Sequential effects of FSH on the first stages of ovarian follicular development in normal and dwarf Snell mice

1988 ◽  
Vol 117 (1) ◽  
pp. 26-32 ◽  
Author(s):  
M. M. de Reviers
Keyword(s):  
Cross Section ◽  
Granulosa Cells ◽  
Thymidine Incorporation ◽  
Tritiated Thymidine ◽  
Follicular Development ◽  
Sequential Effects ◽  
Mouse Ovary ◽  
Major Increase ◽  
Dwarf Mice ◽  
Stimulation Of

Abstract. The population of small growing ovarian follicles was divided into 4 classes according to the number of granulosa cells (from 15 to 95) surrounding the oocyte, and a comparison was made of normal and dwarf mice. Follicular cell proliferation was estimated by tritiated thymidine incorporation. In normal mice, most follicles in classes 1 (15 to 35 granulosa cells in their largest cross-section) and 2 (36 to 55 cells) were labelled (86 and 95%, respectively); FSH treatment increased the labelling index (L.I.) in all follicle classes. In dwarf mice, only 38 and 76% of follicles in classes 1 and 2, respectively, were labelled. However, FSH treatment increased the percentage of labelled follicles and the L.I. to levels which were similar to those in the ovaries of untreated, normal animals. FSH stimulation of the percentage of labelled follicles and L.I. was obvious as early as 3 h after injection. There was a major increase of the L.I. 24 h after FSH stimulation, specially in dwarf mice; several hypotheses are proposed to explain this finding. We conclude that FSH is necessary for the development of the population of small growing follicles in the mouse ovary.

Ultrastructural localisation of calcium deposits in the mouse ovary

2003 ◽  
Vol 15 (8) ◽  
pp. 415 ◽  
Author(s):  
M. Sedmíková ◽  
R. Rajmon ◽  
J. Petr ◽  
M. Vaňková ◽  
J. Rozinek ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Follicular Development ◽  
Mouse Ovary ◽  
Antral Follicles ◽  
Mouse Oocytes ◽  
Adult Mice ◽  
Calcium Deposits ◽  
Cellular Compartments ◽  
Meiotic Competence

Follicle-enclosed mouse oocytes contain numerous calcium deposits. The ultrastructural distribution of calcium deposits in the nuclei, mitochondria and cytoplasm of mouse oocytes and granulosa cells of primary, secondary and antral follicles was examined using the combined oxalate–pyroantimonate method. The mitochondria of oocytes from all types of follicles had the highest levels of calcium deposits of all oocyte compartments, with the exception of primary follicles, in which oocyte nuclei contained the same level of calcium deposits as the mitochondria. Calcium deposits in the cytoplasm of oocytes from primary follicles were significantly lower than those in the cytoplasm of oocytes from secondary and antral follicles. Calcium deposits in the cytoplasm of granulosa cells were significantly lower than calcium deposits in the mitochondria of granulosa cells and this difference persisted throughout all categories of follicles. Calcium deposits in the nuclei of granulosa cells did not differ from levels in the mitochondria in primary and secondary follicles. In contrast, the nuclei of granulosa cells from antral follicles had lower levels of calcium deposits than the mitochondria. The differences observed in calcium deposits in various cellular compartments in oocytes and granulosa cells in the follicles of ovaries of adult mice can be attributed to their acquisition of meiotic competence and follicular development.

187 EXPRESSION AND REGULATION OF THE FIBROBLAST GROWTH FACTOR GENE FAMILY DURING MOUSE FOLLICULAR DEVELOPMENT

2013 ◽  
Vol 25 (1) ◽  
pp. 243
Author(s):  
S. Furukawa ◽  
K. Naito ◽  
K. Sugiura
Keyword(s):  
Gene Family ◽  
Granulosa Cells ◽  
Follicular Development ◽  
Cumulus Cells ◽  
Mrna Levels ◽  
Fibroblast Growth ◽  
Mouse Ovary ◽  
Expression Levels ◽  
Before And After ◽  
Equine Chorionic Gonadotropin

Recent studies have shown the critical roles of fibroblast growth factors (FGFs), including FGF8 produced by oocytes, in regulating follicular development. However, the expression and regulation of the FGF gene family, which consists of 22 ligands and 4 receptors, in the mouse ovary have not been well understood. The aim of the present study was to assess the expression and regulation of FGF ligands and receptors in the mouse ovary. Transcript levels of FGF ligands and receptors in immature (3-week-old) and adult (7- to 8-week-old) ovaries as well as other tissues of B6/DBA2F1 mice were analysed with RT-PCR. Furthermore, expression levels of FGF receptors in cumulus cells (CC) and mural granulosa cells (MG) before and after equine chorionic gonadotropin (eCG) treatment were determined with RT-quantitative PCR. Among 21 FGF ligands examined, 12 and 9 transcripts were detectable in immature and adult ovaries, respectively. More FGF ligands were detected in ovary, testis, heart, and brain compared to other tissues, including liver and spleen. Transcripts of all 4 FGF receptors (Fgfr1–4) were detectable in both immature and adult ovaries. Expression levels of Fgfr1 and Fgfr2 were significantly higher in MG compared with CC before and after the eCG treatment. Levels of Fgfr4 were comparable between MG and CC before the eCG treatment, but became significantly different with higher expression levels in MG after the eCG treatment. Fgfr3 transcripts were barely detectable in CC and MG. Overall levels of Fgfr1 in granulosa cells (CC and MG) were downregulated by eCG treatment, whereas those of Fgfr2 and Fgfr4 were upregulated. In summary, many FGF ligands are expressed, at least in mRNA levels, in mouse ovaries. Moreover, the expression levels of Fgfr transcripts in granulosa cells are dynamically regulated during follicular development.

scholarly journals Gonadotropin Stimulation of Ovarian Fractalkine Expression and Fractalkine Augmentation of Progesterone Biosynthesis by Luteinizing Granulosa Cells

Endocrinology ◽  
2008 ◽  
Vol 149 (6) ◽  
pp. 2782-2789 ◽  
Author(s):  
Ping Zhao ◽  
Ananya De ◽  
Zeng Hu ◽  
Jing Li ◽  
Sabine M. Mulders ◽  
...  
Keyword(s):  
Real Time ◽  
Granulosa Cells ◽  
Regulatory Protein ◽  
Rt Pcr ◽  
Paracrine Factors ◽  
Transcript Levels ◽  
Preovulatory Follicles ◽  
Major Increase ◽  
Steroidogenic Acute Regulatory ◽  
Stimulation Of

Recent studies indicated that ovarian functions are regulated by diverse paracrine factors induced by the preovulatory increases in circulating LH. Based on DNA microarray analyses and real-time RT-PCR, we found a major increase in the transcript levels of a chemokine fractalkine after human chorionic gonadotropin (hCG) treatment during the preovulatory period in gonadotropin-primed immature mice and rats. Although CX3CR1, the seven-transmembrane receptor for fractalkine, was also found in murine ovaries, its transcripts displayed minimal changes. Using tandem RT-PCR and immunohistochemistry, fractalkine transcripts and proteins were localized in cumulus, mural granulosa, and theca cells as well as the oocytes, whereas CX3CR1 was found in the same cells except the oocyte. Real-time RT-PCR further indicated the hCG induction of fractalkine transcripts in different ovarian compartments, with the highest increases found in granulosa cells. In cultured granulosa cells, treatment with fractalkine augmented hCG stimulation of progesterone but not estradiol and cAMP biosynthesis with concomitant increases in transcript levels for key steroidogenic enzymes (steroidogenic acute regulatory protein, CYP11A, and 3β-hydroxysteroid dehydrogenase). In cultured preovulatory follicles, treatment with fractalkine also augmented progesterone production stimulated by hCG. Furthermore, treatment with fractalkine augmented the phosphorylation of P38 MAPK in cultured granulosa cells. The present data demonstrated that increases in preovulatory LH/hCG induce the expression of fractalkine to augment the luteinization of preovulatory granulosa cells and suggest the fractalkine/CX3CR1 signaling system plays a potential paracrine/autocrine role in preovulatory follicles.

FOLLICLE GROWTH IN THE IMMATURE MOUSE OVARY

1969 ◽  
Vol 62 (1) ◽  
pp. 117-132 ◽  
Author(s):  
Torben Pedersen
Keyword(s):  
Granulosa Cells ◽  
Tritiated Thymidine ◽  
List Type ◽  
Pulse Labelling ◽  
Follicle Growth ◽  
Mouse Ovary ◽  
Stage Of Development ◽  
Different Ages ◽  
Immature Mouse ◽  
Doubling Times

ABSTRACT The growth of follicles in the immature mouse ovary was investigated in autoradiographs prepared after pulse-labelling with tritiated thymidine. Three parameters, which determine follicle growth were estimated: The number of follicles present in the ovary at different ages. The time it takes follicles to grow from one stage of development to another. This was calculated from the total number of granulosa cells in these stages and from their doubling times. The doubling time of granulosa cells was determined from their labelling index and the duration of their DNA-synthesis phase. The number of follicles, which start on their development at different ages. It was found, that the follicle development is not constant in the period from birth to maturity, but varies considerably. More follicles start to grow in the young than in the older immature mouse. Moreover the follicles grow faster early in life than later. The development from a follicle with one layer of granulosa cells to one with several layers and antrum formation takes about 10 days in the first half of the immature period, while it takes about 16 days as the animal approaches maturity. It was furthermore shown, that about 850 follicles start to grow in the immature period.

Influence of growth factors on the plasminogen activator activity of avian granulosa cells from follicles at different maturational stages of preovulatory development

1993 ◽  
Vol 11 (3) ◽  
pp. 291-304 ◽  
Author(s):  
M Lafrance ◽  
F Croze ◽  
B K Tsang
Keyword(s):  
Growth Factors ◽  
Growth Factor ◽  
Plasminogen Activator ◽  
Granulosa Cells ◽  
Tritiated Thymidine ◽  
Follicular Development ◽  
Mitogenic Action ◽  
Preovulatory Follicles ◽  
Epidermal Growth ◽  
Assay Methods

ABSTRACT Granulosa cells from the first (Fl), third (F3) and fifth and sixth (F5–6) preovulatory follicles and the small yellow follicles (SYFs; diameter 6–8 mm) were cultured for 21 h in the absence and presence of murine and human epidermal growth factors, fibroblast growth factor, transforming growth factors α and β-I (TGFα, TGFβ), platelet-derived growth factor and insulin-like growth factor-I at concentrations of 0·1–100 ng/ml. Plasminogen activator (PA) activities in the cell (PAc) and in the medium (PAm) were measured by fibrinolysis and fibrin overlay methods. Basal PAc and PAm activities were highest in cell cultures from the less mature follicles (F5–6 and SYF) and decreased as the follicles matured (F3>F1). PAc activity was greater than PAm activity, irrespective of the stage of follicular development. All growth factors examined at the 100 ng/ml level were effective in increasing PAc and PAm activities in cultures of granulosa cells from Fl follicles. However, only TGFα was able to increase PA activities at lower concentrations. The stimulation of the PA activities of granulosa cells from F3 follicles was inconsistent. None of the growth factors significantly increased PA activities in granulosa cells from F5–6 follicles and SYFs, as determined by fibrinolysis. The major PAc and PAm species (characterized by fibrin overlay) had a molecular mass of about 35 kDa, which is characteristic of the urokinase type. Both assay methods detected a stimulatory effect of the growth factors on PA activities in the granulosa cells from Fl follicles. However, an increase in PA activities in cells from F3 and F5–6 follicles and SYFs was indicated only after fibrin overlay analysis. Tritiated thymidine was incorporated into the DNA of granulosa cells at all stages of follicular development and was enhanced by all growth factors, although TGFα and TGFβ were the most effective and had a ranked order of activity: F3, F5–6>F1, SYF. The present findings show that, of the growth factors examined, TGFα may be an effective regulator of PA activity in avian granulosa cells during follicular development, in addition to its observed mitogenic action.

scholarly journals Differential expression and regulation of prostaglandin E synthases in the mouse ovary during sexual maturation and luteal development

2006 ◽  
Vol 189 (1) ◽  
pp. 89-101 ◽  
Author(s):  
Tong Sun ◽  
Wen-Bo Deng ◽  
Hong-Lu Diao ◽  
Hua Ni ◽  
Yu-Yan Bai ◽  
...  
Keyword(s):  
Corpus Luteum ◽  
Chorionic Gonadotropin ◽  
Granulosa Cells ◽  
Sexual Maturation ◽  
Mrna Level ◽  
Follicular Development ◽  
Prostaglandin E ◽  
Female Reproduction ◽  
Mouse Ovary ◽  
Equine Chorionic Gonadotropin

Prostaglandin (PGE) 2 is the most common prostanoid and plays an important role in female reproduction. The aim of this study was to examine the expression and regulation of microsomal (m) PGE synthase (PGES)-1 and cytosolic (c) PGES in the mouse ovary during sexual maturation, gonadotropin treatment and luteal development by in situ hybridization and immunohistochemistry. Both mPGES-1 mRNA signals and immunostaining were localized in the granulosa cells, but not in the thecal cells and oocytes. cPGES mRNA signals were localized in both granulosa cells and oocytes, whereas cPGES immunostaining was exclusively localized in the oocytes. In our superovulated model of immature mice, there was a basal level of mPGES-1 mRNA signals in the granulosa cells at 48 h after equine chorionic gonadotropin (eCG) treatment. mPGES-1 mRNA level was induced by human chorionic gonadotropin (hCG) treatment for 0.5 h, whereas mPGES-1 immunostaining was slightly induced at 0.5 h after hCG treatment and reached a maximal level at 3 h after hCG treatment. eCG treatment had no obvious effects on either cPGES mRNA signals or immunostaining. A strong level of cPGES immunostaining was present in both unstimulated and eCG-treated groups. Both mPGES-1 mRNA signals and immunostaining were highly detected in the corpus luteum 2 days post-hCG injection and declined from days 3 to 7 post-hCG injection. cPGES immunostaining was at a basal level or not detectable from days 1 to 7 after hCG injection and was highly expressed in the corpus luteum from days 9 to 15 post-hCG injection. PGE2 biosynthesized through the mPGES-1 pathway may be important for follicular development, ovulation and luteal formation.

scholarly journals MicroRNA Mediating Networks in Granulosa Cells Associated with Ovarian Follicular Development

2017 ◽  
Vol 2017 ◽  
pp. 1-18 ◽  
Author(s):  
Baoyun Zhang ◽  
Long Chen ◽  
Guangde Feng ◽  
Wei Xiang ◽  
Ke Zhang ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Granulosa Cells ◽  
Messenger Rna ◽  
Expression Patterns ◽  
Follicular Development ◽  
Functional Networks ◽  
Signal Pathways ◽  
Differentially Expressed Mirnas ◽  
Selection Of

Ovaries, which provide a place for follicular development and oocyte maturation, are important organs in female mammals. Follicular development is complicated physiological progress mediated by various regulatory factors including microRNAs (miRNAs). To demonstrate the role of miRNAs in follicular development, this study analyzed the expression patterns of miRNAs in granulosa cells through investigating three previous datasets generated by Illumina miRNA deep sequencing. Furthermore, via bioinformatic analyses, we dissected the associated functional networks of the observed significant miRNAs, in terms of interacting with signal pathways and transcription factors. During the growth and selection of dominant follicles, 15 dysregulated miRNAs and 139 associated pathways were screened out. In comparison of different styles of follicles, 7 commonly abundant miRNAs and 195 pathways, as well as 10 differentially expressed miRNAs and 117 pathways in dominant follicles in comparison with subordinate follicles, were collected. Furthermore, SMAD2 was identified as a hub factor in regulating follicular development. The regulation of miR-26a/b onsmad2messenger RNA has been further testified by real time PCR. In conclusion, we established functional networks which play critical roles in follicular development including pivotal miRNAs, pathways, and transcription factors, which contributed to the further investigation about miRNAs associated with mammalian follicular development.

A STUDY OF BLASTOCYSTS DURING DELAY AND SUBSEQUENT IMPLANTATION IN LACTATING MICE

1968 ◽  
Vol 42 (3) ◽  
pp. 453-463 ◽  
Author(s):  
ANNE McLAREN
Keyword(s):  
Dna Synthesis ◽  
Thymidine Incorporation ◽  
Tritiated Thymidine ◽  
Uterine Horn ◽  
Uterine Epithelium ◽  
Serial Sections ◽  
Uterine Lumen ◽  
Fresh State

SUMMARY Blastocysts were studied on the 5th and 8th day of pregnancy in lactating mice, in the fresh state, flushed from the uterus, in squash preparations and in serial sections. At the earlier period some mitosis was observed. Tritiated thymidine incorporation studies gave some evidence of DNA synthesis on the 5th and 6th days of pregnancy. By the 8th day the blastocysts were longer, contained more cells, and mitosis had ceased. They were located at the anti-mesometrial end of the uterine lumen, closely apposed to the uterine epithelium, and with their long axes parallel to the long axis of the uterine horn. Implantation could be induced, either by the removal of the litter, or by the injection of an appropriate dose of oestrogen on the 5th or 7th (but not the 4th) day of pregnancy. Both treatments were followed by the appearance of W-bodies in the neighbourhood of the blastocysts, the disappearance of the shed zonae, and the appearance of Pontamine Blue reactivity, oedema of the uterine stroma and formation of the primary decidual zone, in that order.

scholarly journals AKT is involved in granulosa cell autophagy regulation via mTOR signaling during rat follicular development and atresia

Reproduction ◽  
2014 ◽  
Vol 147 (1) ◽  
pp. 73-80 ◽  
Author(s):  
JongYeob Choi ◽  
MinWha Jo ◽  
EunYoung Lee ◽  
DooSeok Choi
Keyword(s):  
Cell Death ◽  
Granulosa Cell ◽  
Granulosa Cells ◽  
Apoptotic Cell ◽  
Follicular Development ◽  
Negative Regulator ◽  
Metabolic Clearance ◽  
Akt Inhibitor ◽  
Western Blots

In this study, we examined whether granulosa cell autophagy during follicular development and atresia was regulated by the class I phosphoinositide-3 kinase/protein kinase B (AKT) pathway, which is known to control the activity of mammalian target of rapamycin (mTOR), a major negative regulator of autophagy. Ovaries and granulosa cells were obtained using an established gonadotropin-primed immature rat model that induces follicular development and atresia. Autophagy was evaluated by measuring the expression level of microtubule-associated protein light chain 3-II (LC3-II) using western blots and immunohistochemistry. The activity of AKT and mTOR was also examined by observing the phosphorylation of AKT and ribosomal protein S6 kinase (S6K) respectively. After gonadotropin injection, LC3-II expression was suppressed and phosphorylation of AKT and S6K increased in rat granulosa cells. By contrast, gonadotropin withdrawal by metabolic clearance promoted LC3-II expression and decreased phosphorylation of AKT and S6K. In addition,in-vitroFSH treatment of rat granulosa cells also indicated inhibition of LC3-II expression accompanied by a marked increase in phosphorylation of AKT and S6K. Inhibition of AKT phosphorylation using AKT inhibitor VIII suppressed FSH-mediated phosphorylation of S6K, followed by an increase in LC3-II expression. Furthermore, co-treatment with FSH and AKT inhibitor increased the levels of apoptosis and cell death of granulosa cells compared with the single treatment with FSH. Taken together, our findings indicated that AKT-mediated activation of mTOR suppresses granulosa cell autophagy during follicular development and is involved in the regulation of apoptotic cell death.

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