Reduced curvilinear velocity of boar sperm on substrates with increased hydrophobicity

More suitable and efficient methods to protect gametes from external harmful effects during in vitro handling can be achieved by adding preovulatory porcine oviductal fluid (pOF) to in vitro culture media. The objective of this study was to assess the swim-up procedure’s suitability as a sperm selection method using a medium supplemented with 1mg/mL BSA, 1% preovulatory pOF (v/v), 1% v/v pOF plus 1mg/mL BSA, and 5mg/mL BSA. After selection, various sperm parameters were studied, such as sperm recovery rate, sperm morphology, motility (by CASA), vitality, acrosome status and intracellular calcium (by flow cytometry) and ability to penetrate oocytes in vitro. Around 2% of sperm were recovered after swim-up, and the replacement of BSA by pOF showed a beneficial reduction of motility parameters calcium concentration, resulting in an increased penetration rate. The combination of albumin and oviductal fluid in the medium did not improve the sperm parameters results, whereas a high concentration of BSA increased sperm morphological abnormalities, motility, and acrosome damage, with a reduction of calcium concentration and penetration rate. In conclusion, the replacement of albumin by preovulatory oviductal fluid in the swim-up sperm preparation method modifies boar sperm parameters and improves the in vitro penetration of oocytes.

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Effect of long-term storage in Safe Cell+ extender on boar sperm DNA integrity and other key sperm parameters

Acta Veterinaria Scandinavica ◽

10.1186/s13028-017-0325-9 ◽

2017 ◽

Vol 59 (1) ◽

Cited By ~ 7

Author(s):

Wiesław Bielas ◽

Wojciech Niżański ◽

Agnieszka Partyka ◽

Anna Rząsa ◽

Ryszard Mordak

Keyword(s):

Sperm Parameters ◽

Dna Integrity ◽

Long Term Storage ◽

Sperm Dna Integrity ◽

Sperm Dna ◽

Boar Sperm ◽

Term Storage

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Different Ca2+-releasing abilities of sperm extracts compared with tissue extracts and phospholipase C isoforms in sea urchin egg homogenate and mouse eggs

Biochemical Journal ◽

10.1042/bj3460743 ◽

2000 ◽

Vol 346 (3) ◽

pp. 743-749 ◽

Cited By ~ 32

Author(s):

Keith T. JONES ◽

Miho MATSUDA ◽

John PARRINGTON ◽

Matilda KATAN ◽

Karl SWANN

Keyword(s):

Phospholipase C ◽

Sea Urchin ◽

Tissue Extracts ◽

Sea Urchin Egg ◽

Boar Sperm ◽

Specific Activities ◽

Mammalian Eggs ◽

Mouse Eggs

A soluble phospholipase C (PLC) from boar sperm generates InsP3 and hence causes Ca2+ release when added to sea urchin egg homogenate. This PLC activity is associated with the ability of sperm extracts to cause Ca2+ oscillations in mammalian eggs following fractionation. A sperm PLC may, therefore, be responsible for causing the observed Ca2+ oscillations at fertilization. In the present study we have further characterized this boar sperm PLC activity using sea urchin egg homogenate. Consistent with a sperm PLC acting on egg PtdIns(4,5)P2, the ability of sperm extracts to release Ca2+ was blocked by preincubation with the PLC inhibitor U73122 or by the addition of neomycin to the homogenate. The Ca2+-releasing activity was also detectable in sperm from other species and in whole testis extracts. However, activity was not observed in extracts from other tissues. Moreover recombinant PLCβ1, -γ1, -γ2, -∆1, all of which had higher specific activities than boar sperm extracts, were not able to release Ca2+ in the sea urchin egg homogenate. In addition these PLCs were not able to cause Ca2+ oscillations following microinjection into mouse eggs. These results imply that the sperm PLC possesses distinct properties that allow it to hydrolyse PtdIns(4,5)P2 in eggs.

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Semi-automatic morphometric analysis of normal boar sperm head

Animal Reproduction Science ◽

10.1016/j.anireprosci.2016.03.045 ◽

2016 ◽

Vol 169 ◽

pp. 112 ◽

Cited By ~ 1

Author(s):

A.M. Syanda ◽

G. Boe-Hansen ◽

N. Satake

Keyword(s):

Morphometric Analysis ◽

Sperm Head ◽

Boar Sperm

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Partial purification of a boar sperm membrane protein inducing sperm agglutinating antibody

Bioscience Reports ◽

10.1007/bf01116572 ◽

1990 ◽

Vol 10 (2) ◽

pp. 131-139

Author(s):

Oyewole Adeyemo ◽

E. O. Okegbile ◽

O. O. Olorunsogo

Keyword(s):

Molecular Weight ◽

Membrane Protein ◽

Gel Electrophoresis ◽

Sodium Dodecyl ◽

Gel Filtration ◽

Partial Purification ◽

Boar Sperm ◽

Sperm Membrane ◽

Triton X

For the development of immunological contraception, attention is being concentrated on the possibility of using a sperm membrane antigen. Boar sperm membrane was extracted with triton-X 100 and fractionated by Sephadex G-150 column chromatography. The glycosylated and nonglycosylated portions of protein peaks from the gel filtration were obtained by fractionating on concanavalin A-Sepharose and eluting the bound protein with 0.3 M methyl mannoside. A glycosylated fraction was found to induce sperm agglutinating antibodies in rabbit. The partially purified protein has a molecular weight of 30 kilodaltons, as determined by sodium dodecyl polyaccyrlamide gel electrophoresis. Further work is planned on the histochemical determination of the origin of this protein and species cross-activity of the antibody.

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Zona pellucida-binding of boar sperm acrosin is associated with the N-terminal peptide of the acrosin B-chain (heavy chain)

FEBS Letters ◽

10.1016/0014-5793(90)80881-i ◽

1990 ◽

Vol 265 (1-2) ◽

pp. 51-54 ◽

Cited By ~ 29

Author(s):

E. Töpfer-Petersen ◽

M. Steinberger ◽

C. Ebner von Eschenbach ◽

A. Zucker

Keyword(s):

Heavy Chain ◽

Zona Pellucida ◽

Boar Sperm

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Effects of chelating agents during freeze-drying of boar spermatozoa on DNA fragmentation and on developmental ability in vitro and in vivo after intracytoplasmic sperm head injection

Zygote ◽

10.1017/s0967199406003935 ◽

2007 ◽

Vol 15 (1) ◽

pp. 15-24 ◽

Cited By ~ 38

Author(s):

M. Nakai ◽

N. Kashiwazaki ◽

A. Takizawa ◽

N. Maedomari ◽

M. Ozawa ◽

...

Keyword(s):

Dna Fragmentation ◽

Freeze Drying ◽

Chelating Agents ◽

Sperm Head ◽

Control Group ◽

Sperm Dna Fragmentation ◽

Freeze Dried ◽

Boar Sperm

SUMMARYSuccessful offspring production after intracytoplasmic injection of freeze-dried sperm has been reported in laboratory animals but not in domesticated livestock, including pigs. The integrity of the DNA in the freeze-dried sperm is reported to affect embryogenesis. Release of endonucleases from the sperm is one of the causes of induction of sperm DNA fragmentation. We examined the effects of chelating agents, which inhibit the activation of such enzymes, on DNA fragmentation in freeze-dried sperm and on the in vitro and in vivo developmental ability of porcine oocytes following boar sperm head injection. Boar ejaculated sperm were sonicated, suspended in buffer supplemented with (1) 50 mM EGTA, (2) 50 mM EDTA, (3) 10 mM EDTA, or (4) no chelating agent and freeze-dried. A fertilization medium (Pig-FM) was used as a control. The rehydrated spermatozoa in each group were then incubated in Pig-FM at room temperature. The rate of DNA fragmentation in the control group, as assessed by the TUNEL method, increased gradually as time after rehydration elapsed (2.8% at 0 min to 12.2% at 180 min). However, the rates in all experimental groups (1–4) did not increase, even at 180 min (0.7–4.1%), which were all significantly lower (p < 0.05) than that of the control group. The rate of blastocyst formation after the injection in the control group (6.0%) was significantly lower (p < 0.05) than those in the 50 mM EGTA (23.1%) and 10 mM EDTA (22.6%) groups incubated for 120–180 min. The average number of blastocyst cells in the 50 mM EGTA group (33.1 cells) was significantly higher (p < 0.05) than that in the 10 mM EDTA group (17.8 cells). Finally, we transferred oocytes from 50 mM EGTA or control groups incubated for 0–60 min into estrous-synchronized recipients. The two recipients of the control oocytes became pregnant and one miscarried two fetuses on day 39.The results suggested that fragmentation of DNA in freeze-dried boar sperm is one of the causes of decreased in vitro developmental ability of injected oocytes to the blastocyst stage. Supplementation with EGTA in a freeze-drying buffer improves this ability.

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