Dataset Information

Changes of miRNA expression profiling in GT1-7 cells after treated with dexamethasone(DEX)

ABSTRACT: Stress is defined as a systemic nonspecific adaptive response to the stimulations from internal and external environment or psychological factors through coordinating series of complex systems, like nervous system, immune system and respiratory system. While it is well known that stress have profound negative effects on reproductive function, the mechanism of reproductive dysfunction is still far from perfect. Kisspeptin, which are known for regulating GnRH synthesis and secretion, affect the initiation of puberty and reproduction, have been shown to express glucocorticoid receptors(GR),and both acute and chronic stress significantly inhibit Kisspeptin expression. So, this study speculated that Kisspeptin might participate in the process of reproductive regulation during stress. However, when constructed stress models of GT1-7 cells(Derived from mouse hypothalamic neurons, could not only transcribe endogenous GnRH mRNA and release GnRH when depolarization, but also express Kisspeptin, G protein-coupled receptors) with DEX and tried to analyze the specific mechanism of stress inhibited Kisspeptin expression, opposite results(Dex improved Kisspeptin expression of GT1-7 cells) were obtained. Based on the neuroimmune system hypothesis of stress, this study speculated that the direct reason of stress inhibited Kisspeptin expression might be stress-induced inflammatory response rather than stress-elevated GC. Even so, the transcription of Il-1β decreased in BV-2 cells(Derived from mouse Microglia) after treated with DEX. Due to the proximity of Kisspeptin neurons and Microgila in arcuate nucleus(ARC) of hypothalamus, it is speculated that there maight be interaction between them. So, this study co-cultured GT1-7 cells and BV-2 cells, and found transcription of Kiss1 decreased and Il-1β increased. Decreased Kisspeptin expression due to inflammation of BV-2 cells and inflammation of BV-2 cells must due to changes of certain substances or molecules in GT1-7 cells after treated with DEX. To further explore the possible mechanism, this study used the highly sensitive miRNAs array to screen miRNAs which not only changed significantly in GT1-7 cells after treated with DEX but also associated with inflammation. Finally, 10 miRNAs obtained and further confirmed mmu-miR-146a-5P could inhibited the LPS-induced BV-2 cells inflammation, M1 polarization and apoptosis.

ORGANISM(S): Mus musculus Rattus norvegicus Homo sapiens

PROVIDER: GSE230086 | GEO | 2025/12/31

REPOSITORIES: GEO

ACCESS DATA

Dataset's files

Source:

			Action	DRS
		Other

Items per page:

1 - 1 of 1

Similar Datasets

Project description:Fertility critically depends on the gonadotropin-releasing hormone (GnRH) pulse generator, a neural construct comprised of hypothalamic neurons coexpressing kisspeptin, neurokoinin-B and dynorphin. Here, using mathematical modeling and in vivo optogenetics we reveal for the first time how this neural construct initiates and sustains the appropriate ultradian frequency essential for reproduction. Prompted by mathematical modeling, we show experimentally using female estrous mice that robust pulsatile release of luteinizing hormone, a proxy for GnRH, emerges abruptly as we increase the basal activity of the neuronal network using continuous low-frequency optogenetic stimulation. Further increase in basal activity markedly increases pulse frequency and eventually leads to pulse termination. Additional model predictions that pulsatile dynamics emerge from nonlinear positive and negative feedback interactions mediated through neurokinin-B and dynorphin signaling respectively are confirmed neuropharmacologically. Our results shed light on the long-elusive GnRH pulse generator offering new horizons for reproductive health and wellbeing.SIGNIFICANCE STATEMENT The gonadotropin-releasing hormone (GnRH) pulse generator controls the pulsatile secretion of the gonadotropic hormones LH and FSH and is critical for fertility. The hypothalamic arcuate kisspeptin neurons are thought to represent the GnRH pulse generator, since their oscillatory activity is coincident with LH pulses in the blood; a proxy for GnRH pulses. However, the mechanisms underlying GnRH pulse generation remain elusive. We developed a mathematical model of the kisspeptin neuronal network and confirmed its predictions experimentally, showing how LH secretion is frequency-modulated as we increase the basal activity of the arcuate kisspeptin neurons in vivo using continuous optogenetic stimulation. Our model provides a quantitative framework for understanding the reproductive neuroendocrine system and opens new horizons for fertility regulation Model is encoded by Johannes and submitted to BioModels by Ahmad Zyoud.

Project description:The kiss system is known to regulate puberty and to be involved in the onset of reproduction, acting upstream of the Brain-Pituitary-Gonad axis in gonadotrophin releasing hormone neurons and stimulating secretion of GnRH. Kisspeptin has been proven to be a novel therapeutic for fertility disorders in humans. In fish, positive results have also observed the prospective of this hormone to induce gamete development. During the last two decades, support has been obtained describing the key role of microRNAs (miRNA) in regulating gene expression at the post-transcriptional level and they have been recommended as relevant biomarkers in reproductive studies. Senegalese sole is a marine flatfish species with high importance in the European aquaculture industry, due to its good growth rate and high market value. However, this species presents a bottleneck in terms of reproduction, which does not allow to close the cycle in captive conditions. Hence, this study aimed to observe the effect of a novel hormonal treatment using kisspeptin in miRNA expression related to reproductive performance in cultured Senegalese sole breeders. For this purpose, Senegalese sole F1 broodstock were held at CCMAR facilities in 4 tanks of 3m3 (n = 12/per tank, sex ratio 1:1) under natural photoperiod and temperature conditions. During the reproductive season, the animals from each 2 of the tanks were treated either with SSkiss2_v2 decapeptide in a single intramuscular injection, or with a PBS injection for control groups. After the treatment (2 and 4 days), all fish were anesthetized and sampled for blood collection. Afterwards, plasma was obtained to determine several parameters related to reproductive physiology (Fsh, Lh and sexual esteroids (T, 11-KT and E2)) and miRNAs regulation. After observing the high implication of this treatment in the gonadotrofins, above all in Fsh and sexual esteroids, above all in T, the RNA was extracted from blood plasma using miRNeasy Serum/Plasma Kit (Qiagen, Germany) and libraries were prepared using NEXTflex Small RNA-Seq kit v3 (Bioo Scientific, USA). Small RNA sequencing (sRNA-seq) (NextSeq 500) was performed on sole treated with kisspeptin (n = 16) and non-treated control fish (n = 14). A total of 1,467 unique miRNAs were expressed in Senegalese sole plasma. A multidimensional scaling (MDS) plot showed no clear clustering of the samples with the kisspeptin treatment. However, the differential expression analysis revealed one miRNA, miR-1-3p, to be highly up-regulated in both males and females treated with kisspeptin compared to control sole. This miRNA is linked with myogenesis and angiogenesis, and skeletal muscle development in fish. Three isomirs (let 7e, miR-199a-3p, miR-100-5p) were differentially expressed between females treated and control fish. MicroRNA let-7e is an important regulator of endothelial function and inflammation, related to testosterone pathway release; miR-199a-3p plays an important role in inflammation and women fertility and miR-100-5p might be implicated in the regulation of cell growth and oocytes maturation. In conclusion, kisspeptin treatment may affect gonad maturation in females and muscle growth in males. Our data support the possibility of using circulating miRNAs as less invasive physiological biomarkers in fish reproduction.

Dataset Information

Changes of miRNA expression profiling in GT1-7 cells after treated with dexamethasone(DEX)

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets