Limits...
Fibroblast Growth Factor Receptors (FGFRs) in Human Sperm: Expression, Functionality and Involvement in Motility Regulation.

Saucedo L, Buffa GN, Rosso M, Guillardoy T, Góngora A, Munuce MJ, Vazquez-Levin MH, Marín-Briggiler C - PLoS ONE (2015)

Bottom Line: Fibroblast growth factors receptors (FGFRs) have been widely characterized in somatic cells, but there is scarce evidence of their expression and function in mammalian gametes.Incubation with FGF2 led to a significant increase in the percentage of total and progressive sperm motility, as well as in sperm kinematics.In addition to confirming the expression of FGFRs in germ cells of the human testis, our study describes for the first time the presence, localization and functionality of human sperm FGFRs, and provides evidence of the beneficial effect of FGF2 upon sperm motility.

View Article: PubMed Central - PubMed

Affiliation: Instituto de Biología y Medicina Experimental (IBYME), CONICET-FIBYME, Buenos Aires, Argentina.

ABSTRACT
Fibroblast growth factors receptors (FGFRs) have been widely characterized in somatic cells, but there is scarce evidence of their expression and function in mammalian gametes. The objective of the present study was to evaluate the expression of FGFRs in human male germ cells, to determine sperm FGFR activation by the FGF2 ligand and their participation in the regulation of sperm motility. The expression of FGFR1, 2, 3 and 4 mRNAs and proteins in human testis and localization of these receptors in germ cells of the seminiferous epithelium was demonstrated. In ejaculated sperm, FGFRs were localized to the acrosomal region and flagellum. Sperm exposure to FGF2 caused an increase in flagellar FGFR phosphorylation and activation of extracellular signal-regulated kinase (ERK) and protein kinase B (PKB or Akt) signaling pathways. Incubation with FGF2 led to a significant increase in the percentage of total and progressive sperm motility, as well as in sperm kinematics. All responses were prevented by sperm preincubation with BGJ398, a specific inhibitor of FGFR tyrosine kinase activity. In addition to confirming the expression of FGFRs in germ cells of the human testis, our study describes for the first time the presence, localization and functionality of human sperm FGFRs, and provides evidence of the beneficial effect of FGF2 upon sperm motility.

No MeSH data available.


Related in: MedlinePlus

Localization of FGFRs in human sperm.Sperm cells were stained with anti FGFR1, FGFR2, FGFR3 and FGFR4 or rabbit IgG and a secondary antibody labeled with Cy3. The corresponding fields stained with FITC-PSA to assess acrosomal status are shown. Bar: 10 μm. On the right, a representative image of individual sperm is depicted; (A) sperm stained with anti FGFR antibody and Cy3-conjugated secondary antibody, (B) FITC-PSA, (C) merge.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4430232&req=5

pone.0127297.g003: Localization of FGFRs in human sperm.Sperm cells were stained with anti FGFR1, FGFR2, FGFR3 and FGFR4 or rabbit IgG and a secondary antibody labeled with Cy3. The corresponding fields stained with FITC-PSA to assess acrosomal status are shown. Bar: 10 μm. On the right, a representative image of individual sperm is depicted; (A) sperm stained with anti FGFR antibody and Cy3-conjugated secondary antibody, (B) FITC-PSA, (C) merge.

Mentions: To determine FGFR localization in human sperm, immunofluorescence analysis with anti FGFR antibodies was carried out (Fig 3). In all sperm suspensions evaluated, over 90% of the cells were classified as acrosome intact by means of the FITC-PSA staining. FGFR1 was localized to the principal piece of the flagellum in 100% of the cells and to the acrosomal region in 81 ± 2% of the sperm (n = 8). FGFR2 was detected along the flagellum of all cells and in the acrosomal region in 84 ± 1% of the cells (n = 7); in addition, a strong signal in the neck was found in some cells. Sperm stained with anti FGFR3 showed label in the midpiece of the flagellum, with a faint signal in the principal piece, and in the acrosomal region (80 ± 4%) (n = 8). All cells stained with anti FGFR4 depicted an intense signal along the flagellum and 69 ± 10% of the sperm showed label in the acrosomal region (n = 5). FGFR immunodetection was specific, since no staining was observed when primary antibodies were replaced by rabbit IgG (Fig 3) or when the antibodies were blocked with the respective peptides (S3 Fig).


Fibroblast Growth Factor Receptors (FGFRs) in Human Sperm: Expression, Functionality and Involvement in Motility Regulation.

Saucedo L, Buffa GN, Rosso M, Guillardoy T, Góngora A, Munuce MJ, Vazquez-Levin MH, Marín-Briggiler C - PLoS ONE (2015)

Localization of FGFRs in human sperm.Sperm cells were stained with anti FGFR1, FGFR2, FGFR3 and FGFR4 or rabbit IgG and a secondary antibody labeled with Cy3. The corresponding fields stained with FITC-PSA to assess acrosomal status are shown. Bar: 10 μm. On the right, a representative image of individual sperm is depicted; (A) sperm stained with anti FGFR antibody and Cy3-conjugated secondary antibody, (B) FITC-PSA, (C) merge.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4430232&req=5

pone.0127297.g003: Localization of FGFRs in human sperm.Sperm cells were stained with anti FGFR1, FGFR2, FGFR3 and FGFR4 or rabbit IgG and a secondary antibody labeled with Cy3. The corresponding fields stained with FITC-PSA to assess acrosomal status are shown. Bar: 10 μm. On the right, a representative image of individual sperm is depicted; (A) sperm stained with anti FGFR antibody and Cy3-conjugated secondary antibody, (B) FITC-PSA, (C) merge.
Mentions: To determine FGFR localization in human sperm, immunofluorescence analysis with anti FGFR antibodies was carried out (Fig 3). In all sperm suspensions evaluated, over 90% of the cells were classified as acrosome intact by means of the FITC-PSA staining. FGFR1 was localized to the principal piece of the flagellum in 100% of the cells and to the acrosomal region in 81 ± 2% of the sperm (n = 8). FGFR2 was detected along the flagellum of all cells and in the acrosomal region in 84 ± 1% of the cells (n = 7); in addition, a strong signal in the neck was found in some cells. Sperm stained with anti FGFR3 showed label in the midpiece of the flagellum, with a faint signal in the principal piece, and in the acrosomal region (80 ± 4%) (n = 8). All cells stained with anti FGFR4 depicted an intense signal along the flagellum and 69 ± 10% of the sperm showed label in the acrosomal region (n = 5). FGFR immunodetection was specific, since no staining was observed when primary antibodies were replaced by rabbit IgG (Fig 3) or when the antibodies were blocked with the respective peptides (S3 Fig).

Bottom Line: Fibroblast growth factors receptors (FGFRs) have been widely characterized in somatic cells, but there is scarce evidence of their expression and function in mammalian gametes.Incubation with FGF2 led to a significant increase in the percentage of total and progressive sperm motility, as well as in sperm kinematics.In addition to confirming the expression of FGFRs in germ cells of the human testis, our study describes for the first time the presence, localization and functionality of human sperm FGFRs, and provides evidence of the beneficial effect of FGF2 upon sperm motility.

View Article: PubMed Central - PubMed

Affiliation: Instituto de Biología y Medicina Experimental (IBYME), CONICET-FIBYME, Buenos Aires, Argentina.

ABSTRACT
Fibroblast growth factors receptors (FGFRs) have been widely characterized in somatic cells, but there is scarce evidence of their expression and function in mammalian gametes. The objective of the present study was to evaluate the expression of FGFRs in human male germ cells, to determine sperm FGFR activation by the FGF2 ligand and their participation in the regulation of sperm motility. The expression of FGFR1, 2, 3 and 4 mRNAs and proteins in human testis and localization of these receptors in germ cells of the seminiferous epithelium was demonstrated. In ejaculated sperm, FGFRs were localized to the acrosomal region and flagellum. Sperm exposure to FGF2 caused an increase in flagellar FGFR phosphorylation and activation of extracellular signal-regulated kinase (ERK) and protein kinase B (PKB or Akt) signaling pathways. Incubation with FGF2 led to a significant increase in the percentage of total and progressive sperm motility, as well as in sperm kinematics. All responses were prevented by sperm preincubation with BGJ398, a specific inhibitor of FGFR tyrosine kinase activity. In addition to confirming the expression of FGFRs in germ cells of the human testis, our study describes for the first time the presence, localization and functionality of human sperm FGFRs, and provides evidence of the beneficial effect of FGF2 upon sperm motility.

No MeSH data available.


Related in: MedlinePlus