrane surface of BMMs was observed by confocal laser scanning microscopy after T-BSA-FITC incubation. Moreover, the intracellular Ca2+ was tested by Fura-2 method, and the influence of ionic currents on BMMs plasma membrane induced by testosterone was examined by the whole cell patch-clamp.</p><p><b>RESULTS</b>RT-PCR and Western blotting failed to detect intracellular ARs in BMMs, but confocal laser scanning microscopy showed testosterone to be bound to the membrane surface of BMMs by impermeable T-BSA-FITC, inducing a rapid rise in the intracellular free Ca2+ concentration ([Ca2+]i) of Fura-2 loaded BMMs, predominantly due to the influx of extracellular Ca2+ through Ni2+ -blockable Ca2+ channels in the plasma membrane. Similarly, the patch-clamp technique revealed T-induced calcium influx in BMMs.</p><p><b>CONCLUSION</b>It is reasonable to assume that the testosterone receptor exists on the plasma membranes, and testosterone act through unconventional plasma membrane receptors, induce Ca2+ influx and a rapid rise in the intracellular Ca2+ concentration, and influence the function of BMMs.</p>

Characterization of the mRNA profile in ejaculated spermatozoa from healthy fertile men / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the complexity of mRNA in the ejaculated sperm from healthy fertile men.</p><p><b>METHODS</b>Semen samples were collected from 10 healthy fathers. The swim-up method was adopted to purify the sperm from possible contamination of somatic cells and the spermatozoal total RNA extracted by Trizol was used for SAGE library analysis.</p><p><b>RESULTS</b>A totle of 21 052 SAGE raw tags were sequenced from 877 clones and 2 712 unique tags that occurred at least twice in the library were given further analysis. 19.7% of the unique tags had no match in the existing SAGE map, representing novel genes. Molecular function analysis revealed 67% of unique tags related to protein binding or nucleic acid binding categories, 41% to catalytic activity, 13% to message transducer activity, and 10% to transporter, structural and transcription regulator activity, respectively.</p><p><b>CONCLUSION</b>There exists a complex repertoire of mRNAs in the ejaculated spermatozoa from fertile men.</p>