A Subpopulation of Morphologically Normal, Motile Spermatozoa Attach to Equine Oviductal Epithelial Cell Monolayers1

Thomas, Philip; Ball, Barry A.; Miller, Paula; Brinsko, Steven P.; Southwood, Louise L.

doi:10.1095/biolreprod51.2.303

Cited by 83 publications

(45 citation statements)

References 42 publications

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“…In contrast to previous results in pigs [11] and cows [52], we found preferential binding of caudal spermatozoa to the isthmus but not ampulla of follicular-phase sows compared to non-cycling gilts. This is consistent with studies in the horse [53] in which the presence of oestrus (but not diestrus) concentrations of steroids in the medium increased the percentage of spermatozoa attaching to both the isthmus and ampulla of the oviduct. These results imply the significant involvement of increased levels of oestrogen in the binding of spermatozoa to oviducts of sexually mature sows compared to pre-pubertal gilts.…”

Section: Discussionsupporting

confidence: 91%

Oviduct binding ability of porcine spermatozoa develops in the epididymis and can be advanced by incubation with caudal fluid

et al. 2015

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Abstract.The sperm reservoir is formed when spermatozoa bind to the epithelium of the utero-tubal junction and caudal isthmus of the oviduct. It is an important mechanism that helps synchronize the meeting of gametes by regulating untimely capacitation and polyspermic fertilisation. This study investigated the influence of epididymal maturation and caudal fluid on the ability of spermatozoa to bind to oviduct epithelium using a model porcine oviduct explant assay. Spermatozoa from the rete testis, middle caput (E2-E3), middle corpus (E6) and cauda (E8) of Large White or Large White x Landrace boars at 10-14 months of age were diluted in modified Androhep solution and incubated with porcine oviduct explants. Results reported in this study support our hypothesis that testicular spermatozoa need to pass through the regions of the epididymis in order to acquire the ability to bind to the oviduct. There was a sequential increase in the number of spermatozoa that bound to oviduct explants from the rete testis to caudal epididymis. Binding of caudal spermatozoa to isthmic explants was the highest (15.0 ± 1.2 spermatozoa per 1.25 mm 2 ; mean ± standard error of the mean; P ≤ 0.05) and lowest by spermatozoa from the rete testis (2.0 ± 0.3 per 1.25 mm 2 ), and higher to isthmus from sows compared to gilts (35.8 ± 6.7 per 1.25 mm 2 vs. 14.8 ± 3.0 per 1.25 mm 2 ; P ≤ 0.05). Binding of ejaculated spermatozoa to porcine isthmus was higher than for caudal spermatozoa (26.3 ± 1.4 per 1.25 mm 2 vs. 15.0 ± 0.8 per 1.25 mm 2 ; P ≤ 0.05), and higher to porcine than to bovine isthmus (26.3 ± 2.3 per 1.25 mm 2 vs. 18.8 ± 1.9 per 1.25 mm 2 ; P ≤ 0.05). Incubation of spermatozoa from the caput and corpus in caudal fluid 2 increased the ability of spermatozoa to bind to oviduct epithelium (P ≤ 0.05). In conclusion, the capacity of testicular spermatozoa to bind to oviduct epithelium increases during their maturation in the epididymis, and can be advanced by components of the caudal fluid.

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Section: Discussionsupporting

confidence: 91%

Oviduct binding ability of porcine spermatozoa develops in the epididymis and can be advanced by incubation with caudal fluid

et al. 2015

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“…However, monolayers as an experimental model are advantageous in that they provide a ready source of oviductal material that is produced by in vitro culture. In addition, as an adherent sheet of cells, binding of spermatozoa to oviductal monolayers can be assessed by direct (Raychoudhury and Suarez, 1991;Gualtieri and Talevi, 2000) or indirect counting methods (by subtracting the number of spermatozoa recovered after co-culture) (Thomas et al, 1994;Chian and Sirard, 1995).…”

Section: Discussionmentioning

confidence: 99%

Carbohydrate mediation of boar sperm binding to oviductal epithelial cells in vitro

Bredl²,

Holt³

et al. 2001

Reproduction

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ResearchAfter mating, mammalian spermatozoa are transported to the lower oviductal isthmus. Spermatozoa are sequestered at the isthmus by attaching and interacting with oviductal epithelial cells, hence forming a sperm reservoir. In several mammalian species, specific carbohydrates mediate spermoviductal epithelial cell binding. A quantitative in vitro free cell bioassay was developed to investigate the involvement of carbohydrate recognition in pig spermoviductal epithelial cell interactions. This assay was validated. The sensitivity of the assay was such that it was possible to discriminate between different sperm concentrations and sperm-oviductal epithelial cell co-incubation periods, spermatozoa with damaged plasma membranes and epithelial cells of non-reproductive origin. Optimal conditions were used to incubate spermatozoa and oviductal epithelial cells in the presence of six hexose sugars at concentrations of 0, 2, 10 and 50 mmol l -1 . A significant (P р 0.05) reduction in the binding of spermatozoa to the oviductal epithelium was detected with 2, 10 and 50 mmol maltose l -1 , 50 mmol lactose l -1 and 50 mmol mannose l -1 . These findings support the hypothesis that attachment of pig spermatozoa to oviductal epithelium before fertilization is mediated by carbohydrate recognition.

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“…From various studies undertaken in different species it has been hypothesized that the binding of spermatozoa to the oviduct presents a mechanism for selecting a competent sperm population. Thomas et al (1994) and Petrunkina et al (2001) have reported that in equines and pigs, binding capacity depends on sperm quality, for example, morphology. Furthermore, spermatozoa bound to oviductal epithelium in vitro have intact acrosomes (Suarez et al, 1991;Thomas and Ball, 1996), thereby indicating a selective function of sperm-oviduct binding.…”

Section: Discussionmentioning

confidence: 99%

Kinetic characterization of the changes in protein tyrosine phosphorylation of membranes, cytosolic Ca2+ concentration and viability in boar sperm populations selected by binding to oviductal epithelial cells

Friedrich²,

Drommer³

et al. 2001

Reproduction

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On reaching the oviduct, spermatozoa are retained in the isthmic region of the oviduct until ovulation occurs. The essential steps of capacitation are co-ordinated in this region. In this study, a primary cell culture system of oviductal epithelial cells was established to investigate sperm binding to oviductal epithelium and modulation of sperm function during incubation under capacitating conditions in co-culture with oviductal epithelial cells. Epithelial cells were stripped from the oviducts of sows and cultivated for 5-7 days on Lab-Tek Chamber slides on Matrigel. The preparations on chamber slides and suspensions of control spermatozoa were incubated for 3 h in Tyrode's albumin lactate pyruvate (TALP) medium. At 3, 30, 60, 90 and 180 min the free-swimming spermatozoa were collected by washing, and membrane integrity, tyrosine phosphorylation patterns and [Ca(2+)](i) of bound, unbound and control spermatozoa were assessed with fluorescent probes (propidium iodide, Cy-3 and fluo-3-AM). The cells bound to oviductal epithelial cells showed reduced cytosolic Ca(2+) concentration, reduced and almost absent tyrosine phosphorylation of membrane proteins and higher viability at the time of the first sampling. Increases in Ca(2+) concentration and cell death occurred much more slowly during incubation in cells bound to oviductal epithelial cells compared with free-swimming spermatozoa, and no changes in tyrosine phosphorylation were observed. The preferential binding of viable, low-Ca(2+) cells with suppressed tyrosine phosphorylation and slower functional modulation of boar spermatozoa attached to oviductal epithelial cells might represent a mechanism for selecting functionally competent spermatozoa and prolonging their lifespan by delaying capacitation in the oviductal reservoir.

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A Subpopulation of Morphologically Normal, Motile Spermatozoa Attach to Equine Oviductal Epithelial Cell Monolayers1

Cited by 83 publications

References 42 publications

Oviduct binding ability of porcine spermatozoa develops in the epididymis and can be advanced by incubation with caudal fluid

Oviduct binding ability of porcine spermatozoa develops in the epididymis and can be advanced by incubation with caudal fluid

Carbohydrate mediation of boar sperm binding to oviductal epithelial cells in vitro

Kinetic characterization of the changes in protein tyrosine phosphorylation of membranes, cytosolic Ca2+ concentration and viability in boar sperm populations selected by binding to oviductal epithelial cells

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