Expression of muscarinic receptor types in the primate ovary and evidence for nonneuronal acetylcholine synthesis

S. Fritz, I. Wessler, R. Breitling, W. Rossmanith, Sergio Ojeda, Gregory Dissen, A. Amsterdam, A. Mayerhofer

    Research output: Contribution to journalArticle

    61 Citations (Scopus)

    Abstract

    The presence of muscarinic receptors (MR) in the ovary of different species has been recognized, but the identity of these receptors as well as ovarian sources of their natural ligand, acetylcholine (ACh), have not been determined. Because luteinized human granulosa cells (GC) in culture express functional MR, we have determined whether the group of the related MR subtypes, M1R, M3R, and M5R, are present in vivo in human and rhesus monkey ovaries. To this end, ribonucleic acids (RNAs) of different human and monkey ovaries as well as RNAs from human GC and monkey oocytes were reverse transcribed and subjected to PCR amplification, followed by sequencing of the amplified complementary DNAs. Results obtained showed that M1R, M3R, and M5R messenger RNAs are present in adult human and monkey ovaries; oocytes express exclusively the M3R subtype, whereas GC express M1R and M5R. To determine the ovarian source(s) of the natural ligand of these ACh receptors, we attempted to localize the enzyme responsible for its synthesis with the help of a monoclonal antibody recognizing choline acetyltransferase for immunohistochemistry. In neither human nor monkey sections did we detect immunoreactive choline acetyltransferase-positive fibers or nerve cells, but, surprisingly, GC of antral follicles showed prominent staining. To determine whether GC can produce ACh, human cultured GC derived from preovulatory follicles were analyzed using a high pressure liquid chromatography technique. The results showed that these cells contained ACh in concentrations ranging from 4.2-11.5 pmol/106 cells. Samples of a rat granulosa cell line likewise contained ACh. Thus, the ovary contains multiple MR, and GC of antral follicles are able to synthesize ACh, the ligand of MR. We propose that ACh may serve as an as yet unrecognized factor involved in the complex regulation of ovarian function in the primate, e.g. regulation of cell proliferation or progesterone production.

    Original languageEnglish (US)
    Pages (from-to)349-354
    Number of pages6
    JournalJournal of Clinical Endocrinology and Metabolism
    Volume86
    Issue number1
    DOIs
    StatePublished - 2001

    Fingerprint

    Granulosa Cells
    Muscarinic Receptors
    Primates
    Acetylcholine
    Ovary
    Haplorhini
    Choline O-Acetyltransferase
    RNA
    Ligands
    High pressure liquid chromatography
    Oocytes
    Cell proliferation
    Cholinergic Receptors
    Neurons
    Progesterone
    Amplification
    Rats
    Complementary DNA
    Macaca mulatta
    Monoclonal Antibodies

    ASJC Scopus subject areas

    • Biochemistry
    • Endocrinology, Diabetes and Metabolism

    Cite this

    Expression of muscarinic receptor types in the primate ovary and evidence for nonneuronal acetylcholine synthesis. / Fritz, S.; Wessler, I.; Breitling, R.; Rossmanith, W.; Ojeda, Sergio; Dissen, Gregory; Amsterdam, A.; Mayerhofer, A.

    In: Journal of Clinical Endocrinology and Metabolism, Vol. 86, No. 1, 2001, p. 349-354.

    Research output: Contribution to journalArticle

    Fritz, S. ; Wessler, I. ; Breitling, R. ; Rossmanith, W. ; Ojeda, Sergio ; Dissen, Gregory ; Amsterdam, A. ; Mayerhofer, A. / Expression of muscarinic receptor types in the primate ovary and evidence for nonneuronal acetylcholine synthesis. In: Journal of Clinical Endocrinology and Metabolism. 2001 ; Vol. 86, No. 1. pp. 349-354.
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    abstract = "The presence of muscarinic receptors (MR) in the ovary of different species has been recognized, but the identity of these receptors as well as ovarian sources of their natural ligand, acetylcholine (ACh), have not been determined. Because luteinized human granulosa cells (GC) in culture express functional MR, we have determined whether the group of the related MR subtypes, M1R, M3R, and M5R, are present in vivo in human and rhesus monkey ovaries. To this end, ribonucleic acids (RNAs) of different human and monkey ovaries as well as RNAs from human GC and monkey oocytes were reverse transcribed and subjected to PCR amplification, followed by sequencing of the amplified complementary DNAs. Results obtained showed that M1R, M3R, and M5R messenger RNAs are present in adult human and monkey ovaries; oocytes express exclusively the M3R subtype, whereas GC express M1R and M5R. To determine the ovarian source(s) of the natural ligand of these ACh receptors, we attempted to localize the enzyme responsible for its synthesis with the help of a monoclonal antibody recognizing choline acetyltransferase for immunohistochemistry. In neither human nor monkey sections did we detect immunoreactive choline acetyltransferase-positive fibers or nerve cells, but, surprisingly, GC of antral follicles showed prominent staining. To determine whether GC can produce ACh, human cultured GC derived from preovulatory follicles were analyzed using a high pressure liquid chromatography technique. The results showed that these cells contained ACh in concentrations ranging from 4.2-11.5 pmol/106 cells. Samples of a rat granulosa cell line likewise contained ACh. Thus, the ovary contains multiple MR, and GC of antral follicles are able to synthesize ACh, the ligand of MR. We propose that ACh may serve as an as yet unrecognized factor involved in the complex regulation of ovarian function in the primate, e.g. regulation of cell proliferation or progesterone production.",
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    AU - Fritz, S.

    AU - Wessler, I.

    AU - Breitling, R.

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