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Bryostatin Enhancement of Memory in Hermissenda

¹ Marine Biological Laboratory, Woods Hole, Massachusetts 02543
² Science Department, Roger Williams University, Bristol, Rhode Island 02809
³ Blanchette Rockefeller Neurosciences Institute, Johns Hopkins University, Rockville, Maryland 20850
⁴ Department of Biological Science and Technology, Tokai University, Shizuoka, Japan
⁵ Department of Biology, University of Michigan, Ann Arbor, Michigan 48104
⁶ Northeastern University, Boston, Massachusetts 02115

^* To whom correspondence should be addressed. E-mail: akuzirian{at}mbl.edu

Bryostatin, a potent agonist of protein kinase C (PKC), when administered to Hermissenda was found to affect acquisition of an associative learning paradigm. Low bryostatin concentrations (0.1 to 0.5 ng/ml) enhanced memory acquisition, while concentrations higher than 1.0 ng/ml down-regulated the pathway and no recall of the associative training was exhibited. The extent of enhancement depended upon the conditioning regime used and the memory stage normally fostered by that regime. The effects of two training events (TEs) with paired conditioned and unconditioned stimuli, which standardly evoked only short-term memory (STM) lasting 7 min, were—when bryostatin was added concurrently—enhanced to a long-term memory (LTM) that lasted about 20 h. The effects of both 4- and 6-paired TEs (which by themselves did not generate LTM), were also enhanced by bryostatin to induce a consolidated memory (CM) that lasted at least 5 days. The standard positive 9-TE regime typically produced a CM lasting at least 6 days. Low concentrations of bryostatin (<0.5 ng/ml) elicited no demonstrable enhancement of CM from 9-TEs. However, animals exposed to bryostatin concentrations higher than 1.0 ng/ml exhibited no behavioral learning.

Sharp-electrode intracellular recordings of type-B photoreceptors in the eyes from animals conditioned in vivo with bryostatin revealed changes in input resistance and an enhanced long-lasting depolarization (LLD) in response to light. Likewise, quantitative immunocytochemical measurements using an antibody specific for the PKC-activated Ca²⁺/GTP-binding protein calexcitin showed enhanced antibody labeling with bryostatin.

Animals exposed to the PKC inhibitor bisindolylmaleimide-XI (Ro-32-0432) administered by immersion prior to 9-TE conditioning showed no training-induced changes with or without bryostatin exposure. However, if animals received bryostatin before Ro-32, the enhanced acquisition and demonstrated recall still occurred. Therefore, pathways responsible for the enhancement effects induced by bryostatin were putatively mediated by PKC.

Overall, the data indicated that PKC activation occurred and calexcitin levels were raised during the acquisition phases of associative conditioning and memory initiation, and subsequently returned to baseline levels within 24 and 48 h, respectively. Therefore, the protracted recall measured by the testing regime used was probably due to bryostatin-induced changes during the acquisition and facilitated storage of memory, and not necessarily to enhanced recall of the stored memory when tested many days after training.

Abbreviations: CM, consolidated memory • CR, conditioned response • CS, conditioned stimulus • LLD, long-lasting depolarization • LTM, long-term memory • NSW, natural seawater • PKC, protein kinase C • Ro-32, Ro-32-0432, or bisindolylmaleimide-XI • RTE, random training event • STM, short-term memory • TE, training event • UCR, unconditioned response • US, unconditioned stimulus

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