Toxin of the marine alga Prymnesium patelliferum enhances voltage dependent Ca²⁺‐currents, elevates the cytosolic Ca²⁺‐concentration and facilitates hormone release in clonal rat pituitary cells

Abstract

The marine flagellate Prymnesium patelliferum produces toxins lethal to fish. The toxin extracted from the alga has haemolytic, cytotoxic and neurotoxic effects, but the action mechanisms of the toxin are not known in detail. We have examined the toxin effects on the voltage sensitive Ca(2+)-currents, the cytosolic Ca(2+)-level ([Ca2+]i) and the prolactin release in clonal rat anterior pituitary GH4C1 cells, which possess T- and L-type Ca(2+)-channels. The trans-membrane Ca(2+)-current was recorded using whole-cell voltage clamp. After 5-15 min exposure to the algal toxin at a final concentration of 50,000-100,000 cells mL-1, the Ca(2+)-currents through both the T- and L-channels showed a 2-3-fold enhancement. The voltage sensitivity of the Ca(2+)-currents was not affected by the algal toxin, and the toxin-induced currents were inhibited by 100 microM of the Ca(2+)-channel blocker D-600. In toxin-exposed cells microfluorometric measurements based on fura-2 revealed an increase of [Ca2+]i from 100-150 to 300-500 nM. This elevation was delayed and partially inhibited by 100 microM D-600. The algal toxin induced prolactin release in a dose-dependent manner, and this effect was inhibited by the Ca(2+)-channel blocker verapamil. We therefore conclude that the toxin of P. patelliferum affects the Ca2+ homeostasis of the pituitary cells by increasing the leak through voltage sensitive Ca(2+)-channels, resulting in increased [Ca2+]i and secretion of prolactin.

A TOXIC EXTRACT OF THE MARINE PHYTOFLAGELLATEPRYMNESIUM PARVUMINDUCES CALCIUM-DEPENDENT RELEASE OF GLUTAMATE FROM RAT BRAIN SYNAPTOSOMES

“…They also showed that the toxin increases the Ca 2+ and Na + influx, through the plasma membrane of synaptosomes. It is also reported that prymnesium toxin induces Ca 2+ influx in other cell preparations, such as rat pituary and glioma cells (Kolderup et al, 1995;Igarashi et al, 1998). These findings indicate that one could expect to find toxin-induced release of neurotransmitters.…”

“…This was attributed to an effect on the Na + gradient, diminishing the driving force for uptake. Later it was shown that the algal extract of prymnesium also could induce influx of Ca 2+ into both synaptosomes and rat pituitary cells (Kolderup et al, 1995;Mehldal et al, 1994). These findings indicate that the inhibitory effect of the toxin on glutamate uptake also may be a result of spontaneous release of neurotransmitters.…”