Biochemical maturation of Spam1 (PH-20) during epididymal transit of mouse sperm involves modifications ofN-linked oligosaccharides

Deng, Xiaolong; Czymmek, Kirk J.; Martin‐DeLeon, Patricia A.

doi:10.1002/(sici)1098-2795(199902)52:2<196::aid-mrd11>3.0.co;2-a

Cited by 49 publications

(81 citation statements)

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“…Implantation of tumor cells possessing high hyaluronidase activity induced angiogenesis in the cornea of mice that was inhibited by the hyaluronidase inhibitor apigenin. 22 Furthermore, it has been shown that commercial preparations of testicular hyaluronidases, also known as sperm adhesion molecule 1 (SPAM1) or PH-20 protein, [25][26][27] exhibited beneficial effects in myocardial infarction 28 and liver transplantation (V. Papanicolaou, et al, in preparation). These observations prompted us to investigate the effects of testicular hyaluronidase on endothelial cell morphogenesis.…”

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confidence: 99%

Testicular hyaluronidase induces tubular structures of endothelial cells grown in three‐dimensional colagen gel through a CD44‐mediated mechanism

Rahmanian

Heldin

2001

Intl Journal of Cancer

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Cultured brain capillary endothelial cells grown in a 3-dimensional collagen gel can form tubular structures after stimulation by angiogenic factors. We found that treatment of such cultures with testicular hyaluronidase led to formation of tubular structures and cell survival. Anion-exchange chromatography of the enzyme preparation on a MonoQ column revealed the presence of the angiogenic factor basic fibroblast growth factor (bFGF) in the flow-through fraction, as determined by immunoblotting; part of the effect on endothelial cell morphogenesis could thus be ascribed to bFGF. However, adsorbed fractions eluted with increasing concentrations of NaCl, which exhibited hyaluronan-degrading activity at neutral pH, did not contain bFGF but were still able to induce tube-like structures of the endothelial cells. Streptomyces hyaluronidase failed to evoke the same effect. Interestingly, blocking of hyaluronan binding to CD44 receptors by the monoclonal antibody KM114 inhibited the effect of hyaluronidase, but not of bFGF, on endothelial cell tube formation. Our data suggest a CD44-mediated specific role for certain populations of testicular hyaluronidase in the induction of angiogenesis.

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confidence: 99%

Testicular hyaluronidase induces tubular structures of endothelial cells grown in three‐dimensional colagen gel through a CD44‐mediated mechanism

Rahmanian

Heldin

2001

Intl Journal of Cancer

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“…Data are presented as mean ± SE. Pearson's correlation coefficient was used to correlate sperm DNA damage and histone retention with the percentage of negatively (NCS) and positively (PCS) charged sperm at different electrophoretic currents (6,8,10,12, and 14 mA). To identify the effect of PCS and NCS population on the test parameters, a correction for neutrally charged sperm was performed.…”

Section: Discussionmentioning

confidence: 99%

“…During electrophoresis, sperm were assessed for their morphology, and sperm with normal morphology and negatively charge were selected using the ICSI pipette. Subsequently, negatively charged sperm were selected at low current (6,8, and 10 mA) and compared with PCS and controls for DNA damage. Fifty charged sperm were collected within 3-5 min of electrophoretic exposure.…”

Section: Microelectrophoresis Sperm Separationmentioning

confidence: 99%

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Optimization of microelectrophoresis to select highly negatively charged sperm

Lee¹,

Murphy²,

Aston³

et al. 2016

J Assist Reprod Genet

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Purpose The sperm membrane undergoes extensive surface remodeling as it matures in the epididymis. During this process, the sperm is encapsulated in an extensive glycocalyx layer, which provides the membrane with its characteristic negative electrostatic charge. In this study, we develop a method of microelectrophoresis and standardize the protocol to isolate sperm with high negative membrane charge. Methods Under an electric field, the percentage of positively charged sperm (PCS), negatively charged sperm (NCS), and neutrally charged sperm was determined for each ejaculate prior to and following density gradient centrifugation (DGC), and evaluated for sperm DNA damage, and histone retention. Subsequently, PCS, NCS, and neutrally charged sperm were selected using an ICSI needle and directly analyzed for DNA damage. Results When raw semen was analyzed using microelectrophoresis, 94 % were NCS. In contrast, DGC completely or partially stripped the negative membrane charge from sperm resulting PCS and neutrally charged sperm, while the charged sperm populations are increased with an increase in electrophoretic current. Following DGC, high sperm DNA damage and abnormal histone retention were inversely correlated with percentage NCS and directly correlated with percentage PCS. NCS exhibited significantly lower DNA damage when compared with control (P < 0.05) and PCS (P < 0.05). When the charged sperm population was corrected for neutrally charged sperm, sperm DNA damage was strongly associated with NCS at a lower electrophoretic current. Conclusion The results suggest that selection of NCS at lower current may be an important biomarker to select healthy sperm for assisted reproductive treatment.

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“…Following centrifugation, samples were processed immediately as uncapacitated sperm, or either incubated for 90-120 min in HTF medium at 37°C to undergo capacitation, or treated with 1 μM A23187 calcium ionophore and incubated at 37°C for 65 min to undergo the acrosome reaction (AL-Dossary, Bathala, Caplan, & Martin-DeLeon, 2015;Deng, Czymmek, & Martin-DeLeon, 1999). …”

Section: Sperm Collection and Protein Extractionmentioning

confidence: 99%

Plasma membrane calcium ATPase 4 (PMCA4) co‐ordinates calcium and nitric oxide signaling in regulating murine sperm functional activity

Olli

Galileo

et al. 2017

Journal Cellular Physiology

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Reduced sperm motility (asthenospermia) and resulting infertility arise from deletion of the Plasma Membrane Ca 2+ -ATPase 4 (Pmca4) gene which encodes the highly conserved Ca 2+ efflux pump, PMCA4. This is the major Ca 2+ clearance protein in murine sperm. Since the mechanism underlying asthenospermia in PMCA4's absence or reduced activity is unknown, we investigated if sperm PMCA4 negatively regulates nitric oxide synthases (NOSs) and when absent NO, peroxynitrite, and oxidative stress levels are increased. Using co-immunoprecipitation (Co-IP) and Fluorescence Resonance Energy Transfer (FRET), we show an association of PMCA4 with the NOSs in elevated cytosolic [Ca 2+ ] in capacitated and Ca 2+ ionophore-treated sperm and with neuronal (nNOS) at basal [Ca 2+ ] (ucapacitated sperm). FRET efficiencies for PMCA4-eNOS were 35% and 23% in capacitated and uncapacitated sperm, significantly (p < 0.01) different, with the molecules being <10 nm apart. For PMCA4-nNOS, this interaction was seen only for capacitated sperm where FRET efficiency was 24%, significantly (p < 0.05) higher than in uncapacitated sperm (6%). PMCA4 and the NOSs were identified as interacting partners in a quaternary complex that includes Caveolin1, which co-immunoprecipitated with eNOS in a Ca 2+ -dependent manner. In Pmca4 −/− sperm NOS activity was elevated twofold in capacitated/uncapacitated sperm (vs. wild-type), accompanied by a twofold increase in peroxynitrite levels and significantly (p < 0.001) increased numbers of apoptotic germ cells. The data support a quaternary complex model in which PMCA4 co-ordinates Ca 2+ and NO signaling to maintain motility, with increased NO levels resulting in asthenospermia in Pmca4 −/− males.They suggest the involvement of PMCA4 mutations in human asthenospermia, with diagnostic relevance.asthenospermia, calcium efflux pump, infertility, oxidative stress, sperm damageCapacitation is the series of changes that occur in the final maturation of mammalian sperm in the female in order for them to gain fertilizing competence, and subsequently undergo the acrosome reaction. Both of these processes are dependent on elevated levels of cytosolic Ca 2+ concentration ([Ca 2+ ] c ) (de Lamirande, Leclerc, & Gagnon, 1997;Fraser, 1987). After the calcium influx required to meet this high calcium demand, there needs to be a return to resting cytosolic calcium concentration ([Ca 2+ ] c ) levels (50-100 nM [Herrick et al., 2005]). In mice, in the absence of the major calcium efflux pump, Plasma Membrane Calcium ATPase 4 (PMCA4) (Wennemuth, Babcock, & Hille, 2003), calcium levels remain elevated with loss of sperm motility (Asthenospermia, AS) and resulting male infertility (Okunade et al., 2004;Schuh et al., 2004 (de Lamirande et al., 1997;Fraser, 1987) which rapidly activates them (Knowles & Moncada, 1994). Thus, as PMCA4 extrudes Ca 2+ , the NOSs are held in a locale or microdomain with a relatively low [Ca 2+ ] c, compared to the surrounding environment where it is globally high. In this Ca 2+ -extruding locale, NO...

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Biochemical maturation of Spam1 (PH-20) during epididymal transit of mouse sperm involves modifications ofN-linked oligosaccharides

Cited by 49 publications

References 29 publications

Testicular hyaluronidase induces tubular structures of endothelial cells grown in three‐dimensional colagen gel through a CD44‐mediated mechanism

Testicular hyaluronidase induces tubular structures of endothelial cells grown in three‐dimensional colagen gel through a CD44‐mediated mechanism

Optimization of microelectrophoresis to select highly negatively charged sperm

Plasma membrane calcium ATPase 4 (PMCA4) co‐ordinates calcium and nitric oxide signaling in regulating murine sperm functional activity

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