FSH regulates acetycholine production by ovarian granulosa cells

Mayerhofer, Artur; Kunz, Lars; Krieger, Annette; Proskocil, Becky J.; Spindel, Eliot R.; Amsterdam, Abraham; Dissen, Gregory A.; Ojeda, Sergio R.; Wessler, Ignaz

doi:10.1186/1477-7827-4-37

Cited by 35 publications

(32 citation statements)

References 30 publications

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“…The specificity of the CHAT antibody used in the present study was confirmed previously by its reactivity with cholinergic neural cells and fibers in the bovine tissues [33]. These findings are in agreement with those of previous studies that demonstrated CHAT expression and/or ACH production in endocrine tissues, such as ovarian granulosa cells of healthy follicles in humans, monkeys, and rats [9][10][11]; spermatogenic cells and mature spermatozoa in the rat testis [14]; and syncytiotrophoblast cells in both human and rat placenta [12,13].…”

Section: Discussionsupporting

confidence: 94%

“…De novo synthesis and ACH release have been demonstrated in the granulosa cells of healthy follicles in primates, humans, and rats [8][9][10][11]. A nonneural source of ACH has been demonstrated in various cell types in the body, such as syncytiotrophoblast and amnion epithelial cells in the placenta [12,13]; spermatogenic cells in the testis [14]; gastrointestinal, respiratory, and urogenital epithelial cells [15][16][17]; and immune cells, including lymphocytes [18], mucocutaneous epithelial cells and keratinocytes [19], and vascular endothelial cells [20].…”

Section: Introductionmentioning

confidence: 99%

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Survival Role of Locally Produced Acetylcholine in the Bovine Corpus Luteum1

Al-zi’abi

Bowolaksono

Okuda

2009

Biology of Reproduction

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The present study was conducted to explore the source of acetylcholine (ACH) in the corpus luteum (CL) and to test our hypothesis of an antiapoptotic role of ACH in the bovine CL and, further, to investigate whether nerve growth factor (NGF), insulin-like growth factor 1 (IGF1), and transforming growth factor beta1 (TGFB1) influence the expression of choline acetyltransferase (CHAT), the biosynthetic enzyme of ACH, in cultured bovine luteal cells. Protein expression and immunolocalization of CHAT were carried out at different stages throughout the luteal phase and in cultured luteal and endothelial cells. ACH was measured in luteal tissue at the different luteal stages and in luteal cells cultured for 8 and 24 h. Cell viability and TUNEL assays were performed on cultured midluteal cells treated with or without tumor necrosis factor alpha (TNF)/interferon gamma (IFNG) in the presence of ACH and its muscarinic (atropine) and nicotinic (mecamylamine) receptor antagonists. The CL was devoid of cholinergic nerve fibers. CHAT immunostaining was evident in luteal, endothelial, and stromal cells in luteal tissue sections and in cultured luteal and endothelial cells. CHAT protein was expressed throughout the cycle without any significant changes. ACH concentration in luteal tissue was not changed during the luteal stages but increased over time and with increased cell numbers in luteal cell cultures. ACH increased cell viability and prevented cell death induced by TNF/IFNG. Atropine significantly attenuated ACH action, whereas mecamylamine had no effect. TNF/IFNG treatment downregulated CHAT expression, whereas NGF, IGF1, and TGFB1 upregulated CHAT expression, in cultured luteal cells. The overall findings strongly suggest a nonneural source and antiapoptotic role of ACH in the bovine CL. Locally produced ACH appears to be regulated by NGF, IGF1, and TGFB1.

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Section: Discussionsupporting

confidence: 94%

Section: Introductionmentioning

confidence: 99%

Survival Role of Locally Produced Acetylcholine in the Bovine Corpus Luteum1

Al-zi’abi

Bowolaksono

Okuda

2009

Biology of Reproduction

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“…Monkey ovarian samples from Rhesus macaques ( Macaca Mulatta , age 5-6 years) were the same as used in previous studies [29]. Collection of tissues was approved by the Oregon National Primate Research Center Institutional Animal Care and Use Committee.…”

Section: Methodsmentioning

confidence: 99%

Pro-nerve growth factor in the ovary and human granulosa cells

Meinel

Blohberger

Berg

et al. 2015

Hormone Molecular Biology and Clinical Investigation

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Background Pro-nerve growth factor must be cleaved to generate mature NGF, which was suggested to be a factor involved in ovarian physiology and pathology. Extracellular proNGF can induce cell death in many tissues. Whether extracellular proNGF exists in the ovary and may play a role in the death of follicular cells or atresia was unknown. Material and Methods Immunohistochemistry of human and Rhesus monkey ovarian sections was performed. IVF-derived follicular fluid and human granulosa cells were studied by RT-PCR, qPCR, Western blotting, ATP- and caspase-assays. Results and Conclusions Immunohistochemistry of ovarian sections identified proNGF in granulosa cells and Western blotting of human isolated granulosa cells confirmed the presence of proNGF. Ovarian granulosa cells thus produce proNGF. Recombinant human proNGF even at high concentrations did not affect the levels of ATP or the activity of caspase 3/7, indicating that in granulosa cells proNGF does not induce death. In contrast, mature NGF, which was detected previously in follicular fluid, may be a trophic molecule for granulosa cells with unexpected functions. We found that in contrast to proNGF, NGF increased the levels of the transcription factor early growth response 1 and of the enzyme choline acetyl-transferase. A mechanism for the generation of mature NGF from proNGF in the follicular fluid may be extracellular enzymatic cleavage. The enzyme MMP7 is known to cleave proNGF and was identified in follicular fluid and as a product of granulosa cells. Thus the generation of NGF in the ovarian follicle may depend on MMP7.

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“…ACh was either determined using the HPLC method mentioned above (Fritz et al 1999(Fritz et al , 2001 or a fluorimetricbased method (Amplex Red Acetylcholine Assay kit, Molecular Probes/Invitrogen GmbH, Karlsruhe, Germany). Details have recently been provided (Mayerhofer et al 2006). All measurements were performed in triplicates.…”

Section: Ach Measurementmentioning

confidence: 99%

Non-neuronal acetylcholine and choline acetyltransferase in oviductal epithelial cells of cyclic and pregnant pigs

et al. 2006

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Certain female reproductive tissues are known to express the non-neuronal cholinergic system. Using different experimental approaches, we tested the hypothesis that acetylcholine (ACh) in the porcine oviduct may also be derived from non-neuronal structures. Immunohistochemistry was performed to detect acetylcholine synthesizing enzyme choline acetyltransferase (ChAT) in different segments of the oviduct of cyclic and pregnant sows. Immunohistochemical experiments revealed strong immunoexpression of ChAT in the entire oviductal epithelium at metoestrus. Thereby, a particular pronounced staining was found in the supranuclear region of almost all epithelial cells. Immunostaining of ChAT decreased markedly during dioestrus and prooestrus stages, respectively. At prooestrus, ChAT immunoreactivity was confined to ciliated cells. Furthermore, we found elevated level of staining intensity of ChAT in the pregnant oviduct at day 13. Using the same ChAT antibody for Western blot analyses, we detected immunoreactive bands of MW 69,000 and 46,000 mainly in ampulla, while MW 58,000 and 30,000 forms were present mainly in infundibulum and isthmus. Furthermore ACh was detected by HPLC and fluorimetric methods in oviductal epithelium. In conclusion, we show expression of ChAT in oviductal epithelial cells at different stages of the oestrus cycle and pregnancy, indicating that these cells can synthesize ACh in a cycle-dependent manner. These results suggest as yet unexplored roles of epithelial ACh in the oviduct.

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FSH regulates acetycholine production by ovarian granulosa cells

Cited by 35 publications

References 30 publications

Survival Role of Locally Produced Acetylcholine in the Bovine Corpus Luteum1

Survival Role of Locally Produced Acetylcholine in the Bovine Corpus Luteum1

Pro-nerve growth factor in the ovary and human granulosa cells

Non-neuronal acetylcholine and choline acetyltransferase in oviductal epithelial cells of cyclic and pregnant pigs

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