Stem cell factor promotes in vitro ovarian follicle development in the domestic cat by upregulating c-kit mRNA expression and stimulating the phosphatidylinositol 3-kinase/AKT pathway

2017 ◽  
Vol 29 (7) ◽  
pp. 1356 ◽  
Author(s):  
Paweena Thuwanut ◽  
Pierre Comizzoli ◽  
David E. Wildt ◽  
Carol L. Keefer ◽  
Nucharin Songsasen
Keyword(s):  
Stem Cell ◽  
Mrna Expression ◽  
Stem Cell Factor ◽  
Ovarian Follicle ◽  
Domestic Cat ◽  
Follicle Development ◽  
Phosphatidylinositol 3 Kinase ◽  
Akt Phosphorylation ◽  
Phosphatidylinositol 3

In the present study we examined the effects of stem cell factor (SCF; 50 vs 100 ng mL–1) alone or in combination with epidermal growth factor (EGF; 100 ng mL–1) on: (1) the in vitro viability and growth of cat follicles within ovarian cortices; (2) phosphatidylinositol 3-kinase (PI3K)/AKT and mitogen-activated protein kinase (MAPK) phosphorylation; and (3) c-kit and FSH receptor (FSHr) mRNA expression. At 100 ng mL–1, SCF increased (P ≤ 0.05) the percentage and size of secondary follicles after 14 days of in vitro culture and sustained AKT phosphorylation after 3 days incubation. EGF suppressed this beneficial effect and reduced (P ≤ 0.05) the percentage of structurally normal follicles and FSHr expression when combined with 100 ng mL–1 SCF. Expression of c-kit mRNA was higher (P ≤ 0.05) in the presence of 100 ng mL–1 SCF compared with fresh follicles and cohorts cultured under other conditions. A c-kit inhibitor suppressed follicle growth and reduced AKT phosphorylation. Collectively, the results demonstrate that SCF promotes cat follicle development by upregulating c-kit mRNA expression and AKT phosphorylation. EGF suppresses the stimulating effect of SCF, leading to downregulation of FSHr expression.

Stem cell factor induces phosphatidylinositol 3′-kinase–dependent Lyn/Tec/Dok-1 complex formation in hematopoietic cells

Blood ◽  
2000 ◽  
Vol 96 (10) ◽  
pp. 3406-3413 ◽  
Author(s):  
Thamar B. van Dijk ◽  
Emile van den Akker ◽  
Martine Parren-van Amelsvoort ◽  
Hiroyuki Mano ◽  
Bob Löwenberg ◽  
...  
Keyword(s):  
Stem Cell ◽  
Tyrosine Kinase ◽  
Stem Cell Factor ◽  
Hematopoietic Cells ◽  
Sh2 Domain ◽  
Stable Complex ◽  
Phosphatidylinositol 3 Kinase ◽  
Sh2 Domains ◽  
And Migration ◽  
Phosphatidylinositol 3

Stem cell factor (SCF) has an important role in the proliferation, differentiation, survival, and migration of hematopoietic cells. SCF exerts its effects by binding to cKit, a receptor with intrinsic tyrosine kinase activity. Activation of phosphatidylinositol 3′-kinase (PI3-K) by cKit was previously shown to contribute to many SCF-induced cellular responses. Therefore, PI3-K-dependent signaling pathways activated by SCF were investigated. The PI3-K-dependent activation and phosphorylation of the tyrosine kinase Tec and the adapter molecule p62Dok-1 are reported. The study shows that Tec and Dok-1 form a stable complex with Lyn and 2 unidentified phosphoproteins of 56 and 140 kd. Both the Tec homology and the SH2 domain of Tec were identified as being required for the interaction with Dok-1, whereas 2 domains in Dok-1 appeared to mediate the association with Tec. In addition, Tec and Lyn were shown to phosphorylate Dok-1, whereas phosphorylated Dok-1 was demonstrated to bind to the SH2 domains of several signaling molecules activated by SCF, including Abl, CrkL, SHIP, and PLCγ-1, but not those of Vav and Shc. These findings suggest that p62Dok-1 may function as an important scaffold molecule in cKit-mediated signaling.

Kit/stem cell factor receptor-induced activation of phosphatidylinositol 3′-kinase is essential for male fertility

10.1038/72814 ◽  
2000 ◽  
Vol 24 (2) ◽  
pp. 157-162 ◽  
Author(s):  
Peter Blume-Jensen ◽  
Guoqiang Jiang ◽  
Robert Hyman ◽  
Kuo-Fen Lee ◽  
Stephen O'Gorman ◽  
...  
Keyword(s):  
Stem Cell ◽  
Male Fertility ◽  
Stem Cell Factor ◽  
Phosphatidylinositol 3 Kinase ◽  
Phosphatidylinositol 3

Activation of elastin transcription by transforming growth factor-β in human lung fibroblasts

2007 ◽  
Vol 292 (4) ◽  
pp. L944-L952 ◽  
Author(s):  
Ping-Ping Kuang ◽  
Xiao-Hui Zhang ◽  
Celeste B. Rich ◽  
Judith A. Foster ◽  
Mangalalaxmy Subramanian ◽  
...  
Keyword(s):  
Growth Factor ◽  
Mrna Expression ◽  
Transforming Growth Factor ◽  
Transforming Growth Factor Β ◽  
Lung Fibroblasts ◽  
Mrna Levels ◽  
Phosphatidylinositol 3 Kinase ◽  
Akt Phosphorylation ◽  
Mapk Inhibitor ◽  
Phosphatidylinositol 3

Elastin synthesis is essential for lung development and postnatal maturation as well as for repair following injury. Using human embryonic lung fibroblasts that express undetectable levels of elastin as assessed by Northern analyses, we found that treatment with exogenous transforming growth factor-β (TGF-β) induced rapid and transient increases in levels of elastin heterogeneous nuclear RNA (hnRNA) followed by increases of elastin mRNA and protein expression. In fibroblasts derived from transgenic mice, TGF-β induced increases in the expression of a human elastin gene promoter fragment driving a chloramphenicol acetyl transferase reporter gene. The induction of elastin hnRNA and mRNA expression by TGF-β was abolished by pretreatments with TGF-β receptor I inhibitor, global transcription inhibitor actinomycin D, and partially blocked by addition of protein synthesis inhibitor cycloheximide, but was not affected by the p44/42 MAPK inhibitor U0126. Pretreatment with the p38 MAPK inhibitor SB-203580 also partially attenuated the levels of TGF-β-induced elastin mRNA but not its hnRNA. Western analysis indicated that TGF-β stimulated Akt phosphorylation. Inhibition of phosphatidylinositol 3-kinase and Akt phosphorylation by LY-294002 abolished TGF-β-induced increases in elastin hnRNA and mRNA expression. Treatment of lung fibroblasts with interleukin-1β or the histone deacetylase inhibitor trichostatin A inhibited TGF-β-induced elastin mRNA and hnRNA expression by a mechanism that involved inhibition of Akt phosphorylation. Downregulation of Akt2 but not Akt1 expression employing small interfering RNA duplexes blocked TGF-β-induced increases of elastin hnRNA and mRNA levels. Together, our results demonstrated that TGF-β activates elastin transcription that is dependent on phosphatidylinositol 3-kinase/Akt activity.

scholarly journals Effects of Stem Cell Factor/c-Kit Signaling on In Vitro Maturation of Porcine Oocytes and Subsequent Developmental Competence After Fertilization

2021 ◽  
Vol 8 ◽  
Author(s):  
Eunhye Kim ◽  
Lian Cai ◽  
Sang-Hwan Hyun
Keyword(s):  
Stem Cell ◽  
Mrna Expression ◽  
Stem Cell Factor ◽  
Polar Body ◽  
Cumulus Cells ◽  
Developmental Competence ◽  
Control Group ◽  
Secreted Factors ◽  
Polar Body Extrusion

Stem cell factor (SCF), also known as c-Kit ligand, plays an important role in the proliferation of primordial germ cells and the survival of oocytes during follicular development. The aim of this study was to investigate the effect of SCF/c-Kit signaling on in vitro maturation (IVM) of porcine oocytes by analyzing nuclear and cytoplasmic maturation, oocyte size, cumulus cell expansion, and developmental competence to the blastocyst stage. Moreover, mRNA expression patterns of porcine cumulus cells and oocytes were evaluated using qRT-PCR. Following 42 h of IVM, 10 and 50 ng/mL SCF-treated groups exhibited significantly (P < 0.05) increased polar body extrusion rates and intracellular glutathione levels compared with the control group. The cumulus expansion index significantly (P < 0.05) increased in all SCF-treated groups compared with the control samples. mRNA levels of the proapoptotic gene Bax and apoptosis-related cysteine peptidase Caspase3 were lower in SCF-treated cumulus cells than in the control group. Notably, the diameter of oocytes after IVM, the mRNA expression of well-known oocyte-secreted factors (GDF9 and BMP15), and an oocyte-specific protein essential for ovulation and oocyte health (YBX2) were significantly (P < 0.05) higher in SCF-treated than in non-treated oocytes. Inhibition of c-Kit during porcine IVM using ACK2, an antagonistic blocker of c-Kit, significantly (P < 0.05) decreased the polar body extrusion rate compared with the control, as well as blastocyst formation rate compared with the 10 ng/mL SCF-treated group. In conclusion, the effect of SCF/c-Kit-mediated signaling during porcine IVM could be ascribed to the reduced expression of apoptosis-related genes and higher expression of oocyte-specific/secreted factors.

Stem cell factor induces phosphatidylinositol 3′-kinase–dependent Lyn/Tec/Dok-1 complex formation in hematopoietic cells

Blood ◽  
2000 ◽  
Vol 96 (10) ◽  
pp. 3406-3413 ◽  
Author(s):  
Thamar B. van Dijk ◽  
Emile van den Akker ◽  
Martine Parren-van Amelsvoort ◽  
Hiroyuki Mano ◽  
Bob Löwenberg ◽  
...  
Keyword(s):  
Stem Cell ◽  
Tyrosine Kinase ◽  
Stem Cell Factor ◽  
Hematopoietic Cells ◽  
Sh2 Domain ◽  
Stable Complex ◽  
Phosphatidylinositol 3 Kinase ◽  
Sh2 Domains ◽  
And Migration ◽  
Phosphatidylinositol 3

Abstract Stem cell factor (SCF) has an important role in the proliferation, differentiation, survival, and migration of hematopoietic cells. SCF exerts its effects by binding to cKit, a receptor with intrinsic tyrosine kinase activity. Activation of phosphatidylinositol 3′-kinase (PI3-K) by cKit was previously shown to contribute to many SCF-induced cellular responses. Therefore, PI3-K-dependent signaling pathways activated by SCF were investigated. The PI3-K-dependent activation and phosphorylation of the tyrosine kinase Tec and the adapter molecule p62Dok-1 are reported. The study shows that Tec and Dok-1 form a stable complex with Lyn and 2 unidentified phosphoproteins of 56 and 140 kd. Both the Tec homology and the SH2 domain of Tec were identified as being required for the interaction with Dok-1, whereas 2 domains in Dok-1 appeared to mediate the association with Tec. In addition, Tec and Lyn were shown to phosphorylate Dok-1, whereas phosphorylated Dok-1 was demonstrated to bind to the SH2 domains of several signaling molecules activated by SCF, including Abl, CrkL, SHIP, and PLCγ-1, but not those of Vav and Shc. These findings suggest that p62Dok-1 may function as an important scaffold molecule in cKit-mediated signaling.

scholarly journals RFRP3 influences basal lamina degradation, cellular death, and progesterone secretion in cultured preantral ovarian follicles from the domestic cat

PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e7540 ◽  
Author(s):  
Kathryn Wilsterman ◽  
George E. Bentley ◽  
Pierre Comizzoli
Keyword(s):  
Basal Lamina ◽  
Captive Breeding ◽  
Direct Action ◽  
Ovarian Follicle ◽  
Ovarian Follicles ◽  
Domestic Cat ◽  
Follicle Development ◽  
Carnivore Species ◽  
Percentage Survival

The hypothalamic neuropeptide RFRP3 can suppress hypothalamic GnRH neuron activation and inhibit gonadotropin release from the anterior pituitary. RFRP3 is also produced locally in the ovary and can inhibit steroidogenesis and follicle development in many vertebrates. However, almost nothing is known about the presence and regulatory action of RFRP3 in gonads of any carnivore species. Such knowledge is important for developing captive breeding programs for endangered carnivores and for inhibiting reproduction in feral species. Using the domestic cat as a model, our objectives were to (1) demonstrate the expression of feline RFRP3 (fRFRP3) and its receptor in the cat ovary and (2) assess the influence of fRFRP3 on ovarian follicle integrity, survival, and steroidogenesis in vitro. We first confirmed that fRFRP3 and its receptors (NPFFR1 and NPFFR2) were expressed in cat ovaries by sequencing PCR products from ovarian RNA. We then isolated and cultured preantral ovarian follicles in the presence of 10 or 1 µM fRFRP3 + FSH (1 µg/mL). We recorded the percentage of morphologically viable follicles (basal lamina integrity) over 8 days and calculated percentage survival of follicles on Day 8 (using fluorescent markers for cell survival and death). Last, we quantified progesterone accumulation in media. 10 µM fRFRP3 had no observable effect on viability, survival, or steroid production compared to follicles exposed to only FSH. However, 1 µM fRFRP3 decreased the percentage of morphologically viable follicles and the percentage of surviving follicles on Day 8. At the same time, 1 µM fRFRP3 increased the accumulation of progesterone in media. Our study shows, for the first time, direct action of RFRP3 on the follicle as a functional unit, and it is the first in a carnivore species. More broadly, our results support a conserved, inhibitory action of RFRP3 on ovarian follicle development and underscore the importance of comparative functional studies.

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