Motility of Bovine Spermatozoa Extended in “Defined” Diluents

Gebauer, Matthias; Pickett, B.W.; Komarek, R.J.; Gaunya, W.S.

doi:10.3168/jds.s0022-0302(70)86297-x

Cited by 25 publications

(21 citation statements)

References 16 publications

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“…Although the mechanisms by which EY protects sperm during low-temperature storage are unclear, its low-density lipoprotein fraction (LDF) has been identified as the active constituent [35][36][37]. Although it has been reported that exogenous phospholipids protect sperm against cold shock [28,[38][39][40][41], EY or EY lipoprotein is more effective for protecting sperm during both cold storage [29] and freezing and thawing [31,42]. The superiority of lipoprotein over phospholipids prepared from EY has been interpreted as evidence for the presence of two distinct factors [28,29].…”

Section: Effect Of Ey On Survival Of Mouse Sperm After Freezing and Tmentioning

confidence: 99%

Birth of Live Mice Resulting from Oocytes Fertilized in Vitro with Cryopreserved Spermatozoa1

Songsasen

Betteridge

1997

Biology of Reproduction

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The objective of this study was to develop a reliable method to cryopreserve mouse spermatozoa, using in vitro fertilization (IVF) of oocytes, development of resultant zygotes into blastocysts, and the birth of live young after embryo transfer as criteria of sperm survival. Epididymal sperm were frozen in a raffinose-glycerol solution containing either egg yolk (EY) or BSA. Sperm frozen in the presence of EY yielded higher survivals than those frozen in the presence of BSA; when used for IVF, the former resulted in more blastocysts (26% vs. 5%). Comparison between one-step and stepwise dilution of the cryoprotectant revealed no differences in sperm viability as assayed by a vital stain. Nevertheless, IVF using frozen sperm recovered by one-step or stepwise dilution resulted in 33% and 17% blastocyst formation, respectively. When embryos produced with cryopreserved sperm were transferred into recipients, pregnancies (12 of 15 recipients) and the birth of live young (57 pups from 167 embryos transferred) resulted. These results were comparable to those obtained from transferral of embryos produced by IVF with fresh sperm (17 pregnancies from 19 recipients, 87 live young from 197 embryos). Thus, this study has resulted in the development of a simple and efficient cryopreservation procedure for mouse sperm.

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Section: Effect Of Ey On Survival Of Mouse Sperm After Freezing and Tmentioning

confidence: 99%

Birth of Live Mice Resulting from Oocytes Fertilized in Vitro with Cryopreserved Spermatozoa1

Songsasen

Betteridge

1997

Biology of Reproduction

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“…Some lipids are released from sperm membranes during cold shock [20,23]. Egg yolk, a common protectant in cryopreservation media, is crude lipid that interacts with sperm plasma membrane [24][25][26][27], and addition of some lipids to the extender has been suggested to have beneficial effects on cold resistance [28].…”

Section: Introductionmentioning

confidence: 99%

Incorporating Lipids into Boar Sperm Decreases Chilling Sensitivity but Not Capacitation Potential1

Bailey

Buhr

2001

Biology of Reproduction

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Fresh boar sperm were incubated with small unilamellar liposomes composed of either the total lipids extracted from head plasma membranes (HPM) of fresh boar sperm or selected lipids (SL) of five defined phospholipids with specific acyl chains. To optimize fusion, liposomes with 2 mol% octadecyl rhodamine fluorophore in Beltsville Thawing Solution +/- 1 mM CaCl(2) were incubated at 35 degrees C with 1;ts 10(7) or 10(8) spermatozoa/ml and monitored over 60 min, using flow cytometry and fluorescence microscopy. The HPM fused to both sperm concentrations faster than SL but was equivalent by 30 min (10(8) sperm/ml) or 60 min (10(7) sperm/ml; 57.5 +/- 3% and 67.1 +/- 8% sperm fused to HPM and SL, respectively) +/- Ca(2+). Neither HPM nor SL affected onset of capacitation or spontaneous or ionophore-induced acrosome reactions at 0 or 3 h (chlortetracycline and fluorescein isothiocyanate-Pisum sativum agglutinin; n = 3). During cooling and after cryopreservation (n = 4 ejaculates), SL but not HPM significantly improved sperm motility and viability (Sybr14/propidium iodide staining) +/- 20% egg yolk, but egg yolk alone was more effective than SL alone. Liposomes of complex composition can fuse to boar sperm without harming in vitro capacitation or acrosome reaction and reduce sperm chilling sensitivity.

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“…Attempts have been made to characterize the components in egg yolk that provide protection against damage during cooling, freezing and thawing. Phospholipids (Kampschmidt, Mayer & Herman, 1953), phosphatidyl choline (Martin, 1963;Lanz, Pickett & Komarek, 1965;Masuda & Nishikawa, 1972), various lipid extracts and lipoprotein fractions (Bialy, Ludwick, Hess & Ely, 1957;Gebauer, Pickett, Komarek & Gaurya, 1970) and specific lipoproteins (Masuda & Nishikawa, 1972;Foulkes, 1977) have all been reported to provide cryoprotection to spermatozoa.…”

Section: Introductionmentioning

confidence: 99%

A spectrofluorometric investigation, using 1-anilino-naphthalene-8-sulphonate, of the interaction between washed bovine spermatozoa and seminal plasma or egg-yolk lipoprotein

MacDonald¹,

Foulkes²

1981

Reproduction

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Summary. The dye 1-anilino-naphthalene-8-sulphonate (ANS) bound at 94\m=.\5\ m=+-\ 2\m=.\7 pmol per 106 spermatozoa (Kd = 1\m=.\83\ m=+-\0\m=.\13\m=x\ 10\m=-\5 m) and 7\m=.\30\ m=+-\ 0\m=.\17 pmol per mg seminal plasma protein (Kd = 2\m=.\19\ m=+-\0\m=.\06\m=x\10\m=-\5 m). Equilibration of excess ANS with a mixture of washed spermatozoa and seminal plasma resulted in a significant increase in fluorescence over that calculated by summation of the individual levels (P < 0\m=.\05). A less pronounced but significant increase was seen when an egg-yolk lipoprotein, previously shown to have cryoprotective properties, was similarly added to washed spermatozoa (P < 0\m=.\05).This increase in fluorescence was not reduced by washing spermatozoa\p=n-\lipoproteinmixtures, suggesting that an interaction between the lipoprotein and the cell membrane had occurred.

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Motility of Bovine Spermatozoa Extended in “Defined” Diluents

Cited by 25 publications

References 16 publications

Birth of Live Mice Resulting from Oocytes Fertilized in Vitro with Cryopreserved Spermatozoa1

Birth of Live Mice Resulting from Oocytes Fertilized in Vitro with Cryopreserved Spermatozoa1

Incorporating Lipids into Boar Sperm Decreases Chilling Sensitivity but Not Capacitation Potential1

A spectrofluorometric investigation, using 1-anilino-naphthalene-8-sulphonate, of the interaction between washed bovine spermatozoa and seminal plasma or egg-yolk lipoprotein

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