scholarly journals Prolactin independent rescue of mouse corpus luteum life span: identification of prolactin and luteinizing hormone target genes

2009 ◽  
Vol 297 (3) ◽  
pp. E676-E684 ◽  
Author(s):  
Anne Bachelot ◽  
Julie Beaufaron ◽  
Nathalie Servel ◽  
Cécile Kedzia ◽  
Philippe Monget ◽  
...  
Keyword(s):  
Luteinizing Hormone ◽  
Corpus Luteum ◽  
Molecular Mechanisms ◽  
Target Genes ◽  
Ovarian Function ◽  
De Novo ◽  
Embryo Implantation ◽  
Receptor Expression ◽  
Corpora Lutea ◽  
Mrna Expression Profiling

The corpus luteum (CL) plays a central role in the maintenance of pregnancy in rodents, mainly by secreting progesterone. Female mice lacking prolactin (PRL) receptor (R) are sterile due to a failure of embryo implantation, which is a consequence of decreased luteinizing hormone (LH) receptor expression in the CL and inadequate levels of progesterone. We attempted to treat PRLR−/− females with human chorionic gonadotropin (hCG) and showed a de novo expression of LHR mRNA in the corpora lutea. Binding analysis confirmed that the LHR in hCG-treated PRLR−/− animals was functional. This was accompanied with increased expression of steroidogenic enzymes involved in progesterone synthesis. Despite these effects, no embryo implantation was observed because of high expression of 20α-hydroxysteroid dehydrogenase. To better appreciate the molecular mechanisms underlying maintenance of the CL, a series of mRNA expression-profiling experiments was performed on isolated corpora lutea of PRLR−/− and hCG-treated PRLR−/− mice. This approach revealed several novel candidate genes with potentially pivotal roles in ovarian function, among them, p27, VE-cadherin, Pten, and sFRP-4, a member of the Wnt/frizzled family. This study showed the differential role of PRL and LH in CL function and identified new targets of these hormones in luteal cells.

scholarly journals Examination of the Transcriptional Response to LaMIR166a Overexpression in Larix kaempferi (Lamb.) Carr

Biology ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 576
Author(s):  
Yanru Fan ◽  
Wanfeng Li ◽  
Zhexin Li ◽  
Shaofei Dang ◽  
Suying Han ◽  
...  
Keyword(s):  
Cell Lines ◽  
Molecular Mechanisms ◽  
Target Genes ◽  
De Novo ◽  
Pearson Correlation ◽  
Transcriptional Response ◽  
Correlation Coefficients ◽  
Embryonic Cell ◽  
Larix Kaempferi ◽  
Transgenic Cell

The study of somatic embryogenesis can provide insight into early plant development. We previously obtained LaMIR166a-overexpressing embryonic cell lines of Larix kaempferi (Lamb.) Carr. To further elucidate the molecular mechanisms associated with miR166 in this species, the transcriptional profiles of wild-type (WT) and three LaMIR166a-overexpressing transgenic cell lines were subjected to RNA sequencing using the Illumina NovaSeq 6000 system. In total, 203,256 unigenes were generated using Trinity de novo assembly, and 2467 differentially expressed genes were obtained by comparing transgenic and WT lines. In addition, we analyzed the cleaved degree of LaMIR166a target genes LaHDZ31–34 in different transgenic cell lines by detecting the expression pattern of LaHdZ31–34, and their cleaved degree in transgenic cell lines was higher than that in WT. The downstream genes of LaHDZ31–34 were identified using Pearson correlation coefficients. Yeast one-hybrid and dual-luciferase report assays revealed that the transcription factors LaHDZ31–34 could bind to the promoters of LaPAP, LaPP1, LaZFP5, and LaPHO1. This is the first report of gene expression changes caused by LaMIR166a overexpression in Japanese larch. These findings lay a foundation for future studies on the regulatory mechanism of miR166.

IN-VITRO STEROIDOGENESIS OF NEWLY FORMED CORPORA LUTEA AND THE NON-LUTEAL OVARY IN THE RAT, RABBIT, HAMSTER AND GUINEA-PIG

1980 ◽  
Vol 84 (1) ◽  
pp. 101-108 ◽  
Author(s):  
P. F. TERRANOVA ◽  
S. K. SAIDAPUR ◽  
G. S. GREENWALD
Keyword(s):  
Guinea Pig ◽  
Corpus Luteum ◽  
Ovarian Function ◽  
Serum Testosterone ◽  
The Other ◽  
Corpora Lutea ◽  
Serum Progesterone ◽  
Antral Follicles ◽  
Baseline Levels

The steroidogenic abilities of the newly formed corpus luteum (8–10 h after ovulation) and the non-luteal ovary were compared in the guinea-pig, hamster, rabbit and rat using an invitro incubation technique. Histologically, newly formed rat corpora lutea (CL) were highly luteinized whereas the CL of the rabbit and guinea-pig were only partially luteinized. The CL of the hamster showed the least amount of luteinization. Serum progesterone was highest in the rat (18 ± 3 (s.e.m.) ng/ml). In the hamster, it was about 8 ng/ml, whereas in the rabbit and guinea-pig it was about 1 ng/ml. Serum androstenedione ranged between 0·5 and 1 ng/ml. Serum testosterone was lowest in the hamster (60 pg/ml) and highest in the rabbit (470 pg/ml), whereas in the rat and guinea-pig, testosterone levels were similar (about 240 pg/ml). Serum oestrogens were at baseline levels in all species. The CL of the rat exhibited considerably greater steroidogenic ability than the CL of the other species, producing 70 ± 6 ng progesterone/mg per h, 215 ± 14 pg androstenedione/mg per h, 49 ± 3 pg testosterone/mg per h, 3 pg oestrone/mg per h and 1 pg oestradiol/mg per h. Rabbit CL produced only progesterone (7 ± 2 ng/mg per h). Newly formed hamster CL produced none of the above steroids. In general, the ability of the CL to produce progesterone in vitro correlated with the degree of luteinization found by histological observation. Guinea-pig CL were embedded deeply in the ovary and could not be obtained without damage. Consequently, a portion of the ovary containing a corpus luteum was incubated. There was no difference in the steroid production by this portion of the ovary compared with the non-luteal ovary. The non-luteal ovary of the rat produced the highest amount of progesterone (10 ± 2 ng/mg per h). The guinea-pig non-luteal ovary produced about 5 ± 2 ng progesterone/mg per h, whereas the non-luteal ovary of the rabbit did not produce any. On the other hand, the hamster non-luteal ovary lost progesterone. Non-luteal ovaries from all species produced androgens. The non-luteal ovary of the guinea-pig contained especially large numbers of atretic antral follicles. The guinea-pig non-luteal ovary produced extremely large amounts of androstenedione (1110 ± 210 pg/mg per h) and testosterone (606 ± 154 pg/mg per h) compared with the amounts produced by the non-luteal ovary of the rat, hamster and rabbit. In the non-luteal ovary, interstitium and atretic antral follicles are the probable source of androgens. Oestrogen production by the non-luteal ovary was at baseline levels in the four species studied correlating with the absence of healthy antral follicles. The results indicate the extreme species differences that exist in ovarian function in the early postovulatory period.

SUPERFUSION IN VITRO IN THE STUDY OF OVARIAN STEROIDOGENESIS

1975 ◽  
Vol 64 (1) ◽  
pp. 163-173 ◽  
Author(s):  
JOHN WATSON ◽  
J. T. S. LEASK
Keyword(s):  
Luteinizing Hormone ◽  
Corpus Luteum ◽  
Transient Increase ◽  
Corpora Lutea ◽  
Ovarian Steroidogenesis ◽  
Steroid Secretion ◽  
Regulatory Factors ◽  
Prolonged Response

SUMMARY A method for the continuous superfusion of porcine corpus luteum tissue is described which readily allows both the introduction of regulatory factors to the incubating tissue, and sampling of the tissue. Oestrogen (principally oestradiol) and progestin (principally progesterone) can be measured for up to 24 h in the superfusate from corpora lutea of all ages, and the secretion of both steroids is stimulated by the addition of luteinizing hormone. The pattern of response of both steroids to a pulse of gonadotrophin was similar in that a rapid transient increase in secretion occurred followed some time later by a secondary and more prolonged response. A second pulse of gonadotrophin introduced 6 h after the first also stimulated steroid secretion, indicating that during superfusion in vitro the porcine corpus luteum does not become refractory to the steroidogenic effect of gonadotrophin.

scholarly journals Transcriptomic Analysis Identifies New Non-Target Site Glyphosate-Resistance Genes in Conyza bonariensis

Plants ◽  
2019 ◽  
Vol 8 (6) ◽  
pp. 157 ◽  
Author(s):  
Cristiano Piasecki ◽  
Yongil Yang ◽  
Daiane P. Benemann ◽  
Frederico S. Kremer ◽  
Vanessa Galli ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Target Genes ◽  
De Novo ◽  
Average Length ◽  
Transcriptome Assembly ◽  
Glyphosate Resistance ◽  
Target Site ◽  
Differentially Expressed ◽  
Irreversible Damage ◽  
Conyza Bonariensis

Conyza bonariensis (hairy fleabane) is one of the most problematic and widespread glyphosate-resistant weeds in the world. This highly competitive weed species significantly interferes with crop growth and substantially decreases crop yield. Despite its agricultural importance, the molecular mechanisms of glyphosate resistance are still unknown. The present RNA-Seq study was performed with the goal of identifying differentially expressed candidate transcripts (genes) related to metabolism-based non-target site glyphosate resistance in C. bonariensis. The whole-transcriptome was de novo assembled from glyphosate-resistant and -sensitive biotypes of C. bonariensis from Southern Brazil. The RNA was extracted from untreated and glyphosate-treated plants at several timepoints up to 288 h after treatment in both biotypes. The transcriptome assembly produced 90,124 contigs with an average length of 777 bp and N50 of 1118 bp. In response to glyphosate treatment, differential gene expression analysis was performed on glyphosate-resistant and -sensitive biotypes. A total of 9622 genes were differentially expressed as a response to glyphosate treatment in both biotypes, 4297 (44.6%) being up- and 5325 (55.4%) down-regulated. The resistant biotype presented 1770 up- and 2333 down-regulated genes while the sensitive biotype had 2335 and 2800 up- and down-regulated genes, respectively. Among them, 974 up- and 1290 down-regulated genes were co-expressed in both biotypes. In the present work, we identified 41 new candidate target genes from five families related to herbicide transport and metabolism: 19 ABC transporters, 10 CYP450s, one glutathione S-transferase (GST), five glycosyltransferases (GT), and six genes related to antioxidant enzyme catalase (CAT), peroxidase (POD), and superoxide dismutase (SOD). The candidate genes may participate in metabolic-based glyphosate resistance via oxidation, conjugation, transport, and degradation, plus antioxidation. One or more of these genes might ‘rescue’ resistant plants from irreversible damage after glyphosate treatment. The 41 target genes we report in the present study may inform further functional genomics studies, including gene editing approaches to elucidate glyphosate-resistance mechanisms in C. bonariensis.

Induction of ovulation during anoestrus in two breeds of sheep with multiple injections of LH alone or in combination with FSH

1986 ◽  
Vol 111 (1) ◽  
pp. 181-190 ◽  
Author(s):  
J. M. Wallace ◽  
A. S. McNeilly ◽  
D. T. Baird
Keyword(s):  
Luteinizing Hormone ◽  
Corpus Luteum ◽  
Corpora Lutea ◽  
Preovulatory Follicle ◽  
Treatment Timing ◽  
Multiple Injections ◽  
Induction Of Ovulation ◽  
Preovulatory Follicles ◽  
Identical Manner ◽  
Hormonal Priming

ABSTRACT The gonadotrophic requirements for the induction of ovulation and formation of a viable corpus luteum in two breeds of seasonally anoestrous sheep of differing fecundity was investigated. Welsh Mountain (n = 20) and Damline ewes (n = 19) were given LH or FSH either alone or in combination. Luteinizing hormone was injected i.v. at an increasing frequency for 72 h (one injection every 3 h for 24 h, one every 2 h for 24 h, and one every hour for 24 h) and FSH was injected in an identical manner for the first 36 h of treatment. Exogenous LH alone and in combination with FSH induced a preovulatory LH surge in all 19 ewes and ovulation in 18 out of 19 ewes of both breeds. However exogenous FSH alone was ineffective. The incidence of normal corpus luteum function in ewes induced to ovulate was low and not related to treatment, timing or magnitude of the LH/FSH surge. It is concluded that in both breeds studied (a) it is the infrequency of LH pulses which limits the development of preovulatory follicles during seasonal anoestrus, (b) that the requirement for FSH remains unknown, and (c) that the induction of inadequate corpora lutea during seasonal anoestrus reflects either defects in hormonal priming of the preovulatory follicle and/or inappropriate luteotrophic support after ovulation. J. Endocr. (1986) 111, 181–190

Oxytocin determination in steroid producing tissues and in vitro production in ovarian follicles

1985 ◽  
Vol 109 (4) ◽  
pp. 530-536 ◽  
Author(s):  
D. Schams ◽  
Th. A. M. Kruip ◽  
R. Koll
Keyword(s):  
Corpus Luteum ◽  
Granulosa Cells ◽  
Follicular Fluid ◽  
De Novo ◽  
Prostate Gland ◽  
Seminal Vesicles ◽  
Corpora Lutea ◽  
In Vitro Production ◽  
Wet Weight

Abstract Immunoreactive oxytocin was measured in ovaries (corpus luteum and follicular fluid) and adrenals of cows, and in testes, seminal vesicles, prostate gland and adrenals of bulls. Secretion of oxytocin was further measured after culture of whole follicles, granulosa cells and theca tissue. Concentrations of oxytocin increased in corpora lutea of cycling cattle until mid-luteal phase (447 ± 93 ng/g wet weight) and decreased afterwards. Low concentrations were found in corpora lutea of pregnant animals (6 ± 3 ng/g wet weight). Follicular fluid contains some oxytocin (on average 42–108 pg/ml) but concentrations were significantly higher in the fluid of ovarian cysts (190 pg/ml). After culture of follicles the amount of oxytocin released into the medium increased indicating de novo synthesis. The granulosa cells were the main source of follicular oxytocin. Production increased during luteinization indicating that luteinization is an important step for the production of oxytocin in ovaries. Tissues of testes (65 ± 10 pg/g wet weight) and adrenals from cows (122 ± 39 pg/g wet weight) and bulls (111 ±2 pg/g wet weight) contained oxytocin but at much lower concentrations compared to corpus luteum tissue. About 10 times higher concentrations of oxytocin were measured in the adrenal medulla (717 ± 96 pg/g wet weight) compared to the cortex (72 ± 11 pg/g wet weight). Seminal vesicles and prostate gland contained no measurable amounts of oxytocin (< 5 pg/g wet weight).

scholarly journals The ETS Transcription Factor ERF controls the exit from the naïve pluripotent state

2021 ◽  
Author(s):  
M. Vega-Sendino ◽  
T. Olbrich ◽  
D. Tillo ◽  
A. D. Tran ◽  
C. N. Domingo ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Molecular Mechanisms ◽  
De Novo ◽  
Embryo Implantation ◽  
Ras Proteins ◽  
Transcriptional Program ◽  
Nanog Expression ◽  
Downstream Effectors ◽  
Primed State ◽  
Ets Transcription Factor

The naïve epiblast undergoes a transition to a pluripotent primed state during embryo implantation. Despite the relevance of the FGF pathway during this period, little is known about the downstream effectors regulating this signaling. Here, we examined the molecular mechanisms coordinating the naïve to primed transition by using inducible ESC to genetically eliminate all RAS proteins. We show that differentiated RASKO ESC remain trapped in an intermediate state of pluripotency with naïve-associated features. Elimination of the transcription factor ERF overcomes the developmental blockage of RAS-deficient cells by naïve enhancer decommissioning. Mechanistically, ERF regulates NANOG expression and ensures naïve pluripotency by strengthening naïve transcription factor binding at ESC enhancers. Moreover, ERF negatively regulates the expression of the de novo methyltransferase DNMT3B, which participates in the extinction of the naïve transcriptional program. Collectively, we demonstrated an essential role for ERF controlling the exit from naïve pluripotency during the progression to primed pluripotency.TeaserERF is the MAPK-dependent switch controlling the transition between naïve and primed pluripotency during embryonic development.

scholarly journals Interaction of Mouse Placental Lactogens and Androgens in Regulating Progesterone Release in Cultured Mouse Luteal Cells

Endocrinology ◽  
1997 ◽  
Vol 138 (8) ◽  
pp. 3236-3241 ◽  
Author(s):  
G. Thordarson ◽  
S. Galosy ◽  
G. O. Gudmundsson ◽  
B. Newcomer ◽  
R. Sridaran ◽  
...  
Keyword(s):  
Androgen Receptor ◽  
Corpus Luteum ◽  
De Novo ◽  
Pituitary Hormones ◽  
Pregnant Mouse ◽  
Placental Lactogen ◽  
Luteal Cell ◽  
Corpora Lutea ◽  
Luteal Cells

Abstract Pituitary hormones are essential for the maintenance of the corpus luteum in the pregnant mouse during the first half of gestation. Thereafter, hormones from the placenta take over the luteotropic role of the pituitary hormones. Mouse placental lactogen-I (mPL-I) and mPL-II, two PRL-like hormones produced in the placenta, are probably necessary for the maintenance of the corpus luteum in the latter half of pregnancy. A culture system of luteal cells from pregnant mice was developed to investigate the role of hormones from the placenta that may be important for the function of the corpus luteum. Mice were killed on days 10, 14, and 18 of pregnancy, and the corpora lutea were excised from the ovaries and digested in 0.1% collagenase, 0.002% DNase for 1 h. The resulting luteal cell suspension was plated onto 96-well plates coated with fibronectin (1 × 105 cells/well) and cultured for 1–3 days. Medium was changed daily. The cells were treated with various concentrations and combinations of mPL-I, mPL-II, mouse PRL, androstenedione, dihydrotestosterone, 17β-estradiol (E2), testosterone, hydroxyflutamide, cycloheximide, actinomycin D, and fadrozole to study the effects of these different treatments on progesterone (P4) production. The three lactogens (mPL-I, mPL-II, and mouse PRL) all stimulated the release of P4 from the luteal cells. The potency of the lactogens was similar and did not depend on the stage of pregnancy at which the luteal tissue was obtained. However, the responsiveness of the cells to all hormone-stimulated P4 release was gradually reduced the later in pregnancy the tissue was collected. Androgens also stimulated the release of P4 from the luteal cells, and when administered together, the lactogens and the androgens acted synergistically to stimulate P4 release. The androgens acted directly but not through conversion to E2, as determined by the findings that 1) the effects of the androgens could not be reproduced by E2 administration, 2) nonaromatizable androgen dihydrotestosterone was as effective as aromatizable androgens, and 3) aromatase inhibitor did not prevent the action of the androgens to stimulate the P4 release. The effect of the androgens on the P4 release was rapid, occurring within 15 min of hormone administration. It was not prevented by inhibitors of protein and RNA synthesis, and the intracellular androgen receptor antagonist hydroxyflutamide did not affect the androgen action. Therefore, the androgen effects were not mediated through the intracellular androgen receptor and de novo protein synthesis was not needed for androgen-stimulated P4 release.

OVULATION AND CORPUS LUTEUM FUNCTION IN THE LLAMA (LAMA GLAMA)

1969 ◽  
Vol 45 (4) ◽  
pp. 505-513 ◽  
Author(s):  
B. G. ENGLAND ◽  
W. C. FOOTE ◽  
D. H. MATTHEWS ◽  
ARMANDO G. CARDOZO ◽  
S. RIERA
Keyword(s):  
Luteinizing Hormone ◽  
Corpus Luteum ◽  
Breeding Season ◽  
Luteal Phase ◽  
Maximum Size ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Corpora Lutea ◽  
Lama Glama

SUMMARY Results in 53 llamas (33 mated animals and 20 controls) showed that ovulation is copulation-induced in this species. Ovulation without copulation occasionally occurred during the height of the recognized breeding season in Bolivia. The first mating during the luteal phase (12–24 days after the preceding ovulation) resulted in ovulation in four out of ten llamas. Determination of pituitary luteinizing hormone (LH) content showed the highest level on the day before mating (9·00 μg./mg.) and the lowest level on day 4 (6·25 μg./mg.). LH level on day 8 was significantly higher than on day 4 (7·62 μg./mg.). Corpora lutea (c.l.) were well formed on day 4 after mating (408 mg.), reached a maximum size by day 8 (1920 mg.) and rapidly decreased in size to day 16 (136 mg.). The corpus albicans remained as an entity but decreased in size to 21 mg. on day 120. Similar changes were found in c.l. histology and progesterone content. The combined results indicate that the functional life of the c.l. in a non-pregnant llama is 16 days or less. Treatment with 25 i.u. human chorionic gonadotrophin was sufficient to cause ovulation in 50% of the animals treated. A large (150 mg.) dose of norethandrolone did not cause morphological regression of the c.l. when measured 5 days after treatment. Treatment with 5 mg. daily for 14 days caused regression of c.l. as compared with untreated controls and animals treated with oestradiol valerate.

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