Intrinsic Expression of Coagulation Factors and Protease Activated Receptor 1 (PAR1) in Photoreceptors and Inner Retinal Layers
The aim of this study was to characterize the distribution of the thrombin receptor, protease activated receptor 1 (PAR1), in the neuroretina. Neuroretina samples of wild-type C57BL/6J and PAR1<sup>−/−</sup> mice were processed for indirect immunofluorescence and Western blot analysis. R...
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| Format: | Article |
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MDPI AG
2022-01-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/2/984 |
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| author | Zehavit Goldberg Ifat Sher Lamis Qassim Joab Chapman Ygal Rotenstreich Efrat Shavit-Stein |
| author_facet | Zehavit Goldberg Ifat Sher Lamis Qassim Joab Chapman Ygal Rotenstreich Efrat Shavit-Stein |
| author_sort | Zehavit Goldberg |
| collection | DOAJ |
| description | The aim of this study was to characterize the distribution of the thrombin receptor, protease activated receptor 1 (PAR1), in the neuroretina. Neuroretina samples of wild-type C57BL/6J and PAR1<sup>−/−</sup> mice were processed for indirect immunofluorescence and Western blot analysis. Reverse transcription quantitative real-time PCR (RT-qPCR) was used to determine mRNA expression of coagulation Factor X (FX), prothrombin (PT), and PAR1 in the isolated neuroretina. Thrombin activity following KCl depolarization was assessed in mouse neuroretinas ex vivo. PAR1 staining was observed in the retinal ganglion cells, inner nuclear layer cells, and photoreceptors in mouse retinal cross sections by indirect immunofluorescence. PAR1 co-localized with rhodopsin in rod outer segments but was not expressed in cone outer segments. Western blot analysis confirmed PAR1 expression in the neuroretina. Factor X, prothrombin, and PAR1 mRNA expression was detected in isolated neuroretinas. Thrombin activity was elevated by nearly four-fold in mouse neuroretinas following KCl depolarization (0.012 vs. 0.044 mu/mL, <i>p</i> = 0.0497). The intrinsic expression of coagulation factors in the isolated neuroretina together with a functional increase in thrombin activity following KCl depolarization may suggest a role for the PAR1/thrombin pathway in retinal function. |
| first_indexed | 2024-03-10T01:16:46Z |
| format | Article |
| id | doaj.art-19b736a59cec408b8b3dcd68a7170ce3 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T01:16:46Z |
| publishDate | 2022-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-19b736a59cec408b8b3dcd68a7170ce32023-11-23T14:07:42ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-01-0123298410.3390/ijms23020984Intrinsic Expression of Coagulation Factors and Protease Activated Receptor 1 (PAR1) in Photoreceptors and Inner Retinal LayersZehavit Goldberg0Ifat Sher1Lamis Qassim2Joab Chapman3Ygal Rotenstreich4Efrat Shavit-Stein5Goldschleger Eye Institute, Sheba Medical Center, Ramat Gan 5266202, IsraelGoldschleger Eye Institute, Sheba Medical Center, Ramat Gan 5266202, IsraelDepartment of Neurology, Sheba Medical Center, Ramat Gan 5266202, IsraelDepartment of Neurology, Sheba Medical Center, Ramat Gan 5266202, IsraelGoldschleger Eye Institute, Sheba Medical Center, Ramat Gan 5266202, IsraelDepartment of Neurology, Sheba Medical Center, Ramat Gan 5266202, IsraelThe aim of this study was to characterize the distribution of the thrombin receptor, protease activated receptor 1 (PAR1), in the neuroretina. Neuroretina samples of wild-type C57BL/6J and PAR1<sup>−/−</sup> mice were processed for indirect immunofluorescence and Western blot analysis. Reverse transcription quantitative real-time PCR (RT-qPCR) was used to determine mRNA expression of coagulation Factor X (FX), prothrombin (PT), and PAR1 in the isolated neuroretina. Thrombin activity following KCl depolarization was assessed in mouse neuroretinas ex vivo. PAR1 staining was observed in the retinal ganglion cells, inner nuclear layer cells, and photoreceptors in mouse retinal cross sections by indirect immunofluorescence. PAR1 co-localized with rhodopsin in rod outer segments but was not expressed in cone outer segments. Western blot analysis confirmed PAR1 expression in the neuroretina. Factor X, prothrombin, and PAR1 mRNA expression was detected in isolated neuroretinas. Thrombin activity was elevated by nearly four-fold in mouse neuroretinas following KCl depolarization (0.012 vs. 0.044 mu/mL, <i>p</i> = 0.0497). The intrinsic expression of coagulation factors in the isolated neuroretina together with a functional increase in thrombin activity following KCl depolarization may suggest a role for the PAR1/thrombin pathway in retinal function.https://www.mdpi.com/1422-0067/23/2/984thrombinPAR1neuroretinarodsconesphotoreceptor |
| spellingShingle | Zehavit Goldberg Ifat Sher Lamis Qassim Joab Chapman Ygal Rotenstreich Efrat Shavit-Stein Intrinsic Expression of Coagulation Factors and Protease Activated Receptor 1 (PAR1) in Photoreceptors and Inner Retinal Layers International Journal of Molecular Sciences thrombin PAR1 neuroretina rods cones photoreceptor |
| title | Intrinsic Expression of Coagulation Factors and Protease Activated Receptor 1 (PAR1) in Photoreceptors and Inner Retinal Layers |
| title_full | Intrinsic Expression of Coagulation Factors and Protease Activated Receptor 1 (PAR1) in Photoreceptors and Inner Retinal Layers |
| title_fullStr | Intrinsic Expression of Coagulation Factors and Protease Activated Receptor 1 (PAR1) in Photoreceptors and Inner Retinal Layers |
| title_full_unstemmed | Intrinsic Expression of Coagulation Factors and Protease Activated Receptor 1 (PAR1) in Photoreceptors and Inner Retinal Layers |
| title_short | Intrinsic Expression of Coagulation Factors and Protease Activated Receptor 1 (PAR1) in Photoreceptors and Inner Retinal Layers |
| title_sort | intrinsic expression of coagulation factors and protease activated receptor 1 par1 in photoreceptors and inner retinal layers |
| topic | thrombin PAR1 neuroretina rods cones photoreceptor |
| url | https://www.mdpi.com/1422-0067/23/2/984 |
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