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Intracellular free-calcium concentration ([Ca2+]i) was measured in lamprey spinal axons using the fluorescent calcium indicator fura 2. We used both a photomultiplier tube and a video-image processing system to measure the temporal and spatial distributions of [Ca2+]i in the proximal segments of transected axons. Within 3 min following transection, a spatially graded increase in the [Ca2+]i was apparent in the last few millimeters of the axons. Superimposed on the initial gradient was a moving front of calcium that progressed up the axon, reaching 1.6 mm from the cut end in 3 hr. The [Ca2+]i behind the moving front exceeded 10 microM. This movement of Ca2+ was greatly reduced by an externally applied electrical field with the cathode distal to the lesion and was increased by an applied field of the opposite polarity. When axons were transected in Ca2(+)-free medium, no increases in [Ca2+]i occurred. One d after transection, [Ca2+]i was at or below the precut levels, except in the distal 250 microns, where it remained slightly elevated. Therefore, axons can survive the high levels of [Ca2+]i that occur after transection and can reestablish normal [Ca2+]i levels within 24 hr. Measurements of both the diffusion coefficient and the fluorescence polarization of fura 2 indicate that the dye is not significantly bound to axoplasmic components.

We have exposed cultures of PC12 cells to uniform DC electric fields following the addition of NGF. The success of these experiments relied upon the design of new chambers enabling fields to be applied to mammalian cell cultures. After 48 h of field application, the distribution of neurite outgrowths was biased towards the anode. More neurites faced the anode than would be expected if growth was uniform. The magnitude of this bias was strongly correlated with field strength, with a threshold value of about 1 mV/mm. At field strengths above 30 mV/mm, the neurites growing towards the cathode were shorter than those growing towards the anode or perpendicular to the field. This response was not correlated with field strength. This report confirms that mammalian neurons respond to electrical fields and supports the notion that neurites are influenced by endogenous electrical fields during development. As far as we are aware, this is the only report that documents a response towards the anode.

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Attempts have been made to use the fluorescent calcium quin-2 to measure cytoplasmic free calcium in a variety of plant cells. Failure to measure intracellular fluorescence can be attributed to extracellular hydrolysis of the membranepermeable ester quin-2-AM used to load the cells. Attempts to overcome this problem by long incubation times showed that the by-product of ester hydrolysis, formaldehyde, is inhibitory to cell growth. Thus, it seems that the applicability of the acetoxymethyl ester of quin-2 for cytoplasmic calcium measurements in plant cells is limited, though quin-2 could still be very useful in some cells, especially if more suitable esters or micro-injection are used.

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Although much information about such processes as cell cycle control, second messenger systems, protein kinases and steroid hormone action has been collected from studies of Xenopus oocyte maturation, we still have very little idea about how the steroid hormone, progesterone, signals the resumption of meiosis from the oocyte plasma membrane. In this review we re-examine the data on second messenger systems in Xenopus oocytes and discuss some of the unresolved questions about hormone signal transduction during maturation. We outline some reasons for the contradictions in the literature and offer some suggestions for avenues of future research.