Expression of parathyroid hormone-related peptide (PTH-rp) and its receptor in the porcine ovary: Regulation by transforming growth factor-β and possible paracrine effects of granulosa cell PTH-rp secretion on theca cells

J. C. Garmey, J. A. Schnorr, M. E. Bruns, D. E. Bruns, R. M. Seaner, J. E. Ferguson, F. C L Jayes, C. Aguirre, Johannes D Veldhuis

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Parathyroid hormone-related peptide (PTH-rp) and the PTH-rp receptor are expressed in certain cancers as well as in many normal tissues. To evaluate the expression of this Ca 2+-regulating hormone and its receptor in porcine ovary, we isolated partial cDNAs encoding homologous PTH-rp and PTH-rp receptor using reverse transcription-polymerase chain reaction (RT-PCR). The cDNA encoding PTH-rp (419 base pairs [bp]) was 92% and 87% homologous to human and rat sequences, respectively, while the PTH-rp receptor clone (167 bp) was 94% and 91% identical to the human and rat genes. Qualitative estimates of PTH-rp mRNA by RT-PCR indicated that the PTH-rp gene is expressed at high levels in the corpus luteum but is undetectable in granulosa and theca cells isolated from small (1-5 mm) and medium-sized (5-8 mm) antral follicles. In contrast, PTH-rp receptor transcripts were most abundant in corpora lutea and theca cells, and least abundant (albeit detectable) in granulosa cells. Regulation of PTH-rp protein production was assessed in serum-free monolayer cultures of porcine granulosa cells. Transforming growth factor (TGF)-β1 (100 ng/ml) increased PTH-rp concentrations (assayed by two-site immunoradiometric assay of culture media) as well as corresponding PTH-rp mRNA accumulation (assessed by RT-PCR) in a time-dependent manner, with maximal responses of 3- to 5-fold at 96 h. TGF- β1 dose-response studies revealed an ED 50 of 0.24-0.38 ng/ml with a maximal effect at 30 ng/ml. Other growth factors and hormones, including insulin, insulin-like growth factor (type I), epidermal growth factor, FSH, estradiol, and interleukin-1, failed to alter PTH-rp secretion. Biological effects of PTH-rp were evident in purified porcine theca cells. Using the Ca 2+-sensitive fluorescent indicator dye, fura-2, and digital imaging videomicroscopy, we found that PTHrp (1 μM) stimulated intracellular free calcium ion concentrations ([Ca 2+](i)) in single porcine theca cells. The [Ca 2+](i) elevation was characterized by a slow and prolonged rise. After PTH-rp stimulation, theca cells maintained responsiveness to hormone stimulation by LH, which elicited a typical theca cell [Ca 2+](i) response. Our results allow a hypothesis of a paracrine intrafollicular signaling system involving interaction between theca cell-derived TGF-β and granulosa cell-derived PTH-rp, with feedback by PTH-rp on theca cells. Alternatively, expression of mRNAs encoding PTH-rp and its receptor in corpora lutea suggests that this peptide may play a role in luteal cell function. The precise role of this intraovarian PTH-rp system will require further study.

Original languageEnglish (US)
Pages (from-to)334-339
Number of pages6
JournalBiology of Reproduction
Volume62
Issue number2
StatePublished - 2000
Externally publishedYes

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Parathyroid Hormone Receptor Type 1
Theca Cells
Parathyroid Hormone-Related Protein
Granulosa Cells
Transforming Growth Factors
Ovary
Swine
Corpus Luteum
Reverse Transcription
Peptide Hormones
Base Pairing
Polymerase Chain Reaction
Messenger RNA
Complementary DNA
Paracrine Communication
Hormones
Immunoradiometric Assay
Luteal Cells
Video Microscopy
Fura-2

ASJC Scopus subject areas

  • Cell Biology
  • Developmental Biology
  • Embryology

Cite this

Expression of parathyroid hormone-related peptide (PTH-rp) and its receptor in the porcine ovary : Regulation by transforming growth factor-β and possible paracrine effects of granulosa cell PTH-rp secretion on theca cells. / Garmey, J. C.; Schnorr, J. A.; Bruns, M. E.; Bruns, D. E.; Seaner, R. M.; Ferguson, J. E.; Jayes, F. C L; Aguirre, C.; Veldhuis, Johannes D.

In: Biology of Reproduction, Vol. 62, No. 2, 2000, p. 334-339.

Research output: Contribution to journalArticle

Garmey, J. C. ; Schnorr, J. A. ; Bruns, M. E. ; Bruns, D. E. ; Seaner, R. M. ; Ferguson, J. E. ; Jayes, F. C L ; Aguirre, C. ; Veldhuis, Johannes D. / Expression of parathyroid hormone-related peptide (PTH-rp) and its receptor in the porcine ovary : Regulation by transforming growth factor-β and possible paracrine effects of granulosa cell PTH-rp secretion on theca cells. In: Biology of Reproduction. 2000 ; Vol. 62, No. 2. pp. 334-339.
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abstract = "Parathyroid hormone-related peptide (PTH-rp) and the PTH-rp receptor are expressed in certain cancers as well as in many normal tissues. To evaluate the expression of this Ca 2+-regulating hormone and its receptor in porcine ovary, we isolated partial cDNAs encoding homologous PTH-rp and PTH-rp receptor using reverse transcription-polymerase chain reaction (RT-PCR). The cDNA encoding PTH-rp (419 base pairs [bp]) was 92{\%} and 87{\%} homologous to human and rat sequences, respectively, while the PTH-rp receptor clone (167 bp) was 94{\%} and 91{\%} identical to the human and rat genes. Qualitative estimates of PTH-rp mRNA by RT-PCR indicated that the PTH-rp gene is expressed at high levels in the corpus luteum but is undetectable in granulosa and theca cells isolated from small (1-5 mm) and medium-sized (5-8 mm) antral follicles. In contrast, PTH-rp receptor transcripts were most abundant in corpora lutea and theca cells, and least abundant (albeit detectable) in granulosa cells. Regulation of PTH-rp protein production was assessed in serum-free monolayer cultures of porcine granulosa cells. Transforming growth factor (TGF)-β1 (100 ng/ml) increased PTH-rp concentrations (assayed by two-site immunoradiometric assay of culture media) as well as corresponding PTH-rp mRNA accumulation (assessed by RT-PCR) in a time-dependent manner, with maximal responses of 3- to 5-fold at 96 h. TGF- β1 dose-response studies revealed an ED 50 of 0.24-0.38 ng/ml with a maximal effect at 30 ng/ml. Other growth factors and hormones, including insulin, insulin-like growth factor (type I), epidermal growth factor, FSH, estradiol, and interleukin-1, failed to alter PTH-rp secretion. Biological effects of PTH-rp were evident in purified porcine theca cells. Using the Ca 2+-sensitive fluorescent indicator dye, fura-2, and digital imaging videomicroscopy, we found that PTHrp (1 μM) stimulated intracellular free calcium ion concentrations ([Ca 2+](i)) in single porcine theca cells. The [Ca 2+](i) elevation was characterized by a slow and prolonged rise. After PTH-rp stimulation, theca cells maintained responsiveness to hormone stimulation by LH, which elicited a typical theca cell [Ca 2+](i) response. Our results allow a hypothesis of a paracrine intrafollicular signaling system involving interaction between theca cell-derived TGF-β and granulosa cell-derived PTH-rp, with feedback by PTH-rp on theca cells. Alternatively, expression of mRNAs encoding PTH-rp and its receptor in corpora lutea suggests that this peptide may play a role in luteal cell function. The precise role of this intraovarian PTH-rp system will require further study.",
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T1 - Expression of parathyroid hormone-related peptide (PTH-rp) and its receptor in the porcine ovary

T2 - Regulation by transforming growth factor-β and possible paracrine effects of granulosa cell PTH-rp secretion on theca cells

AU - Garmey, J. C.

AU - Schnorr, J. A.

AU - Bruns, M. E.

AU - Bruns, D. E.

AU - Seaner, R. M.

AU - Ferguson, J. E.

AU - Jayes, F. C L

AU - Aguirre, C.

AU - Veldhuis, Johannes D

PY - 2000

Y1 - 2000

N2 - Parathyroid hormone-related peptide (PTH-rp) and the PTH-rp receptor are expressed in certain cancers as well as in many normal tissues. To evaluate the expression of this Ca 2+-regulating hormone and its receptor in porcine ovary, we isolated partial cDNAs encoding homologous PTH-rp and PTH-rp receptor using reverse transcription-polymerase chain reaction (RT-PCR). The cDNA encoding PTH-rp (419 base pairs [bp]) was 92% and 87% homologous to human and rat sequences, respectively, while the PTH-rp receptor clone (167 bp) was 94% and 91% identical to the human and rat genes. Qualitative estimates of PTH-rp mRNA by RT-PCR indicated that the PTH-rp gene is expressed at high levels in the corpus luteum but is undetectable in granulosa and theca cells isolated from small (1-5 mm) and medium-sized (5-8 mm) antral follicles. In contrast, PTH-rp receptor transcripts were most abundant in corpora lutea and theca cells, and least abundant (albeit detectable) in granulosa cells. Regulation of PTH-rp protein production was assessed in serum-free monolayer cultures of porcine granulosa cells. Transforming growth factor (TGF)-β1 (100 ng/ml) increased PTH-rp concentrations (assayed by two-site immunoradiometric assay of culture media) as well as corresponding PTH-rp mRNA accumulation (assessed by RT-PCR) in a time-dependent manner, with maximal responses of 3- to 5-fold at 96 h. TGF- β1 dose-response studies revealed an ED 50 of 0.24-0.38 ng/ml with a maximal effect at 30 ng/ml. Other growth factors and hormones, including insulin, insulin-like growth factor (type I), epidermal growth factor, FSH, estradiol, and interleukin-1, failed to alter PTH-rp secretion. Biological effects of PTH-rp were evident in purified porcine theca cells. Using the Ca 2+-sensitive fluorescent indicator dye, fura-2, and digital imaging videomicroscopy, we found that PTHrp (1 μM) stimulated intracellular free calcium ion concentrations ([Ca 2+](i)) in single porcine theca cells. The [Ca 2+](i) elevation was characterized by a slow and prolonged rise. After PTH-rp stimulation, theca cells maintained responsiveness to hormone stimulation by LH, which elicited a typical theca cell [Ca 2+](i) response. Our results allow a hypothesis of a paracrine intrafollicular signaling system involving interaction between theca cell-derived TGF-β and granulosa cell-derived PTH-rp, with feedback by PTH-rp on theca cells. Alternatively, expression of mRNAs encoding PTH-rp and its receptor in corpora lutea suggests that this peptide may play a role in luteal cell function. The precise role of this intraovarian PTH-rp system will require further study.

AB - Parathyroid hormone-related peptide (PTH-rp) and the PTH-rp receptor are expressed in certain cancers as well as in many normal tissues. To evaluate the expression of this Ca 2+-regulating hormone and its receptor in porcine ovary, we isolated partial cDNAs encoding homologous PTH-rp and PTH-rp receptor using reverse transcription-polymerase chain reaction (RT-PCR). The cDNA encoding PTH-rp (419 base pairs [bp]) was 92% and 87% homologous to human and rat sequences, respectively, while the PTH-rp receptor clone (167 bp) was 94% and 91% identical to the human and rat genes. Qualitative estimates of PTH-rp mRNA by RT-PCR indicated that the PTH-rp gene is expressed at high levels in the corpus luteum but is undetectable in granulosa and theca cells isolated from small (1-5 mm) and medium-sized (5-8 mm) antral follicles. In contrast, PTH-rp receptor transcripts were most abundant in corpora lutea and theca cells, and least abundant (albeit detectable) in granulosa cells. Regulation of PTH-rp protein production was assessed in serum-free monolayer cultures of porcine granulosa cells. Transforming growth factor (TGF)-β1 (100 ng/ml) increased PTH-rp concentrations (assayed by two-site immunoradiometric assay of culture media) as well as corresponding PTH-rp mRNA accumulation (assessed by RT-PCR) in a time-dependent manner, with maximal responses of 3- to 5-fold at 96 h. TGF- β1 dose-response studies revealed an ED 50 of 0.24-0.38 ng/ml with a maximal effect at 30 ng/ml. Other growth factors and hormones, including insulin, insulin-like growth factor (type I), epidermal growth factor, FSH, estradiol, and interleukin-1, failed to alter PTH-rp secretion. Biological effects of PTH-rp were evident in purified porcine theca cells. Using the Ca 2+-sensitive fluorescent indicator dye, fura-2, and digital imaging videomicroscopy, we found that PTHrp (1 μM) stimulated intracellular free calcium ion concentrations ([Ca 2+](i)) in single porcine theca cells. The [Ca 2+](i) elevation was characterized by a slow and prolonged rise. After PTH-rp stimulation, theca cells maintained responsiveness to hormone stimulation by LH, which elicited a typical theca cell [Ca 2+](i) response. Our results allow a hypothesis of a paracrine intrafollicular signaling system involving interaction between theca cell-derived TGF-β and granulosa cell-derived PTH-rp, with feedback by PTH-rp on theca cells. Alternatively, expression of mRNAs encoding PTH-rp and its receptor in corpora lutea suggests that this peptide may play a role in luteal cell function. The precise role of this intraovarian PTH-rp system will require further study.

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