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Journal ArticleDOI

Effect of Sperm Cryopreservation and Supplementing Semen Doses with Seminal Plasma on the Establishment of a Sperm Reservoir in Gilts

01 Apr 2007-Reproduction in Domestic Animals (Reprod Domest Anim)-Vol. 42, Iss: 2, pp 149-152
TL;DR: It is demonstrated that the size of the sperm reservoir is markedly reduced in gilts inseminated with FT sperm, suggesting that either it did not reverse cryocapacitation or that such a reversal does not impact the in vivo ability to create a sperm reservoir.
Abstract: Frozen-thawed (FT) boar sperm have a reduced fertile life, due in part to a capacitation-like status induced by cooling. Reversal of this cryocapacitation in vitro by exposure to boar seminal plasma (SP) has been demonstrated. The objective of these studies was to determine the effect of SP on the ability of FT sperm to create an oviductal sperm reservoir following artificial insemination (AI). In Experiment one, 35 pre-pubertal gilts were injected (IM) with 400 IU eCG plus 200 IU hCG to induce oestrus. At detection of oestrus, gilts were inseminated with 3 x 10(9) live sperm, either fresh (FS; n = 13), FT (n = 10), or FT supplemented with 10% v/v SP (n = 12). Gilts were killed 8 h later, their reproductive tracts recovered and the uterotubal junctions (UTJs) flushed to recover sperm. Fewer (p < 0.01) sperm were recovered following FT, compared to FS, inseminations, and there was no evident effect of SP. In Experiment two, 30 pre-pubertal gilts received IM injections of 1000 IU eCG followed by 5 mg pLH 80 h later to control time of ovulation. Gilts were inseminated with 3 x 10(9) live FS sperm (n = 6), FT sperm (n = 15) or FT sperm plus 10% SP (n = 9) at 12 h before ovulation and then sacrificed 8 h later. The UTJs were dissected and flushed for sperm recovery. Fewest (p < 0.001) sperm were recovered following FT insemination and there was no evident effect of SP. These data demonstrate that the size of the sperm reservoir is markedly reduced in gilts inseminated with FT sperm. However, the lack of effect of SP suggested that either it did not reverse cryocapacitation or that such a reversal does not impact the in vivo ability to create a sperm reservoir.
Citations
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Journal ArticleDOI
TL;DR: In this review, different types of membrane stress are considered, as well as novel surface manipulation methods to improve sperm stability.
Abstract: Spermatozoa interact with their immediate environment and this contact remodels the sperm surface in preparation for fertilisation. These fundamental membrane changes will be critically covered in this review with special emphasis on the very specific surface destabilisation event, capacitation. This process involves very subtle and intricate modifications of the sperm membrane including removal of suppression (decapacitation) factors and changes in the lateral organisation of the proteins and lipids of the sperm surface. Processing of sperm for assisted reproduction (storage, sex-sorting, etc.) subjects spermatozoa to numerous stressors, and it is possible that this processing overrides such delicate processes resulting in sperm instability and cell damage. To improve sperm quality, novel mechanisms must be used to stabilise the sperm surface during handling. In this review, different types of membrane stress are considered, as well as novel surface manipulation methods to improve sperm stability.

168 citations


Cites background from "Effect of Sperm Cryopreservation an..."

  • ...While other studies have found no effects on fertility (Abad et al. 2007a) or on the establishment of an oviductal sperm reservoir (Abad et al. 2007b)....

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Journal ArticleDOI
TL;DR: The addition of SP proteins to spermatozoa before and/or after cooling is able to minimize cryoinjury effects and the major proteins in ram SP which are able to protect and repair the cold-shock damage to sperm contain fibronectin-II domains.
Abstract: The role of seminal plasma (SP) in mammalian sperm function remains largely a matter of speculation as both inhibitory and stimulating effects have been found. Specific components of SP, particularly proteins, are adsorbed onto the surface of ejaculated sperm as they pass through the male and female reproductive tracts. These sperm coating components seem to have the important function of maintaining the stability of the membrane up to the process of capacitation (decapacitation factors). Therefore, they must be removed, modified or masked before the spermatozoa undergo the acrosome reaction, an essential process for successful fertilization. It is well known that low temperatures alter the function of spermatozoa. Cold shock results in the destabilization of sperm membranes and impairment of sperm function, and it is also well known that ram spermatozoa are more sensitive to cold-shock stress than those of other species. The addition of SP proteins to spermatozoa before and/or after cooling is able to minimize cryoinjury effects. The major proteins in ram SP which are able to protect and repair the cold-shock damage to sperm contain fibronectin-II domains. The significance of this domain and the role of these proteins in sperm capacitation and gamete interaction are discussed.

150 citations


Cites background or result from "Effect of Sperm Cryopreservation an..."

  • ...Conversely, the addition of SP proteins did not affect fertility of boar (Abad et al. 2007) or ram spermatozoa (El-Hajj Ghaoui et al. 2007; O’Meara et al. 2007)....

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  • ...Conversely, the addition of SP proteins did not affect fertility of boar (Abad et al. 2007) or ram spermatozoa (El-Hajj Ghaoui et al....

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  • ...2007), and that other aspects of cryoinjury in addition to cryocapacitation may be involved on the establishment of a sperm reservoir (Abad et al. 2007)....

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  • ...…and recent results suggest that controlled cooling during boar semen cryopreservation did not induce this premature capacitation (Saravia et al. 2007), and that other aspects of cryoinjury in addition to cryocapacitation may be involved on the establishment of a sperm reservoir (Abad et al. 2007)....

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Journal ArticleDOI
TL;DR: In sum, artificial insemination with frozen‐thawed boar sperm can provide reasonable fertility outcomes, if freezable ejaculates, specific additives, and appropriate Insemination techniques are used.
Abstract: Artificial insemination with frozen-thawed semen in pigs is not a routine technique; its use is restricted to specific cases, such as preservation of valuable genetic material (germplasm banks), safety strategies in case of natural disasters, long-distance transport of sperm, and in combination with sex-sorting. Cryoinjuries resulting from freeze-thawing protocols are a major concern with regard to the fertilization capacity of the treated sperm, which is lower than that of liquid-stored semen. Here, we provide an overview of artificial insemination using cryopreserved sperm, and summarize the factors that influence cryopreservation success before, during, and after freeze-thaw (i.e., sperm selection before starting the cryopreservation process, holding time, use of cryoprotectants, and rates of freezing and thawing) and that are driving the identification of biomarkers to predict sensitivity to cryodamage. Three different artificial insemination techniques (conventional or intracervical; intrauterine; and deep intrauterine) are also discussed with regards to their relevance when using frozen-thawed semen. Finally, we review the use of additives to freezing and thawing media, given reports that they may maintain and improve the quality and fertilizing capacity of frozen-thawed sperm. In sum, artificial insemination with frozen-thawed boar sperm can provide reasonable fertility outcomes, if freezable ejaculates, specific additives, and appropriate insemination techniques are used.

80 citations

Journal ArticleDOI
TL;DR: Differential effects on motility were not clearly accompanied by biologically related modifications of sperm membrane or chromatin structure and it was presumably related to different concentrations or either SP proteins or bicarbonate in the different ejaculate portions.

74 citations


Cites methods from "Effect of Sperm Cryopreservation an..."

  • ...Traditionally, boar spermatozoa have been exposed to various proportions (between 5 and 100% v/v) of whole SP during handling for AI [5,10–13], cooling [14,15] and cryopreservation [9,16] or post-thaw (PT) [17,18], for in vitro fertilization (IVF) [19] or even after procedures such as flow cytometric sorting for chromosomal sex [20–22]....

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Journal ArticleDOI
TL;DR: An adverse effect of inseminating FT sperm on sperm quality and sow fertility is confirmed but it is suggested that thawing FT sperm in 50% SP may partially alleviate these adverse effects.

65 citations


Cites background or result from "Effect of Sperm Cryopreservation an..."

  • ...…et al., 2008), associated with an ncrease in DNA fragmentation (Fraser and Strzezek, 2007). apacitated spermatozoa do not form a functional sperm eservoir (Fazeli et al., 1999; Tienthai et al., 2004) and, simlarly, cryopreserved sperm do not form a normal sperm eservoir (Abad et al., 2007a)....

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  • ...In contrast to our previous study where we thawed FT sperm in 10% seminal plasma (Abad et al., 2007b), the present study suggests a beneficial effect of thawing FT sperm in 50% seminal plasma on subsequent litter size....

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  • ...In contrast, we previously determined that extending thawed semen in 10% seminal plasma had no effect sperm’s ability to form a functional sperm reservoir in the oviducts (Abad et al., 2007a) nor on sow fertility if inseminated 2 h or 12 h before predicted time of ovulation (Abad et al., 2007b)....

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  • ...However, when boar semen was supplemented ith 0% or 10% seminal plasma and inseminated within 2 h r 12 h of the predicted time of ovulation, there was no ffect of timing of insemination or seminal plasma on sow ertility (Abad et al., 2007b)....

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References
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Journal ArticleDOI
TL;DR: The problems, aspects and methods of liquid storage and freeze-thawing of boar semen are discussed and a review is given on examination of spermatozoa by the recent fluorescent staining methods.

718 citations

Journal ArticleDOI
TL;DR: The lack of massive sperm capacitation in the SR and the diverse individual response to capacitation shown by tubal spermatozoa would relate both to the insurance of full sperm viability before ovulation and the presence of spermatozosa at different stages of capacitations in the upper oviduct, thus maximizing the chances of normal fertilization.

228 citations

Journal ArticleDOI
TL;DR: The approach was to vary the proportions of uncapacitated, capacitated, or acrosome-reacted boar spermatozoa in suspension using long preincubation and lysophosphatidylcholine treatment of semen prior to a very short incubation with OEC or LLC-PK1 cells.
Abstract: After mating, inseminated spermatozoa are transported to the oviduct They attach to and interact with oviductal epithelial cells (OEC) To investigate sperm-OEC interactions, we used chlortetracycline to study the capacitation status of boar spermatozoa in coculture with homologous OEC and cells of nonreproductive origin (LLC-PK1, porcine kidney epithelial cell line) Boar spermatozoa were cocultured with OEC and LLC-PK1 cells for 15, 60, 120, or 240 min The proportion of capacitated spermatozoa in coculture with the isthmic and ampullar cells increased significantly (p < 005) during incubation However, most spermatozoa in coculture with LLC-PK1 cells or blank (medium only) remained uncapacitated In addition, preferential binding of uncapacitated, capacitated, or acrosome-reacted boar spermatozoa to OEC and the other cell type was investigated Our approach was to vary the proportions of uncapacitated, capacitated, or acrosome-reacted boar spermatozoa in suspension using long preincubation and lysophosphatidylcholine treatment of semen prior to a very short incubation with OEC or LLC-PK1 cells The results showed that the majority of spermatozoa that were bound to OEC or LLC-PK1 cells were uncapacitated and that a significant relationship existed between the relative proportion of uncapacitated spermatozoa in the control samples and those bound to LLC-PK1 cells (r2 = 043, p < 0005) However, there was no correlation between the proportion of uncapacitated spermatozoa in the control samples and the proportion of those bound to isthmic or ampullar cells In conclusion, the results clearly demonstrated the specific nature of the sperm-OEC interaction in the porcine species This interaction is initiated by uncapacitated spermatozoa binding to OEC and is continued by the induction of capacitation in cocultured spermatozoa

220 citations

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These data demonstrate that the size of the sperm reservoir is markedly reduced in gilts inseminated with FT sperm.