PI3K signalling in GnRH actions on dispersed goldfish pituitary cells: Relationship with PKC-mediated LH and GH release and regulation of long-term effects on secretion and total cellular hormone availability

Pemberton, Joshua; Orr, Michael E.; Stafford, James L.; Chang, John P.

doi:10.1016/j.ygcen.2014.03.011

Cited by 11 publications

(6 citation statements)

References 54 publications

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“…Due to the documented role of PI3Ks in fish pituitary hormone secretion ( 14 , 54 , 55 ) and their inhibition by Wm, we initially hypothesized that Wm blockade of PI3KCs in lactotrophs caused inhibition of PRL release. We tested this hypothesis in two ways.…”

Section: Discussionmentioning

confidence: 99%

Calcium-Prolactin Secretion Coupling in Rat Pituitary Lactotrophs Is Controlled by PI4-Kinase Alpha

et al. 2021

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The role of calcium, but not of other intracellular signaling molecules, in the release of pituitary hormones by exocytosis is well established. Here, we analyzed the contribution of phosphatidylinositol kinases (PIKs) to calcium-driven prolactin (PRL) release in pituitary lactotrophs: PI4Ks - which control PI4P production, PIP5Ks - which synthesize PI(4, 5)P2 by phosphorylating the D-5 position of the inositol ring of PI4P, and PI3KCs – which phosphorylate PI(4, 5)P2 to generate PI(3, 4, 5)P3. We used common and PIK-specific inhibitors to evaluate the strength of calcium-secretion coupling in rat lactotrophs. Gene expression was analyzed by single-cell RNA sequencing and qRT-PCR analysis; intracellular and released hormones were assessed by radioimmunoassay and ELISA; and single-cell calcium signaling was recorded by Fura 2 imaging. Single-cell RNA sequencing revealed the expression of Pi4ka, Pi4kb, Pi4k2a, Pi4k2b, Pip5k1a, Pip5k1c, and Pik3ca, as well as Pikfyve and Pip4k2c, in lactotrophs. Wortmannin, a PI3K and PI4K inhibitor, but not LY294002, a PI3K inhibitor, blocked spontaneous action potential driven PRL release with a half-time of ~20 min when applied in 10 µM concentration, leading to accumulation of intracellular PRL content. Wortmannin also inhibited increase in PRL release by high potassium, the calcium channel agonist Bay K8644, and calcium mobilizing thyrotropin-releasing hormone without affecting accompanying calcium signaling. GSK-A1, a specific inhibitor of PI4KA, also inhibited calcium-driven PRL secretion without affecting calcium signaling and Prl expression. In contrast, PIK93, a specific inhibitor of PI4KB, and ISA2011B and UNC3230, specific inhibitors of PIP5K1A and PIP5K1C, respectively, did not affect PRL release. These experiments revealed a key role of PI4KA in calcium-secretion coupling in pituitary lactotrophs downstream of voltage-gated and PI(4, 5)P2-dependent calcium signaling.

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Section: Discussionmentioning

confidence: 99%

Calcium-Prolactin Secretion Coupling in Rat Pituitary Lactotrophs Is Controlled by PI4-Kinase Alpha

et al. 2021

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“…Sanger sequencing validated the back-spliced junction of rno_circ_0001004 (Figure 1B). protein secretion including GH and luteinizing hormone (LH) [13,14], as well as regulating cell proliferation [15]. Circular RNA (circRNA) is a special type of non-coding RNA (ncRNA) molecule that, unlike traditional linear RNA, forms covalently closed loop structures generated by pre-mRNA back splicing.…”

Section: Characterization Of Rno_circ_0001004 In Gh 3 Cellsmentioning

confidence: 99%

“…We have previously shown that the miR-709 is highly expressed in the pituitary and inhibits the GH synthesis and suppresses the viability of GH 3 cells [12] by targeting Protein Kinase C alpha (PKCA). Protein Kinase C (PKC) is a class of phospholipid-dependent kinases that participate in regulation of protein secretion including GH and luteinizing hormone (LH) [13,14], as well as regulating cell proliferation [15].…”

Section: Introductionmentioning

confidence: 99%

Rno_circ_0001004 Acts as a miR-709 Molecular Sponge to Regulate the Growth Hormone Synthesis and Cell Proliferation

Xiong

Zhang

Wang

et al. 2022

IJMS

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(1) Background: As a novel type of non-coding RNA with a stable closed-loop structure, circular RNA (circRNA) can interact with microRNA (miRNA) and influence the expression of miRNA target genes. However, circRNA involved in pituitary growth hormone (GH) regulation is poorly understood. Our previous study revealed protein kinase C alpha (PRKCA) as the target gene of miR-709. Currently, the expression and function of rno_circRNA_0001004 in the rat pituitary gland is not clarified; (2) Methods: In this study, both bioinformatics analysis and dual-luciferase report assays showed a target relationship between rno_circRNA_0001004 and miR-709. Furthermore, the rno_circRNA_0001004 overexpression vector and si-circ_0001004 were constructed and transfected into GH3 cells; (3) Results: We found that rno_circRNA_0001004 expression was positively correlated with the PRKCA gene and GH expression levels, while it was negatively correlated with miR-709. In addition, overexpression of rno-circ_0001004 also promoted proliferation and relieved the inhibition of miR-709 in GH3 cells; (4) Conclusions: Our findings show that rno_circ_0001004 acts as a novel sponge for miR-709 to regulate GH synthesis and cell proliferation, and are the first case of discovery of the regulatory role of circRNA_0001004 in pituitary GH.

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“…Dispersed goldfish pituitary cells were cultured overnight in plating media (4.5 Â 10 6 cells in centrifuge-safe tissue culture tubes (Techno Plastic Products (TPP) T-10; Midwest Scientific; Valley Park, MO, USA) as previously described in Pemberton et al (2014). On the next day, cells were centrifuged (200Âg, 10 min) and washed with imaging media, and incubated with 1 lM DAF-FM DA (4-amino-5-methylamino-2 0 ,7 0 -difluorofluorescein diacetate; Molecular Probes, Life Technologies) in imaging media for 45 min in the incubator to allowing for deacylation of DAF-FM DA by intracellular esterases and trapping of the now membrane impermeant DAF-FM within cells.…”

Section: No-induced Fluorescence In Goldfish Somatotropesmentioning

confidence: 99%

“…Furthermore phosphoinositide 3-kinase (PI3K) signalling also mediates GnRH2 and GnRH3 stimulation of GH secretion (Pemberton et al, 2011(Pemberton et al, , 2014. Interestingly, NO is also known to have negative and positive modulating influences on Ca 2+ release through IP 3 -sensitive Ca 2+ channels in vascular and bronchial smooth muscles, respectively (Perez-Zoghbi et al, 2010;Song et al, 2011).…”

Section: Implications On the Integration Of Intracellular Signal Tranmentioning

confidence: 99%

Relationship between nitric oxide- and calcium-dependent signal transduction pathways in growth hormone release from dispersed goldfish pituitary cells

Chang

Sawisky

Davis

et al. 2014

General and Comparative Endocrinology

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PI3K signalling in GnRH actions on dispersed goldfish pituitary cells: Relationship with PKC-mediated LH and GH release and regulation of long-term effects on secretion and total cellular hormone availability

Cited by 11 publications

References 54 publications

Calcium-Prolactin Secretion Coupling in Rat Pituitary Lactotrophs Is Controlled by PI4-Kinase Alpha

Calcium-Prolactin Secretion Coupling in Rat Pituitary Lactotrophs Is Controlled by PI4-Kinase Alpha

Rno_circ_0001004 Acts as a miR-709 Molecular Sponge to Regulate the Growth Hormone Synthesis and Cell Proliferation

Relationship between nitric oxide- and calcium-dependent signal transduction pathways in growth hormone release from dispersed goldfish pituitary cells

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