Figure 5.
Suppression of the reg6 regenerating vessel phenotype by SKF91488 and histamine. (A) reg6 regenerating vessels formed swollen lumina 3 days post amputation (dpa) at 33°C (yellow arrows). (B) In the presence of 10 μM SKF91488, reg6 mutants developed significantly smaller and fewer swollen vessels (yellow arrows). Red arrows, amputation plane. (C) Quantitative data of the suppression of reg6 regenerating vessels by SKF91488 (n = 5 for each group). This experiment was repeated at least 3 times, and consistent results were observed. (D) Quantitative polymerase chain reaction (QPCR) analyses showed elevated levels of histamine methyltransferase (HMT) RNA in both reg6 2 and 3-dpa regenerates compared with the wild-type. (E) Both SKF91488 and histamine could reverse the reg6 vessel phenotypes. The number of swollen vessels in reg6 mutants was counted at 3 dpa at 33°C (blue bars; B, before treatment) before they were transferred to either fresh water (H2O), 10 μM SKF91488, or 1 mM histamine. The number of swollen vessels was counted again 12 h later (red bars; P, post-treatment). Shown is a representative set of data with 10 fish. Similar results were obtained from 2 independent experiments with different stocks of reg6 fish.