Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: 'ZP domain' of human zona pellucida glycoprotein-1 binds to human spermatozoa and induces acrosomal exocytosis

doi: 10.1186/1477-7827-8-110

Figure Lengend Snippet: Profile of lectins binding to baculovirus-expressed human ZP1 273-551aa in ELISA . Microtitration plates were coated with the baculovirus-expressed recombinant ZP1 273-551aa (open bar; 500 ng/well) and processed for evaluation of binding to 21 different biotinylated lectins in an ELISA as described in Methods . As an internal control, microtitration plates coated with same amount of E. coli -expressed recombinant human ZP1 273-551aa (grey bar; unpublished observations) were also used. Values are expressed as absorbance obtained with various lectins binding to the respective recombinant protein, after deducting the non-specific binding of the lectins to the uncoated wells. Each bar represents a mean of duplicate experiments and standard deviation of the absorbance values. The lectins tested were GSL-I: Griffon simplicifolia lectin I, SBA: Soybean agglutinin, DSL: Datura stramonium lectin, PSA: Pisum sativum agglutinin, ConA: Concanavalin A, ECL: Erythrina cristagalli lectin, LCA: Lens culinaris agglutinin, DBA: Dolichos biflorus agglutinin, LEL: Lycopersicon esculentum lectin, PHA-L: Phaseolus vulgaris leucoagglutinin, UEA-I: Ulex europaeus agglutinin I, STL: Solanum tuberosum lectin, PHA-E: Phaseolus vulgaris erythroagglutinin, RCA: Ricinus communis agglutinin, VVA: Vicia villosa agglutinin, SJA: Sophora japonica agglutinin, PNA: Peanut agglutinin, S- WGA: Succinylated Wheat germ agglutinin, Jacalin, WGA: Wheat Germ agglutinin and GSL II: Griffonia simplicifolia lectin II.

Article Snippet: Post-incubation, sperm were processed for simultaneous assessment of the status of acrosome by double labelling with 5 μg/ml tetramethyl rhodamine isothiocyanate conjugated Pisum sativum agglutinin (TRITC-PSA; Vector Laboratories Inc., Burlingame, CA, USA) and for binding of FITC-labelled human ZP1 273-551aa as described earlier [ ].

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Recombinant, Standard Deviation

Journal: PLoS ONE

Article Title: Human Herpesvirus-6A/B Binds to Spermatozoa Acrosome and Is the Most Prevalent Herpesvirus in Semen from Sperm Donors

doi: 10.1371/journal.pone.0048810

Figure Lengend Snippet: Semen samples were pre-incubated with HHV-6B for one hr followed by incubation with either vehicle or calcium ionophore for 30 min to induce an acrosome reaction (AR). After incubation with both calcium ionophore and virus the samples were washed twice in PBS and stained with anti-gp60/110 or anti-IgG 2b followed by a goat-anti mouse (GaM)-FITC concomitantly with rhodamine-conjugated pisum sativum (PSA). The PSA (red) stains intact acrosomes and leaves acrosome reacted sperm unstained. (A-E) Different combinations of pre-treatment and subsequent staining are indicated at the bottom of the figure.

Article Snippet: Samples were washed, stained and incubated with 1% paraformaldehyde as described above, only this time with co-staining using rhodamine-conjugated pisum sativum (PSA) (Vector Laboratories, Burlingame, CA, USA).

Techniques: Incubation, Staining