Figure 2
D-series resolvins RvD1 and aspirin-triggered RvD1 (AT-RvD1) reduce histamine (His)-stimulated increase in [Ca2+]i by preventing release of intracellular Ca2+ in conjunctival goblet cells. Result of time of addition of (a and b) RvD1 (10−8 M) or (c and d) AT-RvD1 (10−8 M) on histamine (10−5 M)-stimulated increase in [Ca2+]i in rat conjunctival goblet cells. Panels a and c are pseudo color images of [Ca2+]i in single goblet cells in response to histamine alone or with RvD1 or AT-RvD1 added at the same time as histamine or 50 s after addition of histamine. Intracellular Ca2+ response over time to histamine alone, RvD1, or AT-RvD1 added simultaneously with histamine or added 50 s after histamine is shown in b and d (top panels, from a representative experiment). Peak [Ca2+]i in response to histamine alone (open bars), RvD1, or AT-RvD1 added simultaneously with histamine (diagonal lines) or added at 60 s after histamine (cross-hatched lines) is shown in b and d lower panels from seven experiments. *P<0.05 (histamine vs. resolvin plus histamine). First arrow indicates addition of RvD1 with histamine. Second arrow indicates addition of RvD1 after histamine. (e) Use of capacitative Ca2+ store by RvD1 or AT-RvD1 in rat goblet cells. Left top panel indicates intracellular Ca2+ response over time in response to histamine (10−5 M) alone. Right top panel indicates thapsigargin (Thap; 10−5 M, first arrow) followed by histamine (10−5 M second arrow). A 30-min exposure with RvD1 (10−10 M) or AT-RvD1 (10−11 M) followed by thapsigargin (10−6 M, first arrow) and then histamine (10−5 M, second arrow) is indicated in the middle panels. Bottom panel indicates peak [Ca2+]i in response to histamine (10−5 M) alone; exposure of vehicle, RvD1, or AT-RvD1 for 30 min followed by thapsigargin (10−6 M) and then followed by histamine (10−5 M) from four experiments. #P<0.05 (histamine vs. resolvin plus thapsigargin plus histamine).
