. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. CARBONIC ANHVDRASE IN A SEA ANEMONE 499 The blots were washed in blocking buffer and incubated for 2 h in a 1:1000 dilution of the secondary antibody, an alkaline phosphatase-conjugated, donkey anti-sheep IgG (Sigma). In the development, nitro blue tetrazolium (NBT) and 5-bromo-4-chloro-3-indolyl phosphate (BCIP) were used as the substrates (Engvall, 1980; Johnstone and Thorpe, 1987). Results CA activity in symbiotic, aposymbiotic. and repopiilated anemones To determine whether CA activity in animal homog- enate is correlate
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. CARBONIC ANHVDRASE IN A SEA ANEMONE 499 The blots were washed in blocking buffer and incubated for 2 h in a 1:1000 dilution of the secondary antibody, an alkaline phosphatase-conjugated, donkey anti-sheep IgG (Sigma). In the development, nitro blue tetrazolium (NBT) and 5-bromo-4-chloro-3-indolyl phosphate (BCIP) were used as the substrates (Engvall, 1980; Johnstone and Thorpe, 1987). Results CA activity in symbiotic, aposymbiotic. and repopiilated anemones To determine whether CA activity in animal homog- enate is correlated with the presence of algae in animal tissue, CA activity was measured in animal tissue from (1) control symbiotic anemones, (2) aposymbiotic ane- mones, and (3) repopulated anemones. Both control symbiotic and repopulated symbiotic anemones were light brown and had similar average CA activities of ± and ± ApH units • min ' • mg protein ', respectively. In contrast, the aposymbiotic animals were white, almost transparent, and had a significantly lower average value of ± ApH units • min ' • mg pro- tein -' (Fig. 1). Change in CA activity with a change in numbers of algae To determine whether CA activity would change with a change in numbers of algae, anemones were sampled kinetically, as described above. The number of algae lost with increasing time in the dark was quantified in both cold shock and dark treated anemones (Fig. 2). After just. Control Aposymbiotic Repopulated Figure 1. CA activity in control anemones, anemones rendered apo- symbiotic by cold shock treatment and kept in the dark for 10 weeks, and anemones rendered aposymbiotic. kept in the dark, and subsequently reinfected with zooxanthellae. Each value is a mean ± SD of the mean (n = 5). * = different from aposymbiotic by P < .0001. ** = different from aposymbiotic by .0001 < P < .005 as calculated from a one way ANOVA. in, rl E in- IOJ i. * I 3 6 10 17 24 32 Days
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Keywords: ., bookauthorlilliefrankrat, booksubjectbiology, booksubjectzoology
