Calcium (Ca$ sp{2+}$) and protein kinase C (PKC) are believed to act as intracellular signals triggering the activation of NADPH oxidase in neutrophils leading to superoxide generation. This was tested on bovine neutrophils by chelating extracellular and/or intracellular free Ca$ sp{2+}$ and by measuring PKC activity when the cells were stimulated by phorbol myristate acetate (PMA) or opsonized zymosan (OZ). Chelation of extracellular Ca$ sp{2+}$ with EGTA did not alter O$ sb2 sp{-}$ production from PMA stimulated cells. However, it did cause a 64% decrease in O$ sb2 sp{-}$ production in the neutrophils when stimulated with OZ. When intracellular Ca$ sp{2+}$ was chelated with BAPTA/AM, there was a significant decrease in O$ sb2 sp{-}$ generation following PMA activation. Yet, OZ activated cells, pre-treated with BAPTA/AM, showed an increase in the respiratory burst proportional to the chelator's concentration. Moreover, although OZ was previously shown to increase O$ sb2 sp{-}$ generation by neutrophils, no significant changes in PKC activity were observed. PMA stimulation led to an increase in PKC activity at the membrane level. Furthermore, treating the cells with calphostin C, a PKC activity inhibitor, caused a 69% decrease in O$ sb2 sp{-}$ production demonstrating the involvement of PKC in PMA-stimulated cells. However, no differences were observed between the OZ activated cells incubated with the inhibitor and the control cells. These data provide evidence that activation of NADPH oxidase can be achieved by either a PKC-dependent or a PKC-independent pathway depending on the stimulatory agent.
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:QMM.23983 |
| Date | January 1996 |
| Creators | Allard, Brenda. |
| Contributors | Zhao, Xin (advisor) |
| Publisher | McGill University |
| Source Sets | Library and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada |
| Language | English |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Format | application/pdf |
| Coverage | Master of Science (Department of Animal Science.) |
| Rights | All items in eScholarship@McGill are protected by copyright with all rights reserved unless otherwise indicated. |
| Relation | alephsysno: 001538789, proquestno: MM19792, Theses scanned by UMI/ProQuest. |
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