Dopamine receptor repertoire of human granulosa cells
<p>Abstract</p> <p>Background</p> <p>High levels of dopamine (DA) were described in human ovary and recently evidence for DA receptors in granulosa and luteal cells has been provided, as well. However, neither the full repertoire of ovarian receptors for DA, nor their s...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2007-10-01
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| Series: | Reproductive Biology and Endocrinology |
| Online Access: | http://www.rbej.com/content/5/1/40 |
| _version_ | 1811260493910769664 |
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| author | Kunz Lars Berg Ulrike Berg Dieter Lazarov Nikolai Rey-Ares Veronica Mayerhofer Artur |
| author_facet | Kunz Lars Berg Ulrike Berg Dieter Lazarov Nikolai Rey-Ares Veronica Mayerhofer Artur |
| author_sort | Kunz Lars |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>High levels of dopamine (DA) were described in human ovary and recently evidence for DA receptors in granulosa and luteal cells has been provided, as well. However, neither the full repertoire of ovarian receptors for DA, nor their specific role, is established. Human granulosa cells (GCs) derived from women undergoing in vitro fertilization (IVF) are an adequate model for endocrine cells of the follicle and the corpus luteum and were therefore employed in an attempt to decipher their DA receptor repertoire and functionality.</p> <p>Methods</p> <p>Cells were obtained from patients undergoing IVF and examined using cDNA-array, RT-PCR, Western blotting and immunocytochemistry. In addition, calcium measurements (with FLUO-4) were employed. Expression of two DA receptors was also examined by in-situ hybridization in rat ovary. Effects of DA on cell viability and cell volume were studied by using an ATP assay and an electronic cell counter system.</p> <p>Results</p> <p>We found members of the two DA receptor families (D<sub>1</sub>- and D<sub>2</sub> -like) associated with different signaling pathways in human GCs, namely D<sub>1</sub> (as expected) and D<sub>5</sub> (both are Gs coupled and linked to cAMP increase) and D<sub>2</sub>, D<sub>4</sub> (Gi/Gq coupled and linked to IP3/DAG). D<sub>3</sub> was not found. The presence of the trophic hormone hCG (10 IU/ml) in the culture medium for several days did not alter mRNA (semiquantitative RT-PCR) or protein levels (immunocytochemistry/Western blotting) of D<sub>1,2,4,5</sub> DA receptors. Expression of prototype receptors for the two families, D<sub>1</sub> and D<sub>2</sub>, was furthermore shown in rat granulosa and luteal cells by in situ hybridization. Among the DA receptors found in human GCs, D<sub>2</sub> expression was marked both at mRNA and protein levels and it was therefore further studied. Results of additional RT-PCR and Western blots showed two splice variants (D<sub>2</sub>L, D<sub>2</sub>S). Irrespective of these variants, D<sub>2</sub> proved to be functional, as DA raised intracellular calcium levels. This calcium mobilizing effect of DA was observed in the absence of extracellular calcium and was abolished by a D<sub>2</sub> blocker (L-741,626). DA treatment (48 h) of human GCs resulted in slightly, but significantly enlarged, viable cells.</p> <p>Conclusion</p> <p>A previous study showed D<sub>2</sub> in human GCs, which are linked to cAMP, and the present study reveals the full spectrum of DA receptors present in these endocrine cells, which also includes D<sub>2</sub>-like receptors, linked to calcium. Ovarian DA can act thus via D<sub>1,2,4,5</sub>, which are co-expressed by endocrine cells of the follicle and the corpus luteum and are linked to different signaling pathways. This suggests a complex role of DA in the regulation of ovarian processes.</p> |
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| format | Article |
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| institution | Directory Open Access Journal |
| issn | 1477-7827 |
| language | English |
| last_indexed | 2024-04-12T18:48:08Z |
| publishDate | 2007-10-01 |
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| series | Reproductive Biology and Endocrinology |
| spelling | doaj.art-82b383c0b68146fe8ddb998efcb62bd02022-12-22T03:20:33ZengBMCReproductive Biology and Endocrinology1477-78272007-10-01514010.1186/1477-7827-5-40Dopamine receptor repertoire of human granulosa cellsKunz LarsBerg UlrikeBerg DieterLazarov NikolaiRey-Ares VeronicaMayerhofer Artur<p>Abstract</p> <p>Background</p> <p>High levels of dopamine (DA) were described in human ovary and recently evidence for DA receptors in granulosa and luteal cells has been provided, as well. However, neither the full repertoire of ovarian receptors for DA, nor their specific role, is established. Human granulosa cells (GCs) derived from women undergoing in vitro fertilization (IVF) are an adequate model for endocrine cells of the follicle and the corpus luteum and were therefore employed in an attempt to decipher their DA receptor repertoire and functionality.</p> <p>Methods</p> <p>Cells were obtained from patients undergoing IVF and examined using cDNA-array, RT-PCR, Western blotting and immunocytochemistry. In addition, calcium measurements (with FLUO-4) were employed. Expression of two DA receptors was also examined by in-situ hybridization in rat ovary. Effects of DA on cell viability and cell volume were studied by using an ATP assay and an electronic cell counter system.</p> <p>Results</p> <p>We found members of the two DA receptor families (D<sub>1</sub>- and D<sub>2</sub> -like) associated with different signaling pathways in human GCs, namely D<sub>1</sub> (as expected) and D<sub>5</sub> (both are Gs coupled and linked to cAMP increase) and D<sub>2</sub>, D<sub>4</sub> (Gi/Gq coupled and linked to IP3/DAG). D<sub>3</sub> was not found. The presence of the trophic hormone hCG (10 IU/ml) in the culture medium for several days did not alter mRNA (semiquantitative RT-PCR) or protein levels (immunocytochemistry/Western blotting) of D<sub>1,2,4,5</sub> DA receptors. Expression of prototype receptors for the two families, D<sub>1</sub> and D<sub>2</sub>, was furthermore shown in rat granulosa and luteal cells by in situ hybridization. Among the DA receptors found in human GCs, D<sub>2</sub> expression was marked both at mRNA and protein levels and it was therefore further studied. Results of additional RT-PCR and Western blots showed two splice variants (D<sub>2</sub>L, D<sub>2</sub>S). Irrespective of these variants, D<sub>2</sub> proved to be functional, as DA raised intracellular calcium levels. This calcium mobilizing effect of DA was observed in the absence of extracellular calcium and was abolished by a D<sub>2</sub> blocker (L-741,626). DA treatment (48 h) of human GCs resulted in slightly, but significantly enlarged, viable cells.</p> <p>Conclusion</p> <p>A previous study showed D<sub>2</sub> in human GCs, which are linked to cAMP, and the present study reveals the full spectrum of DA receptors present in these endocrine cells, which also includes D<sub>2</sub>-like receptors, linked to calcium. Ovarian DA can act thus via D<sub>1,2,4,5</sub>, which are co-expressed by endocrine cells of the follicle and the corpus luteum and are linked to different signaling pathways. This suggests a complex role of DA in the regulation of ovarian processes.</p>http://www.rbej.com/content/5/1/40 |
| spellingShingle | Kunz Lars Berg Ulrike Berg Dieter Lazarov Nikolai Rey-Ares Veronica Mayerhofer Artur Dopamine receptor repertoire of human granulosa cells Reproductive Biology and Endocrinology |
| title | Dopamine receptor repertoire of human granulosa cells |
| title_full | Dopamine receptor repertoire of human granulosa cells |
| title_fullStr | Dopamine receptor repertoire of human granulosa cells |
| title_full_unstemmed | Dopamine receptor repertoire of human granulosa cells |
| title_short | Dopamine receptor repertoire of human granulosa cells |
| title_sort | dopamine receptor repertoire of human granulosa cells |
| url | http://www.rbej.com/content/5/1/40 |
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