. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. REPORTS FROM MBL GENERAL SCIENTIFIC MEETINGS 331 Long-Term Monitoring of Ca2+ in Cultured Aplysia Neurons with Fluorescent Probes Felix Strwnwasser, Joseph Mclntyre, and Carol A. Rainville (Marine Biological Laboratory) Long-term monitoring and imaging of free Ca:+ in cultured neurons and other cells, would be useful for a variety of purposes, particularly in observing the role of the ion in slow events such as growth and circadian oscillations. Ca2+ is essential to the ac- complishment of most cellular processes, including
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. REPORTS FROM MBL GENERAL SCIENTIFIC MEETINGS 331 Long-Term Monitoring of Ca2+ in Cultured Aplysia Neurons with Fluorescent Probes Felix Strwnwasser, Joseph Mclntyre, and Carol A. Rainville (Marine Biological Laboratory) Long-term monitoring and imaging of free Ca:+ in cultured neurons and other cells, would be useful for a variety of purposes, particularly in observing the role of the ion in slow events such as growth and circadian oscillations. Ca2+ is essential to the ac- complishment of most cellular processes, including movement, secretion, early events of fertilization, and growth. We describe in this report some results of long-term Ca:+-imaging in the bag cell (BC) neurons of Aplysia. The BC neurons synthesize and release egg-laying hormone, a peptide that controls egg laying (reviewed in 1). As neurons in primary culture, the neuropep- tidergic BCs regenerate their neurites and maintain their elec- trophysiological properties, as observed in the intact preparation (2). We describe our methods and preliminary observations on loading of the fluorescent Ca2+ probes, the development of Ca:+ hot zones and spots in the soma during neurite extension, spon- taneous oscillations of Ca:+ in the nucleus, and large spontaneous increases in Ca2+ in the cytoplasm and nucleus. Primary cultures of BC neurons from Aplysia californica were prepared by treating the abdominal ganglion with up to 11 units/ ml of neutral protease from Bacillus polymyxa (Sigma) for 3 h at 30°C, followed by rinses in the presence of BSA (24 mg/ml), and trituration onto 31mm glass cover slips. Artificial seawater (ASW) was used with glucose and penicillin or streptomycin, instead of either MEM or L-15 media, which had been used in the past (2). We use either fura-2, AM or fluo-3, AM (Molecular Probes) as the calcium indicators (3,4). Neurons are loaded for 30 to 45 min in 1 ml ASW containing 3 X 10~6 A/ indicator at 21°C; the lo
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Keywords: ., bookauthorlilliefrankrat, booksubjectbiology, booksubjectzoology
