Immunohistochemical localization of insulin-like growth factor-1 (IGF-1) in the sheep ovary and the synergistic effect of IGF-1 and FSH on follicular development in vitro and LH receptor immunostaining

2019 ◽  
Vol 129 ◽  
pp. 61-69 ◽  
Author(s):  
A.P.O. Monte ◽  
V.R.P. Barros ◽  
J.M. Santos ◽  
V.G. Menezes ◽  
A.Y.P. Cavalcante ◽  
...  
Keyword(s):  
Growth Factor ◽  
Synergistic Effect ◽  
Follicular Development ◽  
Immunohistochemical Localization ◽  
Insulin Like Growth Factor ◽  
Lh Receptor ◽  
Development In Vitro

312 EFFECTS OF TRANSFORMING GROWTH FACTOR ALPHA AND INSULIN-LIKE GROWTH FACTOR 1 SUPPLEMENTATION ON IN VITRO MATURATION OF CANINE OOCYTES

2015 ◽  
Vol 27 (1) ◽  
pp. 245
Author(s):  
A. Sato ◽  
B. Sarentonglaga ◽  
K. Ogata ◽  
M. Yamaguchi ◽  
A. Hara ◽  
...  
Keyword(s):  
Growth Factor ◽  
Synergistic Effect ◽  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
Transforming Growth Factor ◽  
Germinal Vesicle ◽  
Control Group ◽  
Insulin Like Growth Factor ◽  
Treatment Groups

Although in vitro maturation (IVM) of oocytes has been successfully established for many species, the efficiency of IVM in canine oocytes is still very low. As growth factors have been shown to promote oocyte maturation in some species, we investigated whether use of transforming growth factor α (TGF-a) and insulin-like growth factor 1 (IGF-1) might overcome the difficulties of achieving meiotic maturation in cultured canine cumulus-oocyte complexes (COC). Ovaries were obtained from bitches at 6 months to 7 years of age by ovariohysterectomy and were sliced repeatedly to release COC. In the first experiment, the COC were cultured at 38.8°C for 48 h in 5% CO2 in air in medium 199 supplemented with either TGF-a (0, 1, 10, or 100 ng mL–1) or IGF-1 (0, 0.5, 5, 10, or 50 µg mL–1). In the second experiment, the synergistic effect of TGF-a and IGF-1 was investigated by culturing COC in medium 199 supplemented with both TGF-a (0, 1, 10, or 100 ng mL–1) and IGF-1 (0, 0.5, 5, 10, or 50 µg mL–1). At the end of the culture period, the oocytes were denuded of cumulus cells by pipetting with a fine bore glass pipette; the denuded oocytes were then fixed in Carnoy's solution and stained with Hoechst 33342. The nuclear configuration and chromatin morphology of the oocytes were evaluated under confocal laser scanning microscopy. The cells were assigned to 1 of the following meiotic stages: germinal vesicle (GV), germinal vesicle breakdown (GVBD), metaphase I (MI), or metaphase II (MII). Data were analysed by ANOVA with Fisher's PLSD test. In experiment 1, no significant difference were observed in the rates of cells maturing to the MI and MII stages, but that in the 10 ng mL–1 of TGF-a group (56.3%) were larger than in the other treatment groups (38.8–51.0%). The frequencies of MII stage cells in the 5, 10, and 50 µg mL–1 of IGF-1 treatment groups (9.8, 13.3, and 12.2%, respectively) were significantly higher than in the 0.5 µg mL–1 of IGF-1 group and the control group (5.3 and 2.2%, respectively). In experiment 2, the frequency of MI and MII cells in the control, 1 ng mL–1 of TGF-a plus 0.5 µg mL–1 of IGF-1, 10 ng mL–1 of TGF-a plus 5 µg mL–1 of IGF-1, 10 ng mL–1 of TGF-a plus 10 µg mL–1 of IGF-1, and 100 ng mL–1 of TGF-a plus 50 µg mL–1 of IGF-1 group were 44.1, 36.1, 63.5, 70.8, and 50.8%, respectively. The frequency of MII cells in the control group and the same treatment groups were 2.8, 7.2, 10.4, 15.3, and 10.8%, respectively. Both frequencies in the 10 ng mL–1 of TGF-a plus 10 µg mL–1 of IGF-1 group were significantly higher than in the control group. The TGF-a may act in a paracrine fashion on the surrounding granulosa cells, and IGF-1 may play multiple roles in cellular metabolism, proliferation, growth, and differentiation in canine oocyte maturation, as has been reported for many other species. In conclusion, these results demonstrate that a synergistic effect between TGF-a and IGF-1 produces an increased rate of in vitro maturation to the MI and MII stages in canine oocytes.

129 EFFECT OF INSULIN-LIKE GROWTH FACTOR 1 AND FOLLICLE-STIMULATING HORMONE IN A DYNAMIC MEDIUM ON VIABILITY AND FOLLICULAR DEVELOPMENT OF CAPRINE EARLY PREANTRAL FOLLICLES

2012 ◽  
Vol 24 (1) ◽  
pp. 177
Author(s):  
D. Magalhães-Padilha ◽  
F. Gabriela ◽  
K. Haag ◽  
M. Gastal ◽  
K. Jones ◽  
...  
Keyword(s):  
Growth Factor ◽  
Early Stage ◽  
Follicle Stimulating Hormone ◽  
Follicular Development ◽  
Blue Dye ◽  
Insulin Like Growth Factor ◽  
Preantral Follicles ◽  
Secondary Follicle

The goal of this study was to investigate the effect of a dynamic medium supplemented with insulin-like growth factor 1 (IGF-1), FSH, or both on the viability, activation and secondary follicle rates and on the follicle and oocyte diameters of caprine preantral follicles submitted to a long-term (16 days of culture) in vitro culture system. Fragments from goat ovaries (n = 16) obtained from slaughterhouse were cultured in α-MEM containing or not containing IGF-1 (50 ng mL–1), FSH (50 ng mL–1), or both as a dynamic medium that was added in the first (Day 0 to 8) and second (Day 8 to 16) halves of culture, resulting in 6 treatments: α-MEM alone, IGF-1/IGF-1, FSH/FSH, IGF-1/FSH, FSH/IGF-1 and IGF-1 + FSH/IGF-1 + FSH. Noncultured (fresh control) and cultured fragments were processed for morphological and viability analyses. Follicles within ovarian fragments were mechanically isolated and early-stage follicles were classified as normal or abnormal and primordial, primary, or secondary. The viability of isolated follicles was determined by trypan blue dye and follicles were classified as live or dead. For this experiment, 6240 preantral follicles were analysed. Data for statistical analyses were transformed and submitted to ANOVA using the GLM procedure of SAS, followed by the Duncan test for comparison of means. At Day 8 of culture, more (P < 0.05) follicles in the treatments containing IGF-1 alone or associated with FSH were normal and viable than in the treatments cultured with FSH or α-MEM alone. At Day 16 of culture, the highest (P < 0.05) percentage of viability was observed in the IGF-1/IGF-1 (68%), IGF-1/FSH (68%) and IGF-1 + FSH/IGF-1 + FSH (72%) treatments. However, more (P < 0.05) normal follicles were observed on Day 16 in the IGF-1 + FSH/IGF-1 + FSH treatment (76%) than in all other treatments, except for the IGF-1/FSH treatment (72%). The percentage of follicular activation (primordial to primary) increased (P < 0.05) in all treatments from Day 0 to 8 (mean, 5 to 49%, respectively). The rate of follicular activation increased (P < 0.05) from Day 8 to 16 in all groups, except for the FSH/FSH and IGF-1 + FSH/IGF-1 + FSH treatments. Nevertheless, at Day 16, the IGF-1 + FSH/IGF-1 + FSH treatment had the highest (P < 0.05) percentage of secondary follicles (28%) when compared with the other groups (range, 6 to 17%). Furthermore, the IGF-1 + FSH/IGF-1 + FSH treatment had the largest (P < 0.05) mean follicular and oocyte diameters after 16 days of culture. In summary, follicular morphology and viability were maintained, follicle activation was promoted and secondary follicle formation was stimulated with the association of IGF-1 and FSH. Therefore, our results demonstrate the importance of IGF-1 associated with FSH during the entire long-term culture period on early folliculogenesis in goats.

scholarly journals Changes in insulin-like growth factor binding protein (IGFBP) isoforms during bovine follicular development

Reproduction ◽  
2002 ◽  
pp. 439-446 ◽  
Author(s):  
B Nicholas ◽  
RK Scougall ◽  
DG Armstrong ◽  
R Webb
Keyword(s):  
Growth Factor ◽  
Follicular Fluid ◽  
Follicular Development ◽  
Immunoblot Analysis ◽  
Proteolytic Cleavage ◽  
Insulin Like Growth Factor ◽  
Principal Mechanism ◽  
Post Translational Modifications ◽  
Factor Binding

UThe insulin-like growth factor binding proteins (IGFBPs) bind IGFs with high affinity and so regulate their access to the type 1 and 2 IGF receptors. This is the principal mechanism involved in regulating IGF bioavailability during folliculogenesis. IGFBPs undergo a number of post-translational modifications, including proteolytic cleavage, phosphorylation and glycosylation, which can regulate the affinity of IGFBPs for IGFs. However, the post-translational changes to IGFBPs that occur during folliculogenesis have not been fully characterized. The charge and size variants of the IGFBPs in bovine follicular fluid were examined by two-dimensional non-reducing SDS-PAGE followed by non-isotopic western ligand blot analysis, and immunoblot analysis during follicular development. The results demonstrate the presence of at least 51 IGFBP isoforms corresponding to IGFBP-1 to -6 in bovine follicular fluid from subordinate follicles, many of which were phosphorylated. The total number of IGFBPs was reduced in dominant follicles, whereas no gross changes in isoforms were observed during follicular development. These results demonstrate the high degree of conservation of IGFBP post-translational modifications between species, and from the in vitro dephosphorylation of these proteins it is hypothesized that these modifications may result in changes to IGF binding or susceptibility to proteolytic cleavage.

scholarly journals Identification of the types of insulin-like growth factor-binding proteins that are secreted by muscle cells in vitro

1989 ◽  
Vol 264 (14) ◽  
pp. 7795-7800
Author(s):  
R H McCusker ◽  
C Camacho-Hübner ◽  
D R Clemmons
Keyword(s):  
Growth Factor ◽  
Binding Proteins ◽  
Muscle Cells ◽  
Insulin Like Growth Factor ◽  
Factor Binding

Blended Effects of Herbal Components of Tokishakuyakusan on Somatomedin C/Insulin-like Growth Factor 1 Level in Rat Corpus Luteum

1991 ◽  
Vol 19 (01) ◽  
pp. 61-64 ◽  
Author(s):  
Satoshi Usuki
Keyword(s):  
Growth Factor ◽  
Corpus Luteum ◽  
Corpora Lutea ◽  
Insulin Like Growth Factor ◽  
Somatomedin C ◽  
Factor I ◽  
Peony Root ◽  
Growth Factor I ◽  
Herbal Components

The effect of herbal components of Tokishakuyakusan on somatomedin C/insulin-like growth factor I (IGF-1) level in medium from rat corpora lutea incubated in vitro was examined. Hoelen + peony root + Japanese angelica root, hoelen + peony root, hoelen + Japanese angelica root or peony root + Japanese angelica root decreased the IGF-1 level. The data suggest that constituent herbal components of Tokishakuyakusan regulate the IGF-1 level by rat corpora lutea.

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