Journal: International Journal of Molecular Sciences

Article Title: Mating Increases CHST10 Activity in Rat Oviductal Mucosa to Induce the Synthesis of HNK-1 Glycoproteins: Possible Role in Sperm–Oviduct Interactions

doi: 10.3390/ijms26073309

Figure Lengend Snippet: Figure 1. Mating increases the carbohydrate sulfotransferase 10 (CHST10) protein level in the oviductal mucosa of rats. Western blot of samples obtained from the oviductal mucosa of unmated (UM1-4) and mated (M1-4) rats. Biological replicates for each group, n = 4. The bar graph represents the intensities of the three CHST10 isoforms detected by the Western blot and normalized to that of tubulin β class I (TUBB). Interquartile range (IQR): Interquartile range. * p < 0.05, indicates significant differences.

Article Snippet: We also evaluated the substrate specificity of human recombinant CHST10 (R&D Systems, cat# 6140-ST) and compared the activity of this enzyme to the sulfotransferase activity detected in oviductal mucosa protein extracts.

Techniques: Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Mating Increases CHST10 Activity in Rat Oviductal Mucosa to Induce the Synthesis of HNK-1 Glycoproteins: Possible Role in Sperm–Oviduct Interactions

doi: 10.3390/ijms26073309

Figure Lengend Snippet: Figure 2. Expression of CHST10 in the rat oviduct. CHST10 was detected via immunofluorescence in oviductal sections obtained from unmated and mated rats. The panels show merged red and blue fluorescence. CHST10: red; and nuclear staining: blue. (First row) ampulla; (second row) isthmus; and (third row) utero-tubal junction (UTJ). The panels in the left column are the negative controls. White arrowheads: immune cells in negative controls; red arrowheads: CHST10 signal resembling the Golgi apparatus location; red arrows: CHST10 in the epithelium cytoplasm. L, Lumen; E, mucosa layer; M, muscular layer. Original magnification, 200× (scale bar, 75 µm). Only the right panels display the image at 630× magnification (scale bar, 25 µm). Images are representative of three independent experiments.

Article Snippet: We also evaluated the substrate specificity of human recombinant CHST10 (R&D Systems, cat# 6140-ST) and compared the activity of this enzyme to the sulfotransferase activity detected in oviductal mucosa protein extracts.

Techniques: Expressing, Immunofluorescence, Fluorescence, Staining

Journal: International Journal of Molecular Sciences

Article Title: Mating Increases CHST10 Activity in Rat Oviductal Mucosa to Induce the Synthesis of HNK-1 Glycoproteins: Possible Role in Sperm–Oviduct Interactions

doi: 10.3390/ijms26073309

Figure Lengend Snippet: Figure 3. Mating increases CHST10 activity in the oviductal mucosa of rats. (A) Michaelis-Menten kinetics of human recombinant CHST10 to evaluate specific acceptor substrates. PGA: phenolph- thalein glucuronic acid; E23G: 17β-estradiol 3-[β-D-glucuronide]. Biological replicates for each group, n = 3. (B) Pattern of acceptor substrate preferences for CHST10 (left) and total protein from the oviductal mucosa of unmated rats (right). CSA: chondroitin sulfate A; E2: 17β-estradiol; E217G: 17β-estradiol 17-[β-D-glucuronide]; DMSO: dimethyl sulfoxide. Biological replicates for each group, n = 3. (C) Plots of sulfotransferase activity measured in the oviductal mucosa of unmated and mated rats using PGA and E23G as acceptor substrates. Biological replicates for each group, n = 8. *** p < 0.001, indicates significant differences.

Article Snippet: We also evaluated the substrate specificity of human recombinant CHST10 (R&D Systems, cat# 6140-ST) and compared the activity of this enzyme to the sulfotransferase activity detected in oviductal mucosa protein extracts.

Techniques: Activity Assay, Recombinant

Journal: International Journal of Molecular Sciences

Article Title: Mating Increases CHST10 Activity in Rat Oviductal Mucosa to Induce the Synthesis of HNK-1 Glycoproteins: Possible Role in Sperm–Oviduct Interactions

doi: 10.3390/ijms26073309

Figure Lengend Snippet: Figure 10. Vaginocervical Stimulation (VCS) increases the level of Chst10 in the oviductal mucosa of rats. The graphs show the RT-qPCR results for Chst10 normalized to the expression of reference genes. The scatter plots represent the normalized individual data points after 2−∆Cq transformation for each individual sample. (A) Oviductal mucosa obtained 3 h after intrauterine injection of 0.1 mL of saline (0.9% w/v NaCl), a suspension of sperm in saline (10 million cells), or diluted seminal vesicle fluid (SVF, 1:1 dilution) into unmated rats. Biological replicates for each group, n = 5. (B) Oviductal mucosa obtained from unmated rats 3 h after VCS or sham stimulation. Biological replicates for each group, n = 5. ** p < 0.01, indicates significant differences.

Article Snippet: We also evaluated the substrate specificity of human recombinant CHST10 (R&D Systems, cat# 6140-ST) and compared the activity of this enzyme to the sulfotransferase activity detected in oviductal mucosa protein extracts.

Techniques: Quantitative RT-PCR, Expressing, Transformation Assay, Injection, Saline, Suspension