. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. ORGAN CULTURE AND INJECTION OF NEURONS 47. 0 5 10 15 20 25 min. after addition of fluorescein di-(B-D-galactopyranoside) Figure 6. |3-galactosidase activity in a lobster neuron maintained 10 days in organ culture alter injection with LacZ cRNA. /3-galactosidase activity was detected in a living injected cell by addition of a fluorogenic substrate, fluorescein di-(fJ-D-galactopyranoside). (Left) The cell was photographed, using a confocal fluorescent microscope, at various time points (in minutes) following addition of the su
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. ORGAN CULTURE AND INJECTION OF NEURONS 47. 0 5 10 15 20 25 min. after addition of fluorescein di-(B-D-galactopyranoside) Figure 6. |3-galactosidase activity in a lobster neuron maintained 10 days in organ culture alter injection with LacZ cRNA. /3-galactosidase activity was detected in a living injected cell by addition of a fluorogenic substrate, fluorescein di-(fJ-D-galactopyranoside). (Left) The cell was photographed, using a confocal fluorescent microscope, at various time points (in minutes) following addition of the substrate. (Right) The intensity of fluorescence increased almost linearly Future uses of the organ culture method Long-term organ culture offers a controlled environment in which to perform experiments that were difficult to carry out in short-term studies using isolated nerve cords. This should allow us to explore the consequences of applying test sub- stances to central ganglia in more biologically relevant time frames. For example, the lobster molting hormone, 20-hy- droxyecdysone, is likely to have actions at both membrane and genomic levels (see Zakon, 1998). The genomic effects may take days to bring about observable changes at synaptic or circuit levels. The organ culture system offers enough time for such changes to be seen, in an environment in which tissues will continue to synthesize the RNA and protein required to trigger the changes. Long-term drug effects relating to amin- ergic function also can be examined in the cultured ganglia. At present we are particularly interested in chronic exposure of ganglia to Prozac (fluoxetine). In behavioral studies, acute exposure to Prozac has little effect on agonistic behavior in lobsters (Huber el ai, 1997); in contrast, chronic exposure increases the amount of time that animals are willing to fight (A. Delago, unpubl. obs.). Another exciting application of the organ culture method is in the experimental manipulation of gene e
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Keywords: ., bookauthorlilliefrankrat, booksubjectbiology, booksubjectzoology
