Complementing Confocal Detection of Antibody‐labeled Lysophosphatidic Acid Receptors in Human Gingivae with Label‐free Second Harmonic Generation Confocal Microscopy Detection of Collagen
| Autor: | Afolabi O. Ogunleye, D. Roselyn Cerutis, Michael G. Nichols, Shakeel Khan, Michael Hironaka, Timothy P. McVaney, Takanari Miyamoto |
|---|---|
| Rok vydání: | 2015 |
| Předmět: |
biology
medicine.diagnostic_test Chemistry Confocal Second-harmonic generation Fibril Immunofluorescence Biochemistry law.invention Cell biology chemistry.chemical_compound Confocal microscopy law Lysophosphatidic acid Genetics biology.protein medicine Antibody Receptor Molecular Biology Biotechnology |
| Zdroj: | The FASEB Journal. 29 |
| ISSN: | 1530-6860 0892-6638 |
| DOI: | 10.1096/fasebj.29.1_supplement.lb26 |
| Popis: | We have found that lysophosphatidic acid (LPA) controls the healing and [Ca2+]i responses of human gingival fibroblasts via the LPA1 and LPA3 receptor subtypes, and that LPA3 may modulate their inflammatory responses. We have used immunofluorescence to detect LPA receptor (LPAR) distribution in human gingivae, but that technique does not afford visualization of the state of collagen fiber organization. We hypothesized that using the second harmonic generation (SHG) technique would enable detection of changes in the appearance, collagen fibril density, and fibril organizational structure of human gingiva, and that this technique would complement visualizing LPAR expression by confocal microscopy. Pieces of intact, paraformaldehyde-fixed, methanol:acetone (1:1) de-lipidated attached gingivae [~1.25 mm thick (from IRB-approved, consented donors) normal or with moderate-severe periodontal disease] were used. Confocal analysis using anti-LPA1 and anti–LPA3 antibodies confirmed that LPA3 is primarily found in t... |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Plný text