Effect of Exogenous Melatonin on the Development of Mice Ovarian Follicles and Follicular Angiogenesis

In mammalian, the periodic growth and development of ovarian follicles constitutes the physiological basis of female estrus and ovulation. Concomitantly, follicular angiogenesis exerts a pivotal role in the growth of ovarian follicles. Melatonin (N-acetyl-5-methoxytryptamine, Mel), exists in follicle fluid, was suggested to affect the development of follicles and angiogenesis. This research was conducted to investigate the effects and mechanisms of Mel on the development of ovarian follicles and its angiogenesis. In total, 40 ICR mice at age of 3 weeks were allocated into four groups at liberty: control, Mel, FSH and FSH + Mel for a 12-day trial. Ovaries were collected at 8:00 a.m. on Day 13 for detecting the development of ovarian follicles and angiogenesis. Results indicated that Mel promoted the development of ovarian follicles of 50–250 μm (secondary follicles) and periphery angiogenesis, while FSH remarkably increased the number of antral follicles and periphery angiogenesis. Mechanically, Mel and FSH may regulate the expression of VEGF and antioxidant enzymes in different follicular stages. In conclusion, Mel primarily acted on the secondary follicles, while FSH mainly promoted the development of antral follicles. They both conduced to related periphery angiogenesis by increasing the expression of VEGF. These findings may provide new targets for the regulating of follicular development.

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Ontogeny of the long form of leptin receptor gene expression in the porcine ovarian follicles

Polish Journal of Veterinary Sciences ◽

10.2478/pjvs-2013-0014 ◽

2013 ◽

Vol 16 (1) ◽

pp. 101-105

Author(s):

N. Smolinska ◽

T. Kaminski ◽

G. Siawrys ◽

J. Przala

Keyword(s):

Energy Homeostasis ◽

Leptin Receptor ◽

Receptor Gene ◽

Follicular Development ◽

Ovarian Follicles ◽

Antral Follicles ◽

Long Form ◽

Rb Gene ◽

Receptor Gene Expression

Abstract Leptin is a polypeptide hormone produced predominantly in adipocytes. It has been found to be implicated in the regulation of satiety and energy homeostasis. A role for leptin in reproduction was later suggested by findings that this hormone may be involved in the regulation of the hypothalamic- pituitary-gonadal axis via endocrine, paracrine and/or autocrine pathways. The objective of the study was to investigate the ontogeny of the long isoform of leptin receptor (OB-Rb) gene in porcine ovarian follicles. The expression of OB-Rb gene was detected in porcine primordial, primary, secondary and antral follicles by in situ hybridization. In summary, our data suggest that leptin might have a direct effect on porcine follicles and plays an important role in the follicular development.

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Development and survivability of ovarian follicles of goat in different feeding systems

Bangladesh Journal of Animal Science ◽

10.3329/bjas.v43i3.21644 ◽

2014 ◽

Vol 43 (3) ◽

pp. 175-179

Author(s):

MH Alam ◽

ME Kabir ◽

MB Sarker ◽

BK Saha ◽

RI Khan ◽

...

Keyword(s):

Histological Examination ◽

Morphometric Analysis ◽

Follicular Development ◽

Ovarian Follicles ◽

Primordial Follicles ◽

Antral Follicles ◽

Feeding Systems

The present study was aimed to know the effects of feeding systems on follicular development in Black Bengal goats. Nine female post-weaned Black Bengal goats were randomly assigned to 3 feeding systems namely stall feeding, tethering and grazing. After 240 days of rearing, goats were slaughtered and ovaries were collected for morphometric analysis. Histological examination revealed that the percentages of secondary (p<0.01) and antral follicles (p<0.05) were higher in stall-fed than tethering and grazing goats. Percentage of primordial follicles (61±2%) was lower in stall-fed goats and higher in tethering (71±3%) goats. Number of degenerated follicles was higher in tethering goats and lower in stall-fed groups. These results reveal that development and degeneration of ovarian follicles are influenced by feeding systems of goats.DOI: http://dx.doi.org/10.3329/bjas.v43i3.21644 Bang. J. Anim. Sci. 2014. 43 (3): 175-179

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Leptin Supplementation Stimulates Synergism with Growth Factors and Hormones to Express Its Receptor in Cultured Preantral Follicles of Sheep

Indian Journal of Animal Research ◽

10.18805/ijar.b-4218 ◽

2020 ◽

Author(s):

K. Sravani Pragna ◽

V. Praveen Chakravarthi ◽

Deepa Pathipati ◽

B. Rambabu Naik ◽

L.S.S. Varaprasad Reddy ◽

...

Keyword(s):

Growth Factors ◽

Leptin Receptor ◽

Culture Media ◽

Follicular Development ◽

Cumulus Cells ◽

Ovarian Follicles ◽

Antral Follicles ◽

Receptor Mrna

Background: Leptin receptor is a transmembrane receptor that regulates reproduction at molecular level.Since for action of any hormone on target cell and to have local action on any tissue, expression of its own receptor is necessary and also it is not known whether such improvement in ovarian follicular development by Leptin is mediated through presence of its homologous receptors in the sheep ovaries. Therefore this study aimed on expression of Leptin receptor mRNA in cultured ovarian follicles of sheep by RT PCR.Methods: Leptin receptor mRNA expression in sheep was studied using qRT-PCR from: (i) In vivo grown preantral, early antral, antral, large antral follicles and cumulus oocyte complexes obtained from large antral follicles subjected to 24h of in vitro maturation and (ii) PFs’ exposed to three different culture media for 3min, two, four or six days and subsequently matured in vitro for 24h. Result: Leptin receptor was observed at all stages ovarian follicles in both cumulus cells and oocytes. Leptin supplementation along with other growth factors and hormones stimulated the expression of its receptor mRNA which is parallel to in vivo stages which could suggest synergistic action of growth factors and hormones with Leptin.

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The effect of acute immuno-neutralisation of inhibin in ewes during the early luteal phase of the oestrous cycle on ovarian hormone secretion and follicular development

Journal of Endocrinology ◽

10.1677/joe.0.1450479 ◽

1995 ◽

Vol 145 (3) ◽

pp. 479-490 ◽

Cited By ~ 13

Author(s):

B K Campbell ◽

B M Gordon ◽

C G Tsonis ◽

R J Scaramuzzi

Keyword(s):

Luteal Phase ◽

Follicular Development ◽

Experimental Period ◽

Ovarian Follicles ◽

Passive Immunisation ◽

Follicle Development ◽

Ovarian Steroid ◽

Blood Samples ◽

Steroid Secretion ◽

Antibody Titres

Abstract Ewes with ovarian autotransplants received either inhibin antiserum (10 ml i.v. raised in sheep against recombinant 32 kDa human inhibin; n=6) or sheep serum (10 ml i.v.; n=5) on day 3 of the luteal phase with additional daily injections (1 ml i.v.) from 48 h after the initial bolus until day 13. Jugular and ovarian venous blood samples were taken 4-hourly over days 2–13 of the luteal phase. Blood samples were also taken at more frequent intervals (every 10–15 min for 2–3 h) to examine pulsatile secretory responses from the ovary to endogenous and gonadotrophin-releasing hormone-induced (150 ng i.m.) LH pulses on days 4, 6, 8, 10 and 12 of the luteal phase. Plasma FSH levels, ovarian steroid secretion and ovarian follicular development were measured. The ovarian follicle population was estimated daily by real time ultrasound scanning. Immunisation against inhibin resulted in a 3- to 4-fold increase (P<0·001) in plasma FSH levels within 8 h with levels remaining elevated over controls for 6–7 days. Within 24 h of immunisation there was an increase in the number of small ovarian follicles (P<0·05) and by 3 days after treatment immunised ewes had 4–6 large ovarian follicles/ewe with this increase in the total number of large follicles being maintained for the rest of the experimental period (P<0·05). Mean ovarian oestradiol secretion during intensive bleeds was not different from controls 24 h after immunisation, but by 3 days after immunisation it was elevated 4- to 5-fold (P<0·001) over controls with this increase being maintained throughout the experiment. Similar responses to immunisation against inhibin in androstenedione secretion were observed although mean androstenedione secretion was not elevated until 7 days after treatment. In vitro antibody titres in immunised ewes remained elevated but declined steadily (P<0·001) over the experimental period. We conclude that the initial stimulation of follicle development and ovarian steroid secretion following passive immunisation against inhibin can be attributed to increased blood FSH. However, the fact that with time FSH declined but increased follicle development was sustained, despite maintenance of high circulating antibody titres, suggests that on a longer term basis inhibin immunisation may stimulate ovarian function by interfering with the modulation of follicle development by inhibin at an ovarian level. Journal of Endocrinology (1995) 145, 479–490

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The effects of IGF-I on bovine follicle development and IGFBP-2 expression are dose and stage dependent

Reproduction ◽

10.1530/rep.1.00682 ◽

2006 ◽

Vol 131 (3) ◽

pp. 515-523 ◽

Cited By ~ 30

Author(s):

Kirsty A Walters ◽

John P Binnie ◽

Bruce K Campbell ◽

David G Armstrong ◽

Evelyn E Telfer

Keyword(s):

Growth Factor ◽

Granulosa Cells ◽

Size Range ◽

Regulatory System ◽

Follicular Development ◽

High Dose ◽

Insulin Like Growth Factor ◽

High Concentration ◽

Antral Follicles ◽

Igf I

This study aimed to determine the effect of insulin-like growth factor-I (IGF-I) on early antral bovine follicular development, and the expression of insulin-like growth factor-binding protein-2 (IGFBP-2). Antral follicles separated into three different size groups were cultured for 6 days in medium supplemented with either a low (10 ng/ml) or high (1 μg/ml) dose of human recombinant IGF-I. Oestradiol production by follicles in all size ranges, cultured in the presence of the high concentration of IGF-I, significantly increased by day 6 (P < 0.05). Follicles in the smallest size range, 165–215 μm, cultured in a high dose of IGF-I, were found to be significantly increased in size (P < 0.01). Oocyte health of the largest follicles (281–380 μm) was significantly improved by the addition of IGF-I to the culture medium. mRNA expression of IGFBP-2 was decreased in the granulosa cells of follicles, size range 216–280 μm, cultured with a high dose of IGF-I (P < 0.05). Granulosa cells (P < 0.05) and oocytes (P < 0.01) of the largest follicles (281–380 μm) showed a decrease in IGFBP-2 expression (protein) when cultured in the control and low-IGF-I treatment groups. Therefore, the response of a bovine follicle to IGF-I is both dose and stage dependent. This work supports a role for IGF-I in modulating somatic and germ-cell maturation and development in early antral follicles. Furthermore, the inverse relationship between the level of IGF-I stimulation and IGFBP-2 expression suggests a local regulatory system modulating IGF-I availability.

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Molecular cloning, expression analysis and developmental changes in ovarian follicles of goose 3β-hydroxysteroid dehydrogenase 1

Animal Production Science ◽

10.1071/an13315 ◽

2014 ◽

Vol 54 (8) ◽

pp. 992 ◽

Cited By ~ 3

Author(s):

Yingying Zhang ◽

Hehe Liu ◽

Mingjun Yang ◽

Shengqiang Hu ◽

Liang Li ◽

...

Keyword(s):

Adrenal Gland ◽

Follicular Development ◽

Hydroxysteroid Dehydrogenase ◽

Ovarian Follicles ◽

Mrna Levels ◽

Rt Pcr ◽

Chain Reaction ◽

Polymerase Chain ◽

Human And Mouse ◽

Main Factor

The enzyme 3β-hydroxysteroid dehydrogenase/isomerase1 (3βHSD1) can catalyse the conversion of pregnenolone to progesterone in the △4-3-ketosteroid metabolic pathway. The aim of the present study was to clone 3βHSD1 and to determine whether this enzyme in the follicular wall has an effect on yolk progesterone in geese (Anser cygnoides). A putative coding sequence of 3βHSD1, which was 1134 nucleotides in length, was successfully obtained by using reverse transcription polymerase chain reaction (RT–PCR). A comparison of the deduced amino acid sequence with chicken, quail, zebra finch, cattle, horse, pig, human and mouse 3βHSD1 showed 89.7%, 88.4%, 87.3%, 55.6%, 54.0%, 53.5%, 55.3% and 52.9% similarity, respectively. The detection of 3βHSD1 mRNA levels in several tissues by quantitative real-time PCR showed that the highest level of 3βHSD1 was in the adrenal gland, followed by the ovary, which indicated that the gene we obtained was the adrenal gland/gonad-specific one. We measured the level of 3βHSD1 mRNA in the follicular wall and determined the concentration of progesterone in the yolk of these ovarian follicles; the concentration of progesterone in the yolk had a pattern of expression similar to that of 3βHSD1 in the follicular wall during follicular development. This result suggests that the expression of 3βHSD1 in the follicular wall may be a main factor that contributes to the accumulation of yolk progesterone.

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Effects of species differences on oocyte regulation of granulosa cell function

Reproduction ◽

10.1530/rep-12-0267 ◽

2012 ◽

Vol 144 (5) ◽

pp. 557-567 ◽

Cited By ~ 20

Author(s):

Jia Yi Lin ◽

Janet L Pitman-Crawford ◽

Adrian H Bibby ◽

Norma L Hudson ◽

C Joy McIntosh ◽

...

Keyword(s):

Growth Factors ◽

Species Differences ◽

Cell Function ◽

Thymidine Incorporation ◽

Follicular Development ◽

Specific Antibodies ◽

Antral Follicles ◽

Morphogenetic Protein ◽

Bone Morphogenetic Protein 15 ◽

Highly Correlated

The aims were to investigate whether oocyte-secreted growth factors from a high (i.e. rat) and low (i.e. sheep) ovulation rate species could stimulate 3H-thymidine incorporation in granulosa cells (GC) from antral follicles from the same or across species. Denuded oocytes (DO) were co-incubated with GC with or without specific antibodies to growth differentiating factor 9 (GDF9) or bone morphogenetic protein 15 (BMP15). Co-incubations of DO-GC from the same or across species significantly increased thymidine incorporation in GC with increasing numbers of DO. GDF9 immuno-neutralisation reduced thymidine incorporation in rat GC co-incubated with either rat or ovine DO and in ovine GC co-incubated with ovine or rat DO. BMP15 immuno-neutralisation only reduced thymidine incorporation when ovine DO were co-incubated with either ovine or rat GC. Western blotting of oocytes co-incubated with GC identified GDF9 and BMP15 proteins for sheep and GDF9 protein for rats in oocyte lysates and incubation media. With respect to rat BMP15, a promature protein was identified in the oocyte lysate but not in media. Expression levels of GDF9 relative to BMP15 mRNA in DO co-incubated with GC were highly correlated (R 2=0.99) within both species. However, the expression ratios were markedly different for the rat and sheep (4.3 vs 1.0 respectively). We conclude that during follicular development, rat oocytes secrete little, if any, BMP15 and that GDF9 without BMP15 can stimulate proliferation of rat and ovine GC. In contrast, ovine oocytes secrete both BMP15 and GDF9, and both were found to stimulate proliferation in ovine and rat GC.

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Follicular development, morphological integrity, and oxidative stress in bovine preantral follicles cultured in vitro with ascorbic acid

Zygote ◽

10.1017/s0967199421000903 ◽

2021 ◽

pp. 1-7

Author(s):

Larissa Zamparone Bergamo ◽

Denis Vinicius Bonato ◽

Camila Bizarro-Silva ◽

Francieli Gesleine Capote Bonato ◽

Tamires Korchovei Sanches ◽

...

Keyword(s):

Oxidative Stress ◽

Ascorbic Acid ◽

Culture Medium ◽

Bos Taurus ◽

Follicular Development ◽

Ovarian Follicles ◽

Bos Taurus Indicus ◽

Bonferroni Test ◽

And Oxidative Stress

Summary The aim of this study was to evaluate the follicular development, morphological integrity, and oxidative stress of preantral ovarian follicles from Bos taurus indicus females grown in vitro with ascorbic acid. Ovaries (n = 20) from Bos taurus indicus females were collected, fragmented, and were cultured in vitro for 6 or 12 days in minimum essential medium (MEM), or MEM supplemented with 50 or 100 ng/ml ascorbic acid, with an extracellular matrix of agarose gel, in an incubator at 38.5°C; every 2 days, 100% of the culture medium was replaced. The data were analyzed using the chi-squared test and/or Fisher’s exact test. In the event of a significant effect, the proportions were compared using a 2 × 2 proportion test. The oxidative stress analysis data were submitted to analysis of variance followed by the Bonferroni test. Values were considered significant when P ≤ 0.05. The addition of 100 ng/ml of ascorbic acid to the in vitro culture medium of preantral ovarian follicles from bovine females promoted follicular development, was efficient in maintaining morphological integrity, as well as the stability of reactive oxygen species, after 6 days of in vitro culture.

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Establishment and Functional Characterization of Immortalized Rabbit Dermal Papilla Cell Lines

10.21203/rs.3.rs-1105281/v1 ◽

2021 ◽

Author(s):

Jiali Li ◽

Bohao Zhao ◽

Chen Zhang ◽

Xiyu Zhang ◽

Yingying Dai ◽

...

Keyword(s):

Cell Lines ◽

Growth And Development ◽

Functional Characterization ◽

Dermal Papilla ◽

Early Passage ◽

Dermal Papilla Cells ◽

Viability Analysis ◽

Periodic Growth ◽

Basic Characteristics ◽

Immortalized Cell

Abstract Background Hair follicle (HF) undergo periodic growth and development in mammals, which regulated by dermal papilla cells (DPCs) are reported to play an important role in the HF morphogenesis and development. However, primary DPCs have low proliferative activity, age quickly, and fresh cell isolation is both time-consuming and laborious. Method In this study, we introduced SV40LT into dissociated early passage rabbit vibrissae DPCs with lentiviral vectors and established seven immortalized DP cell lines (R-1, R-2, R-3, R-4, R-5, R-6 and R-7). Result These cell lines displayed early passage morphology and displayed high alkaline phosphatase activity. RT-PCR and immunofluorescence staining showed that all the immortalized cell lines expressed the DPC markers (α-SMA ,IGF1, ALPL, FGF2, BMP2 and TGFβ2; α-SMA and VIM protein), but α-SMA was only expressed well in R-3, R-4, and R-7. Furthermore, it was found that R-7 was the only line to survive beyond 50 passages. Compared to melanoma cells, R-7 did not undergo malignant transformation. Karyotyping and cell growth viability analysis illustrated that the R-7 cell line preserved the basic characteristics of primary DPCs. Conclusion The R-7 DPCs established have potential application for future hair research. The study provides the theoretical basis in the cell research of HF growth and development.

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Altrenogest affects the development and endocrine milieu of ovarian follicles in prepubertal and mature gilts†

Biology of Reproduction ◽

10.1093/biolre/ioaa136 ◽

2020 ◽

Vol 103 (5) ◽

pp. 1069-1084

Author(s):

Adam J Ziecik ◽

Klaudia Drzewiecka ◽

Katarzyna Gromadzka-Hliwa ◽

Jan Klos ◽

Patrycja Witek ◽

...

Keyword(s):

Follicular Fluid ◽

Messenger Rna ◽

Ovarian Follicle ◽

Adenosine Monophosphate ◽

Cyst Formation ◽

Ovarian Follicles ◽

Mrna Levels ◽

Molecular Control ◽

Antral Follicles ◽

Follicular Cyst

Abstract Altrenogest with gonadotropins is commonly used to synchronize the estrous cycle, but it can also lead to follicular cyst formation, especially in prepubertal gilts. Here, we aimed to investigate how maturity and altrenogest treatment affect the development, endocrine milieu, and molecular control of ovarian follicles. Crossbred prepubertal and mature gilts were challenged or not (control) with altrenogest, and ovaries were collected in the morning on the first day of behavioral estrus. In prepubertal gilts, altrenogest decreased the percentage of primordial and atretic small follicles, but increased large antral follicles when compared with controls. In mature gilts, altrenogest reduced the percentage of primary follicles and elevated the total number of antral follicles. Maturity affected the estradiol level in the follicular fluid of preovulatory follicles, luteinizing hormone (LH)-stimulated cyclic adenosine monophosphate (cAMP) generation, and LH receptor messenger RNA (mRNA) expression in granulosa. Moreover, cytochrome P45017A1 (CYP17A1) mRNA levels in the theca layer were affected and correlated with follicular androstendione and estradiol concentration. Altrenogest negatively affected follicular fluid progesterone concentration and decreased levels of prostaglandin (PG) E2 in prepubertal gilts and PGF2alpha metabolite in mature gilts. LH-stimulated cAMP release in granulosa cells of mature gilts as well as human chorionic gonadotropin- and forskolin-induced cAMP were also affected. In addition, altrenogest downregulated CYP17A1 mRNA in the prepubertal theca layer and PGF2alpha synthase expression in the granulosa and theca layer of mature gilts. To the best of our knowledge, this is the first study to report multiple effects of maturity and altrenogest on the endocrine milieu and molecular regulations governing ovarian follicle development in gilts.

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