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| 1 | Immunomagnetic removal of cryo-damaged human spermatozoa显示文摘Aim: To estimate the dissipation of mitochondrial transmembrane potential (mTMP, △ψm) and activation of sperm caspases (aCP) as signs of apoptosis in human spermatozoa during cryopreservation and to evaluate the efficiency of immunomagnetic cell separation (MACS) of these spermatozoa via annexin V-binding. Methods: The mTMP and aCP in fresh and cryopreserved spermatozoa were detected by fluorescence microscopy and by Western blots. The sperm suspensions were divided into two sperm fractions (with intact and deteriorated membranes) by magnetic cell separation (MiniMACS, Miltenyi Biotec, Bergisch Gladbach, Germany) in dependence on their binding to superparamagnetic annexin V-microbeads (AN-MB). Results: The cryopreservation decreased the portion of spermatozoa with intact mTMP from 80.1% ± 7.2 % to 53.5 % ± 13.1% and increased the spermatozoa with activated pancaspases (aCP) from 21.8 % ± 2.6 % to 47.7 % ± 5.8 % (n = 10; mean ± SEM; P < 0.01). The activation of caspases 1, 3, 8, and 9 in the cryopreserved spermatozoa was confirmed by Western blots (n = 22). MACS reduced significantly the percentage of cryopreserved spermatozoa with dissipated mTMP to 8.1 ± 3.9 (P < 0.01) and also those with aCP to 9.3 % ± 2.2 %. Western blot analyses confirmed the increase of the activated caspase3, 9, and 8 in the AN-MB-positive fraction (P < 0.05) compared with the AN-MB-negative fraction. The MACS separation effect was confirmed by anti-annexin V-antibodies. There was no significant influence of the separation column and the magnetic field on the sperm functions. Conclusion: The cryopreservation impaired the mTMP and enhanced the activation status of caspases in human spermatozoa. The immunomagnetic sperm separation via binding of AN-MB could deplete low quality spermatozoa from cryopreserved semen samples. | Uwe Paasch Sonja Grunewald Katja Wuendrich Tbrsten Jope Hans-Jurgen Glander | 2005 | Asian Journal of Andrology2005，7，1： | 2 |
| 2 | Central role of glycogen synthase kinase-3~ in endoplasmic reticulum stress-induced caspase-3 activation 显示文摘 | Song L De Sarno P Jope RS | 2002 | J Biol Chem2002，277，44： | 1 |
| 3 | Characterization of lithium potentiation of pilocarpine-induced status epilepticus in rats显示文摘 | Jope RS Morrisett RA Shead OC | 1986 | Exp Neurol1986，91，： | 1 |
| 4 | Lithium and GSK-3: one inhibitor, two inhibitory ac- tions, multiple outcomes 显示文摘 | Jope R S | 2003 | Trends Pharmacol Sci2003，24，9： | 1 |
| 5 | Lithium and GSK-3:one inhibitor,two inhibitory ac- tions,multiple outcomes 显示文摘 | Jope RS | 2003 | Trends Pharmacol Sci2003，24，9： | 1 |
| 6 | Inhibition of glycogen synthase kinase-3 protects cells from intrinsic but not extrinsic oxidative stress显示文摘 | King TD Jope RS | 2005 | Neuroreport2005，16，6： | 1 |
| 7 | Proapoptotic stimuli incluce nuclear accumulation of glycogen synthase kinase-3 beta显示文摘 | Bijur GN Jope RS | 2001 | J Biol Chem2001，276，37： | 1 |
| 8 | The role of microtubttle -associated protein 2 (MAP - 2) in neuronal growth,plasticity,and degeneration显示文摘 | Johnson G V Jope R S | 1992 | Neu- rosci Res1992，33，4： | 1 |
| 9 | Glycogen synthase ki- nose-3 (GSK3) : inflammation, diseases, and therapeutics 显示文摘 | Jope RS Yuskaitis CJ Beurel E | 2007 | Neurochem Res2007，32，45： | 1 |
| 10 | Anti-bipolar therapy: mechanism of action of lithium显示文摘 | Jope RS | 1999 | Mol Psychiatry1999，4，2： | 1 |
| 11 | Lithium and GSK-3:one inhibitor, two inhibitory actions, multiple outcomes 显示文摘 | Jope R S | 2003 | Trends Pharmacol Sci2003，24，9： | 1 |
| 12 | Cathepsing is required for sustained inflammation and tissue injury after reperfusion of ischemic kidneys 显示文摘 | Beurel E Jope R | 2006 | Prog Neurobiol2006，79，4： | 1 |
| 13 | Lithium and GSK-3: one inhibitor, two inhibitory actions, multiple outcomes显示文摘 | Jope R S | 2003 | Trends Pharmaeol Sci2003，24，9： | 1 |
| 14 | Glycogen synthase kinase-3 regulates inflammatory tolerance in astrocytes显示文摘 | Beurel E Jope RS | 2010 | Neuroscience2010，169，3： | 1 |
| 15 | Central role of glycogen synthase kinase-3βin endoplasmic reticulum stress-induced caspase-3 activation显示文摘 | Song L De Sarno P Jope RS | 2002 | Journal of Biological Chemistry2002，277，44： | 1 |
| 16 | The muhifaceted roles of glycogen synthase kinase 3beta in cellular signaling 显示文摘 | Grimes C A Jope R S | 2001 | Prog Neurobiol2001，65，4： | 1 |
| 17 | Brain region differences in regulation of Akt and GSK3 by chronic stimulant administration in mice显示文摘 | Mines MA Jope RS | 2012 | Cell Signal2012，24，7： | 1 |
| 18 | Caspase-3 activation induced by inhibition of mitochondrial complex 1 is facilitated by glyco- gen syntbase kinase-3beta and attenuated by lithium 显示文摘 | King T D Bijur G N Jope R S | 2001 | Brain Res2001，919，1： | 1 |
| 19 | AMP-activated protein kinase (AMPK) activating agents cause dephosphorylation of Akt and gly- cogen synthase kinase - 3 显示文摘 | King T D Song L Jope R S | 2006 | Biochem Pharmacol2006，71，11： | 1 |
| 20 | Differential regulation of STAT family me- mbers by glycogen synthase kinase-3 显示文摘 | Beurel E Jope RS | 2008 | J Biol Chem2008，283，21： | 1 |
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