PSX-A-24 Late-Breaking: Testosterone production by theca cells of large preovulatory follicles is affected by growth hormone depending on the hen age and the presence of granulosa cells

2021 ◽  
Vol 99 (Supplement_3) ◽  
pp. 367-368
Author(s):  
Olga V Aleynikova ◽  
Araksiya A Smekalova ◽  
Olga S Mityashova ◽  
Elena K Montvila ◽  
Irina Y Lebedeva
Keyword(s):  
Growth Hormone ◽  
Culture System ◽  
Ovarian Function ◽  
Reproductive Aging ◽  
Theca Cells ◽  
Testosterone Production ◽  
Preovulatory Follicles ◽  
Follicular Size ◽  
Spent Media

Abstract Testosterone produced by theca cells may be involved in regulating of the growth and ovulation of hen preovulatory follicles (Rangel, Gutierrez, Gen Comp Endocrinol, 203:250, 2014). In the current research, we studied effects of growth hormone (GH), a known regulator of the hen ovarian function, on in vitro testosterone production by the theca layer (TL) from the two largest yellow follicles in relation to the hen age and the presence of the granulosa layer (GL). Young hens with long clutch (YLC, 32–33 week-old, >10 eggs per clutch) and old hens with short clutch (OSC, 74–76 week-old, 3–6 eggs per clutch) were used. After isolation, TL from F1 and F2 follicles (n = 8–9) was cultured for 18 h in two systems, separately or together with the corresponding GL, in the presence or absence of chicken GH (25 ng/ml). Concentrations of testosterone in the spent media were measured by ELISA. The data were analyzed by RM-ANOVA. In the case of separate TL culture, GH did not change significantly testosterone production in both follicles of YLC hens and reduced it from 338±105 to 152±52 fmol/mg tissue (P < 0.05) in F1 follicles of OSC hens. When TL was cultured in the presence of GL, GH enhanced 1.8-2.6-fold (P < 0.05) the secretion of testosterone in the case of F1 follicles and decreased it 1.8-2.5-fold (P < 0.05) in the case of F2 follicles in both young and old hens. Regardless of the treatment, follicular size or culture system, the production of testosterone in OSC hens was 2–5 times higher than in YLC hens. The results indicate that the interaction between TL and GL changes the steroidogenic response of theca cells from preovulatory follicles to GH in young and old hens. Furthermore, testosterone production is obviously increased with reproductive aging of laying hens. The study was supported by RFBR (19-016-00216).

scholarly journals PSII-32 In vitro effect of growth hormone on progesterone production by large preovulatory follicles depends on the hen age and follicular layers interaction

2020 ◽  
Vol 98 (Supplement_4) ◽  
pp. 372-372
Author(s):  
Araksiya Smekalova ◽  
Elena K Montvila ◽  
Olga Konovalova ◽  
Olga Mityashova ◽  
Irina Lebedeva
Keyword(s):  
Growth Hormone ◽  
Repeated Measures ◽  
Ovarian Function ◽  
Culture Media ◽  
In Vitro Production ◽  
Progesterone Production ◽  
Preovulatory Follicles ◽  
Vitro Production ◽  
Vitro Effect

Abstract Growth hormone (GH) is an endocrine and paracrine/autocrine regulator of the hen ovarian function, with the GH receptor concentration in the granulosa layer (GL) being maximum in the largest preovulatory follicle (Lebedeva et al. 2004, Biol.Reprod. 71:1174–1181). In the present study, GH effects on in vitro production of progesterone by GL from the two largest yellow follicles (F1 and F2) were investigated due to the hen age and the presence of the theca layer (TL). Young hens with long clutch (YLC, 32–33 week-old, >10 eggs per clutch) and old hens with short clutch (OSC, 74–76 week-old, 3–6 eggs per clutch) were used. After isolation, GL from F1 and F2 follicles (n = 8–9) was cultured separately or jointly with the respective TL for 18 h in the presence or absence of chicken GH (25 ng/ml). Concentrations of progesterone in culture media were measured by ELISA. The data were analyzed by repeated measures ANOVA. When GL from F1 follicle cultured alone, GH did not affect progesterone production in YLC hens and decreased it from 30,5±3,4 to 20,5±2,9 ng/mg tissue (P < 0.01) in OSC hens. Conversely, when tested GL from F2 follicle, GH increased progesterone output from 15,8±2,4 to 20,4±2,5 ng/mg tissue (P < 0.05) in YLC birds and had no effect on the output in OSC birds. During co-culture of GL and TL, GH raised 1.4–1.5 times the production of progesterone in the case of F1 follicle and did not change it in the case of F2 follicle in hens of both ages. The findings indicate that the steroidogenic response of GL from the two largest preovulatory follicles to GH differs in young and old hens. However, the interaction with TL modifies the GL response and makes it similar in birds regardless the age and reproductive status. The study was supported by RFBR (19-016-00216).

Direct effects of the algal toxin, domoic acid, on ovarian function: Bovine granulosa and theca cells as an in vitro model

2015 ◽  
Vol 113 ◽  
pp. 314-320 ◽  
Author(s):  
Fabiola Pizzo ◽  
Francesca Caloni ◽  
Nicole B. Schreiber ◽  
Luis F. Schutz ◽  
Morgan L. Totty ◽  
...  
Keyword(s):  
Domoic Acid ◽  
In Vitro Model ◽  
Ovarian Function ◽  
Direct Effects ◽  
Theca Cells ◽  
Algal Toxin ◽  
Vitro Model

Tokishakuyakusan Stimulates Progesterone and Estradiol-17 β Production by Rat Granulosa Cells and Progesterone, Testosterone and Estradiol-17 β by the Residual Portion of the Follicle in Vitro

1991 ◽  
Vol 19 (02) ◽  
pp. 155-161 ◽  
Author(s):  
Satoshi Usuki
Keyword(s):  
Granulosa Cells ◽  
Cell Level ◽  
Testosterone Production ◽  
Progesterone Production ◽  
Preovulatory Follicles ◽  
Immature Rats ◽  
Serum Gonadotropin ◽  
Tissue Compartments ◽  
Stimulation Of

To examine the possible effects of Tokishakuyakusan (TS) on steroidogenesis by preovulatory follicles at the cell level, the expressed granulosa cells and remaining portion of follicles from pregnant mare's serum gonadotropin (PMS)-treated immature rats were incubated in vitro with increasing concentrations of TS for 3 h. TS significantly stimulated progesterone and estradiol-17 b production, with a predominant stimulation of progesterone, by the expressed granulosa cells, while testosterone production was not stimulated. In the remaining portion of the follicle, TS also significantly stimulated progesterone, testosterone and estradiol-17 b production. Similar to the effect produced by granulosa cells, the stimulatory effect of TS was stronger on progesterone than on testosterone and estradiol-17 b production. These results suggest that TS has a potent, direct stimulatory effect on steroidogenesis, especially progesterone production, by constituent tissue compartments of rat preovulatory follicles in vitro.

scholarly journals Spatiotemporal expression of aquaporin 9 is critical for the antral growth of mouse ovarian follicles†

2020 ◽  
Vol 103 (4) ◽  
pp. 828-839
Author(s):  
Sungeun Lee ◽  
Hee-Gyoo Kang ◽  
Chongsuk Ryou ◽  
Yong-Pil Cheon
Keyword(s):  
Chorionic Gonadotropin ◽  
Follicle Cell ◽  
Cell Types ◽  
Physiological Status ◽  
Dependent Manner ◽  
Specific Expression ◽  
Theca Cells ◽  
Spatiotemporal Expression ◽  
Preovulatory Follicles

Abstract Although a few aquaporins (AQPs) expressed in granulosa cells have been postulated to mediate fluid passage into the antrum, the specific expression of AQPs in different follicle cell types and stages and their roles have not been evaluated extensively. The spatiotemporal expression of aquaporin (Aqp) 7, 8, and 9 and the functional roles of Aqp9 in antral growth and ovulation were examined using a superovulation model and 3-dimensional follicle culture. Aqp9 was expressed at a high level in the rapid growth phase (24–48 h post equine chorionic gonadotropin (eCG) for superovulation induction) compared to Aqp7 (after human chorionic gonadotropin (hCG)) and Aqp8 (8–24 h post eCG and 24 h post hCG). A dramatic increase in the expression and localization of Aqp9 mRNA in theca cells was observed, as evaluated using quantitative reverse transcription-polymerase (RT-PCR) coupled with laser capture microdissection and immunohistochemistry. AQP9 was located primarily on the theca cells of the tertiary and preovulatory follicles but not on the ovulated follicles. In phloretin-treated mice, the diameter of the preovulatory follicles and the number of ovulated oocytes decreased. Consistent with these findings, knocking down Aqp9 expression with an Aqp9 siRNA inhibited follicle growth (0.28:1 = siRNA:control) and decreased the number of ovulated follicles (0.36:1 = siRNA:control) during in vitro growth and ovulation induction. Based on these results, the expression of AQPs is under the control of the physiological status, and AQP9 expression in theca during folliculogenesis is required for antral growth and ovulation in a tissue-specific and stage-dependent manner.

268 INFLUENCE OF DONOR AGE ON DEVELOPMENTAL COMPETENCE OF IN VITRO v. IN VIVO MATURED BOVINE OOCYTES OBTAINED BY REPEATED OVUM PICKUP FROM FOLLICLE-STIMULATING-HORMONE-STIMULATED AND NONSTIMULATED ANIMALS

2011 ◽  
Vol 23 (1) ◽  
pp. 232
Author(s):  
M. Matthiesen ◽  
H. D. Reichenbach ◽  
F. A. Habermann ◽  
M. Reichenbach ◽  
G. J. Arnold ◽  
...  
Keyword(s):  
Mixed Model ◽  
Ovarian Function ◽  
Age Groups ◽  
Developmental Competence ◽  
Reproductive Aging ◽  
Donor Age ◽  
Ovarian Aging ◽  
Bovine Oocytes

Recent findings on oogenesis, folliculogenesis, and ovarian aging in cows make the bovine system an attractive model for elucidating ovarian function and dysfunction as well as reproductive aging in women. The aim of the present study was to investigate the influence of donor age on the developmental competence of in vitro v. in vivo matured bovine cumulus–oocyte complexes (COC) obtained by ultrasound-guided repeated ovum pickup (OPU). Two groups (G1 and G2) of German Simmental heifers (14 months old at the beginning of the experiment, n = 5 and n = 7), first-lactation young cows (2–4 y old, n = 5 and n = 3), and old cows (10–15 y old, n = 5 and n = 3) were subjected to twice-weekly OPU without hormonal prestimulation 32 (G1) and 6 times (G2). Afterward, animals in G1 were punctured at 5-week intervals 9 times after FSH superstimulation to obtain in vivo matured COC at the metaphase II stage. Data were analysed using a mixed model (SAS). In the twice-weekly OPU for G1 and G2 combined, significantly (P < 0.05) more COC per animal and OPU session were obtained from the old cows (9.9 ± 1.0) compared with heifers and young cows (6.0 ± 0.8 and 7.0 ± 1.0, respectively). When G1 and G2 were regarded separately, lower numbers of COC (P < 0.01) were obtained in G1 than in G2 (2.7 ± 0.8, 4.4 ± 0.8, 7.0 ± 0.8 and 9.2 ± 1.5, 9.4 ± 2.3, 12.9 ± 2.3 for heifers, young cows, and old cows of G1 and G2, respectively). Cleavage rates (CR) on day 3 after IVF (day 0) were not affected by donor age and were not different between groups. Cultivation of COC from young cows in G1 led to higher blastocyst rates (BR) on day 7 (P < 0.05) compared with old cows and heifers. No differences in BR were observed between animals of G2. Significantly more COC (P < 0.01) were obtained in all age groups from FSH superstimulated donors (10.6 ± 0.8, 9.0 ± 0.9, and 11.7 ± 0.9 for heifers, young cows, and old cows, respectively). Cleavage rates and BR were significantly higher (P < 0.05) in all age groups after FSH superstimulation compared with those of nonstimulated donors. However, there were no differences in CR and BR between age groups (CR: 82.8 ± 7.0, 89.9 ± 7.0, 77.1 ± 6.2%; BR: 34.4 ± 7.2, 44.6 ± 7.2, 36.7 ± 7.2%). We conclude that although the numbers of COC obtained per animal and session were significantly different between G1 and G2, in vitro results were highly repeatable after OPU without hormonal prestimulation. Higher CR and BR were obtained after IVF of in vivo matured COC obtained from FSH superstimulated donors, regardless of animal age. This work was supported by the Deutsche Forschungsgemeinschaft (FOR 1041).

Growth hormone and somatostatin directly inhibit gastric ghrelin secretion. An in vitro organ culture system

2007 ◽  
Vol 30 (9) ◽  
pp. RC22-RC25 ◽  
Author(s):  
L. M. Seoane ◽  
O. Al-Massadi ◽  
F. Barreiro ◽  
C. Dieguez ◽  
F.F Casanueva
Keyword(s):  
Growth Hormone ◽  
Organ Culture ◽  
Culture System ◽  
Organ Culture System ◽  
In Vitro Organ Culture ◽  
Ghrelin Secretion

Modulating effect of growth hormone on the functional state of cultured cells from hen preovulatory follicles

2022 ◽  
pp. 108-113
Author(s):  
A. Smekalova ◽  
O. Mityashova ◽  
O. Aleinikova ◽  
E. Montvila ◽  
I. Lebedeva
Keyword(s):  
Granulosa Cells ◽  
Final Stage ◽  
Proliferative Activity ◽  
Laying Hens ◽  
Theca Cells ◽  
Apoptotic Protein ◽  
Thecal Cells ◽  
Preovulatory Follicles ◽  
Regulation Of Growth

Somatotropic hormone (STH) is an important positive modulator of ovarian function in mammals. Local production of STH and the expression of the corresponding specific receptors were also detected in hen ovarian follicles, which indicates the participation of this hormone in the endocrine/paracrine control of folliculogenesis in birds. Nevertheless, the role of STH in the regulation of growth of avian follicles at the final stage of maturation is still not clear.Objective: To study in vitro the effect of STH on the proliferative activity and apoptotic changes of granulosa and theca cells from preovulatory follicles of domestic hens.Materials and methods. Young laying hens aged 34-35 weeks with a long clutch were used in the experiments. Granulosa and theca cells were isolated from the largest yellow follicle in the hierarchy (F1). The cells were cultured in a medium containing 10% fetal bovine serum until a monolayer was formed, and then for 24 h in the medium without serum in the absence (control) or in the presence of STH at various concentrations (1-100 ng/ml). The proliferative activity and apoptotic changes in the cells were assessed by immunocytochemical assay, based on the expression level of proliferating cell nuclear antigen PCNA and pro-apoptotic protein Bax, respectively.Results. The proportion of PCNA-positive granulosa cells increased 1.3-1.8 times (P<0.01-0.05) as compared to control with increasing the content of STH in the medium to 10-100 ng/ml. Furthermore, within this concentration range, the studied hormone reduced 1.2-1.6 times (P<0.05) the relative number of granulosa cells with the positive reaction to Bax. The sensitivity of theca cells to the growth-stimulating effect of STH was lower than that of granulosa cells. Such the effect of STH led to an increase in the proportion of PCNA-positive thecal cells by 1.2-1.3 times (P<0.05) and was detected only at concentrations of 25 and 100 ng/ml. Meanwhile, STH (25-100 ng/ml) increased 1.3 times (P<0.05) the level of Bax expression in theca cells.Conclusions. The results of the present study indicate the stimulating effect of STH in vitro on the proliferative activity of granulosa and theca cells from the most mature hen preovulatory follicle. In addition, STH is able to reduce the expression of the pro-apoptotic protein Bax in granulosa cells and increase this expression in thecal cells. Thus, the data obtained indicate the possible participation of STH in the regulation of growth and development of follicles at the final stage of maturation during the period of maximum egg-laying intensity in laying hens.

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