. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. EXPRESSION OF FARPs IN HELIX NEURONS 115 antibody to digoxigenin (Boehringer-Mannheim) for 2 h. The antibody was diluted 1 in 100 in phosphate buffered saline containing Tween 20 (PBS/T) plus 2% bovine serum albumin. The preparations were then washed ex- tensively in PBS/T and mounted in glycerol/PBS (9:1) for viewing on a scanning confocal-laser microscope (Bio- Rad MRC 600). N-propyl gallate (Sigma) was added to the PBS ( g/10 ml) as an antifade agent. Results Survey of Single and Clusters oflmmunoreactive Neurons
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. EXPRESSION OF FARPs IN HELIX NEURONS 115 antibody to digoxigenin (Boehringer-Mannheim) for 2 h. The antibody was diluted 1 in 100 in phosphate buffered saline containing Tween 20 (PBS/T) plus 2% bovine serum albumin. The preparations were then washed ex- tensively in PBS/T and mounted in glycerol/PBS (9:1) for viewing on a scanning confocal-laser microscope (Bio- Rad MRC 600). N-propyl gallate (Sigma) was added to the PBS ( g/10 ml) as an antifade agent. Results Survey of Single and Clusters oflmmunoreactive Neurons detected Histochemically Refer to Figure 1 for locations of neurons used and Table I for a summary of the results. Cerebral ganglia As a test of the dissection and RIA procedures, exper- iments were first made on the Cl neuron, which contains 5-hydroxytryptamine but does not stain positively with FMRFamide antisera. The level of immunoreactivity in this neuron was lower than the limit of detection, less than pmole/cell (see Table I). This result suggests that general contamination of samples during cell isolation did not pose a problem. C3 neuron. Previous histochemical and RIA data sug- gest that this neuron contains one or more FaRPs (Cottrell el ai. 1982). and most probably only FMRFamide and FLRFamide (Bewick el 1990). Assays of single C3 cells in the present study indicate levels of FMRFamide equivalent to about pmole/cell. Right mesocerebral cluster. The experiments of Elekes and Nassel (1990) showed that a mass of cells in the me- socerebral area of the right cerebral ganglion were stained with the FMRFamide antiserum. Because they are so abundant, we subjected extracts of these cells to HPLC before RIA (see data below). Pleura! ganglia Neurons equivalent to LPL1 and RPLI. Two large cells, which have the same location as LPL1 and RPL1 detected immunocytochemically in H. pomatia by Elekes and Nassel (1990), were isolated from H. aspersa pleural gan- g
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Keywords: ., bookauthorlilliefrankrat, booksubjectbiology, booksubjectzoology
