Ultrastructural dynamics of exocytosis in the ovulation-neurohormone producing caudo-dorsal cells of the freshwater snail Lymnaea stagnalis (L.)

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Abstract
The ultrastructural dynamics of exocytosis in the ovulation-stimulating neurosecretory Caudo-Dorsal Cells (CDC) of the freshwater snail L. stagnalis were studied after incubation of cerebral ganglia in Ringer's solutions with different concentrations of K+ and Ca2+. Detection of exocytosis was facilitated by the use of the tannic acid-glutaraldehyde fixation method (TAGO-method). In control Ringer (low K+) the frequency of exocytosis was rather low. Exocytosis mainly occurred as “terminal” exocytosis (TE); “intercellular” (ICE) and, particularly, “multiple” exocytosis (ME) took place infrequently. Incubation in high K+-containing Ringer strongly increased exocytotic activity. Compared to the controls the total frequency of exocytosis was 50x as high, whereas TE, ICE and ME occurred 6 x, 47x, and more than 300x as frequently, respectively. In high K+/Ca2+-free Ringer the total frequency of exocytosis was only 2x as high as in control Ringer. It is concluded that TE, ICE, and ME are normal, Ca2+-dependent exocytotic phenomena. The significance of their dynamics in response to K+-stimulation is discussed. The extremely high frequency of exocytosis, as well as the presence of “unaltered granule contents in transit”, is explained by assuming that an exocytotic event in the CDC lasts rather long, viz. some minutes. The results may reflect the physiological mechanism by which the CDC release their ovulation hormone. The possible involvement of “clear” and “large” electron lucent vesicles in membrane reuptake after exocytosis is considered.