Andrology: The interaction in vitro of human spermatozoa with epithelial cells from the human uterine (Fallopian) tube

Pacey, Allan; Hill, Chris; Scudamore, I. W.; Warren, M.A.; Barratt, Christopher L.R.; Cooke, I. D.

doi:10.1093/oxfordjournals.humrep.a135943

Cited by 88 publications

(39 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The majority of boar spermatozoa (63.1%) bound FITC-labelled sAPM proteins at the apical region of the sperm head; this is the region that associates with OEC in vivo (Rodriguez-Martinez 2007) and suggests that the sAPM-sperm interaction mimics the natural situation. This also correlates with binding patterns and interactions seen in other studies (Pollard et al 1991, Pacey et al 1995. Binding over the anterior acrosome also coincides with the distribution of sulphated glycolipids reported by Gadella et al (1995); interestingly, the sulphated glycolipids are known to bind specifically to members of the HSP70 family, especially HSPA8, through specific binding sites on the protein molecule (Boulanger et al 1995.…”

Section: Discussionsupporting

confidence: 89%

“…In general, mammals are only able to store spermatozoa for a few days in the female reproductive tract (Rodriguez-Martinez et al 2005, Rodriguez-Martinez 2007, although some bat species have evolved successful means of storing spermatozoa for several months over winter in the uterus or oviduct (Racey & Potts 1970, Racey 1979, Racey et al 1987. The mechanisms involved in prolonging sperm survival in the female reproductive tract have remained largely elusive, although the widely observed (Hunter 1981, Pacey et al 1995, Hunter & Rodriguez-Martinez 2004 formation of intimate associations between spermatozoa and the oviductal epithelial surface suggests that membrane-membrane contact is likely to be important.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Effects of HSPA8, an evolutionarily conserved oviductal protein, on boar and bull spermatozoa

Elliott¹,

Lloyd²,

Fazeli³

et al. 2009

Reproduction

View full text Add to dashboard Cite

Previous studies have shown that a soluble protein fraction derived from preparations of apical plasma membrane (APM) of the oviductal epithelium enhances the in vitro survival of mammalian spermatozoa. Here, we show that the survival enhancing property of the soluble protein fraction seems to depend significantly upon heat shock 70 kDa protein 8 (HSPA8 previously known as HSPA10). The following findings in the present study enabled us to draw this conclusion: first, using proteomic analysis, we identified a subset of 70 kDa oviductal surface proteins that bound to spermatozoa, one of which was HSPA8. Second, pre-treatment of the soluble protein fraction with anti-HSPA8 antibody reduced the 24 h (at 39 8C) sperm survival enhancement effect normally induced by the presence of 200 mg/ml soluble APM proteins. Third, complementary experiments showed that substituting the soluble protein fraction with bovine recombinant HSPA8 (0.5-2 mg/ml) also elicited the sperm survival effect. Finally, we also tested the effect of bovine recombinant HSPA8 on bull spermatozoa and found similar, dose-responsive, sperm survival promoting effects. The conserved nature of HSPA8 between mammalian species suggests that this protein may represent a common biological mechanism for the maintenance of sperm survival in the oviduct.

show abstract

Section: Discussionsupporting

confidence: 89%

Section: Introductionmentioning

confidence: 99%

Effects of HSPA8, an evolutionarily conserved oviductal protein, on boar and bull spermatozoa

Elliott¹,

Lloyd²,

Fazeli³

et al. 2009

Reproduction

View full text Add to dashboard Cite

show abstract

“…Human endosalpingeal cells were obtained from four women who donated their fallopian tubes for research purposes at the time of their hysterectomy. Details of the surgical procedure and methods used to provide primary human endosalpingeal cells for culture and experimentation are described by Pacey et al (31). All women were previously fertile and had normal endocrinology.…”

Section: Methodsmentioning

confidence: 99%

“…maintain their polarity, actively beating cilia on their apical surface suggest that in their unpassaged state they still have retained some polarity. Moreover, they are considered to be a good model of the endosalpinx in other experimental systems (20,28,31). The decision to use both serovars E and LGV in this experiment was made on the basis of known differences between them, in terms of their response to GAGs (5,6,12) and the differences in the types of clinical presentations that result from their infection (12,29).…”

Section: Fig 3 Elution Profiles Of Radiolabeled Gags Prepared From mentioning

confidence: 99%

Infectivity ofChlamydia trachomatisSerovar LGV but Not E Is Dependent on Host Cell Heparan Sulfate

et al. 2001

View full text Add to dashboard Cite

The ability of heparan sulfate, heparin, and other glycosaminoglycans to inhibit the infectivity of Chlamydia trachomatis serovars E and LGV was examined using a simple competitive inhibition assay with three cell types from the human female reproductive tract, including primary human endosalpingeal cells. With the majority of the glycosaminoglycans tested, LGV was more significantly inhibited than serovar E. We have compared chlamydial infectivity between a wild-type Chinese hamster ovary cell line and two glycosaminoglycan-deficient cell lines.LGV was shown to be unable to infect heparan sulfate-deficient and GAG-deficient Chinese hamster ovary cell lines, whereas the E serovar infected these cells as efficiently as the control (nondeficient) cells. These two sets of experiments confirmed that serovar LGV is more dependent on a heparan sulfate-related mechanism of infectivity than is serovar E. This is further supported by the fact that attempts to purify a heparan sulfate-like molecule from either serovar cultured in glycosaminoglycan-deficient cell lines were nonproductive. Previous reports have suggested that chlamydia are able to produce a heparan sulfate-like molecule that is important for attachment and infectivity. We have attempted to detect possible binding of a specific heparan sulfate antibody to C. trachomatis by flow cytometry. Results showed no binding of the heparan sulfate antibody to C. trachomatis serovar LGV or E. Our results strongly indicate that chlamydiae do not produce a heparan sulfate-like molecule but rather use host cell heparan sulfate in order to infect cells.

show abstract

“…In mammalian species, the isthmus of the oviduct serves as a temporary sperm reservoir before ovulation with spermatozoa bound to the ciliated oviductal epithelial cells through a carbohydrate-mediated event (Suarez and Pacey, 2006;Topfer-Petersen, 1999). Although definitive evidence for a distinct sperm reservoir in human Fallopian tubes is lacking, human spermatozoa attached to the endosalpingeal epithelium intermittently in vitro (Pacey et al, 1995). Our unpublished data also show that purified sperm FUT binds to the ciliated oviductal epithelial cells.…”

Section: Journal Of Cell Sciencementioning

confidence: 56%

Glycodelin-A interacts with fucosyltransferase on human sperm plasma membrane to inhibit spermatozoa-zona pellucida binding

Chiu

Chung

Koistinen

et al. 2007

Journal of Cell Science

View full text Add to dashboard Cite

Journal of Cell Science 34 suggest that the glycodelin receptor(s) and zona pellucida glycoprotein receptor(s) are closely related. The objectives of this study were to identify the receptor of glycodelin-A in human spermatozoa and to characterize its interaction with human zona pellucida. Results Fucosyltransferase 5 (FUT5) is a sperm surface glycodelin-A binding proteinThe results of identification of glycodelin-A-bound sperm surface protein(s) are shown in Figs 1, 2. Sulfosuccinimidyl-2-[6-(biotinamido)-2-(p-azidobenzamido)-hexanoamido]ethyl-1,3Ј-dithioproionate (Sulfo-SBED)-conjugated glycodelin-A was covalently bound to spermatozoa after photoactivation.Treatment of the glycodelin-A-sperm surface protein complexes with dithiothreitol (DTT; reducing conditions) resulted in reduction of the azo linkage and transfer of the biotin label from glycodelin-A to the sperm proteins. Thus, biotinylated glycodelin-A (Fig. 1A,B, lane 1) showed a single biotin band with molecular size ~30 kDa under non-reducing conditions, which was lost after DTT treatment (Fig. 1A,B, lane 2). One major biotinylated complex with size ~93 kDa appeared after incubation of glycodelin-A (Fig. 1A,B, lane 3) with the sperm extract. Treatment with DTT revealed a major biotinlabeled complex of size~65 kDa (Fig. 1A,B, lane 4). Some minor bands of smaller sizes were also seen. By contrast, glycodelin-A did not interact with fibroblasts and nonspecifically bound glycodelin-A was removed by washing of the cells after incubation. This is evidenced by the absence of a band in sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE; Fig. 1A,B, lane 6). Deglycosylated glycodelin-A did not interact with human spermatozoa (data not shown).Mass-spectrometry (MS) analysis of the tryptic digests and a database search identified the ~65 kDa glycodelin-A bound protein as FUT5 (Table 1). The six peptide sequences showed >90% match to the MS spectra and confirmed the protein identity at high confidence level. Two of the identified peptides, SFSWALAFCK and TELVAWAVSNWKPDSAR, were unique to FUT5 among fucosyltransferase 3 (FUT3), FUT5 and fucosyltransferase 6 (FUT6). The identity of the protein was confirmed by western blot analysis using the anti-FUT5 antibody, N-18 (Fig. 1C). The immunoreactivity was associated with bands of sizes ~93 and ~65 kDa, corresponding to the glycodelin-A-FUT complex in non-reducing conditions (Fig. 1C, lane 3) and FUT in reducing conditions (Fig. 1C, lane 4), respectively. Presence of FUT protein in human spermatozoaRecombinant FUT proteins were expressed in inclusion bodies and constituted for approximately 50% of the total protein of the transformed Escherichia coli. The yield of soluble protein was approximately 30-66 mg/l of culture. The expected sizes of recombinant FUT3, FUT5 and FUT6 are 51, 58 and 61 kDa, respectively. These recombinant FUTs were used to determine the specificities of the anti-FUT antibodies against different regions of the enzymes.Owing to the high sequence similarity (~90%) at the Cterminals, th...

show abstract

Andrology: The interaction in vitro of human spermatozoa with epithelial cells from the human uterine (Fallopian) tube

Cited by 88 publications

References 15 publications

Effects of HSPA8, an evolutionarily conserved oviductal protein, on boar and bull spermatozoa

Effects of HSPA8, an evolutionarily conserved oviductal protein, on boar and bull spermatozoa

Infectivity ofChlamydia trachomatisSerovar LGV but Not E Is Dependent on Host Cell Heparan Sulfate

Glycodelin-A interacts with fucosyltransferase on human sperm plasma membrane to inhibit spermatozoa-zona pellucida binding

Contact Info

Product

Resources

About