Progestin Signaling through an Olfactory G Protein and Membrane Progestin Receptor-α in Atlantic Croaker Sperm: Potential Role in Induction of Sperm Hypermotility

Tubbs, Christopher W.; Thomas, Peter

doi:10.1210/en.2008-0512

Cited by 71 publications

(76 citation statements)

References 57 publications

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“…During the reproductive cycle in mammals, fish and frogs mPRs, like mPRα, mediate the induction of oocyte maturation and sperm motility, which could involve the activation of a G-protein, upon binding of progestins (34,37,41,45). An immunological response linked to reproduction is the up-regulation of the leukemia inhibitory factor by progesterone, which is essential for the implantation and development of the embryo and inhibits several macrophage functions (25).…”

Section: Discussionmentioning

confidence: 99%

Purification of the goldfish membrane progestin receptor α (mPRα) expressedin yeast Pichia pastoris

Oshima

Nakayama

Roy³

et al. 2014

Biomed. Res.

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Membrane progestin receptors (mPRs) are key mediators of rapid, nongenomic actions of progestins on plasma membranes. We established a procedure for the expression and purification of recombinant goldfish mPRα using the methylotropic yeast Pichia pastoris. In P. pastoris, the recombinant protein, which carried C-terminal histidine and c-Myc tags, was expressed in an active form as the receptor for maturation-inducing steroids of fish. Expressed proteins were bound reversibly with a high affinity (K d = 9.4 nM) at a single binding site that could be saturated. After solubilization of mPRα with n-dodecyl-β-D-maltoside (DDM) from yeast membranes, the recombinant protein was purified using three different columns: first it was affinity-purified over nickelnitrilotriacetic acid (Ni-NTA), then bound to a cellulose resin with free amino groups and finally to a column with affinity for the c-Myc epitope. The identity of the purified protein was verified by MALDI-TOF/MS analysis and its capacity to bind progestin remained. Expression and purification of mPRα protein in its functional form will enable the screening of ligands and the determination of its three dimensional structure.Progestins are important steroids regulating final maturation at reproductive tissues in male and female vertebrates. They can be divided into natural and synthetic types. Natural type is found in humans and certain animals (32). Natural progestin in human is progesterone. In the medical fields, a variety of synthetic progestins are used. In teleost fishes, two distinct progestins have been identified as naturally occurring maturation inducing steroids (MIS), which control oocyte maturation. 17α,20β-dihydroxy-4-pregnen-3-one (17,20β-DHP) was identified in amago salmon (Oncorhynchus rhodurus) and 17α,20β,21-trihydroxy-4-pregnen-3-one (20β-S) was identified in Atlantic croaker (Micropogonias undulatus) and spotted seatrout (Cynoscion nebulous) (22). The structural differences between human progestin, progesterone, and fish progestins are that fish progestins have multiple hydroxyl groups on the side chain at positions 17, 20 and 21. In fish, MIS is produced by the follicular envelope in response to luteinizing hormone (LH) from the pituitary (15). In salmon, LH stimulates a number of signaling cascades leading to the production of 17α-hydroxyprogesterone (17α-HP) from progesterone and 17α-hydroxypregnenolone in the theca cells. Then, 17α-HP is converted to 17,20β-DHP by 20β-hydroxysteroid dehydrogenase (20β-HSD) in the granulosa cells (18,22). The membrane progestin receptor (mPR) mediates the nongenomic actions of progestins on the plasma membrane. It was identified in teleost, a ray-finned fish, at first and subsequently identified in other vertebrates including human (44,45). Research into the

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Section: Discussionmentioning

confidence: 99%

Purification of the goldfish membrane progestin receptor α (mPRα) expressedin yeast Pichia pastoris

Oshima

Nakayama

Roy³

et al. 2014

Biomed. Res.

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“…Previous studies have shown that fish sperm with low motility expressed decreased mPRa protein in their sperm membranes (Tubbs and Thomas 2009). Immunocytochemistry of whole croaker spermatozoa with anti-mPRa antibody suggested the potential role of the receptor in mediating the maturation-inducing steroid stimulation of sperm motility .…”

Section: Mpra Expression and Mouse Spermatogenesismentioning

confidence: 98%

Broad tissue expression of membrane progesterone receptor Alpha in normal mice

2010

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The broad tissue distribution of membrane progesterone receptor alpha (mPRalpha) in vertebrates suggests multiple physiological functions of the receptor. Current knowledge regarding the receptor distribution, however, is largely obtained via non-histological assays. In this study, the tissue distribution of mPRalpha in mice of both sexes was described using both histological and non-histological methods. Immunohistochemical analysis revealed that abundant expression of mPRalpha was consistently detected in the cytoplasm and membrane of smooth muscles in vasculatures, gastro-intestines, and uterus. It was also observed in myoepithelial cells of mammary gland and intra-ovarian myofibroblasts. These findings suggest that mPRalpha may function as a mediator of P4 in regulating function of smooth muscles or smooth muscle-like cells in numerous physiological processes such as vasodilation, transportation of contents within luminary organs, relaxation of the uterine myometrium during pregnancy, release of oocytes, and milk secretion. In addition, strong mPRalpha expression was identified in the parietal cells of gastric glands, indicating the potential roles of P4/mPRalpha signaling in the modulation of gastric acid secretion. Surprisingly, in the testis of male mice mPRalpha was mainly seen in the nuclei, rather than cytoplasm and/or membrane, of the primary and secondary spermatocytes, suggesting a direct role of the receptor in gene regulation. Our results indicate that mPRalpha may function as a key modulator of P4 in the modulation of multiple physiological functions in normal mice.

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“…The mPR␣ subtype is ubiquitously expressed in vertebrate tissues and is the intermediary in a variety of nongenomic progesterone actions in fishes, including induction of oocyte maturation and sperm motility (68,74) and inhibition of granulosa cell apoptosis (13). The mPR␣ has also been implicated in progesterone modulation of gonadotropin-releasing hormone (GnRH) neuronal, T lymphocyte, breast cancer cell, and myometrial cell functions in mammalian cells (10,11,23,36,59).…”

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confidence: 99%

Progesterone increases nitric oxide synthesis in human vascular endothelial cells through activation of membrane progesterone receptor-α

Pang

Dong

Thomas

2015

American Journal of Physiology-Endocrinology and Metabolism

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Pang Y, Dong J, Thomas P. Progesterone increases nitric oxide synthesis in human vascular endothelial cells through activation of membrane progesterone receptor-␣. Am J Physiol Endocrinol Metab 308: E899 -E911, 2015. First published March 24, 2015; doi:10.1152/ajpendo.00527.2014.-Progesterone exerts beneficial effects on the human cardiovascular system by inducing rapid increases in nitric oxide (NO) production in vascular endothelial cells, but the receptors mediating these nongenomic progesterone actions remain unclear. Using human umbilical vein endothelial cells (HUVECs) as a model, we show that progesterone binds to plasma membranes of HUVECs with the characteristics of membrane progesterone receptors (mPRs). The selective mPR agonist Org OD 02-0 had high binding affinity for the progesterone receptor on HUVEC membranes, whereas nuclear PR (nPR) agonists R5020 and medroxyprogesterone acetate displayed low binding affinities. Immunocytochemical and Western blot analyses confirmed that mPRs are expressed in HUVECs and are localized on their plasma membranes. NO levels increased rapidly after treatment with 20 nM progesterone, Org OD 02-0, and a progesterone-BSA conjugate but not with R5020, suggesting that this progesterone action is at the cell surface and initiated through mPRs. Progesterone and Org OD 02-0 (20 nM) also significantly increased endothelial nitric oxide synthase (eNOS) activity and eNOS phosphorylation. Knockdown of mPR␣ expression by treatment with small-interfering RNA (siRNA) blocked the stimulatory effects of 20 nM progesterone on NO production and eNOS phosphorylation, whereas knockdown of nPR was ineffective. Treatment with PI3K/ Akt and MAP kinase inhibitors blocked the stimulatory effects of progesterone, Org OD 02-0, and progesterone-BSA on NO production and eNOS phosphorylation and also prevented progesterone-and Org OD 02-0-induced increases in Akt and ERK phosphorylation. The results suggest that progesterone stimulation of NO production in HUVECs is mediated by mPR␣ and involves signaling through PI3K/Akt and MAP kinase pathways. membrane progesterone receptors; human vascular endothelial cells; rapid progesterone action; nitric oxide; nitric oxide synthase FEMALE REPRODUCTIVE HORMONES are widely considered to have beneficial effects on cardiovascular functions in women. The lower incidence of cardiovascular disease in middle-aged premenopausal women than in men and postmenopausal women has been attributed to the presence of female sex steroids in the circulation (22,27,41,47). Blood pressure is typically lower in women than in men and often drops during pregnancy when circulating levels of estrogens and progesterone are elevated (30,70,71). Furthermore, synthesis of a major regulator of vasodilation, nitric oxide (NO), is greater in women than in men (14). These sex differences in the occurrence of cardiovascular disease are due partly to the well-known beneficial effects of estrogens, which include upregulation of NO production in human endothelial cells (7,18,28,33). Numerous...

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Progestin Signaling through an Olfactory G Protein and Membrane Progestin Receptor-α in Atlantic Croaker Sperm: Potential Role in Induction of Sperm Hypermotility

Cited by 71 publications

References 57 publications

Purification of the goldfish membrane progestin receptor α (mPRα) expressedin yeast Pichia pastoris

Purification of the goldfish membrane progestin receptor α (mPRα) expressedin yeast Pichia pastoris

Broad tissue expression of membrane progesterone receptor Alpha in normal mice

Progesterone increases nitric oxide synthesis in human vascular endothelial cells through activation of membrane progesterone receptor-α

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