D-aspartic acid in the nervous system of Aplysia limacina: Possible role in neurotransmission

In the marine mollusk Aplysia limacina, a substantial amount of endogenous D‐aspartic acid (D‐Asp) was found following its synthesis from L‐aspartate by an aspartate racemase. Concentrations of D‐Asp between 3.9 and 4.6 µmol/g tissue were found in the cerebral, abdominal, buccal, pleural, and pedal ganglia. In non nervous tissues, D‐Asp occurred at a very low concentration compared to the nervous system. Immunohistochemical studies conducted on cultured Aplysia neurons using an anti‐D‐aspartate antibody demonstrated that D‐Asp occurs in the soma, dendrites, and in synaptic varicosities. Synaptosomes and synaptic vesicles from cerebral ganglia were prepared and characterized by electron microscopy. HPLC analysis revealed high concentrations of D‐Asp together with L‐aspartate and L‐glutamate in isolated synaptosomes In addition, D‐Asp was released from synaptosomes by K+ depolarization or by ionomycin. D‐Asp was one of the principal amino acids present in synaptic vesicles representing about the 25% of total amino acids present in these cellular organelles. Injection of D‐Asp into live animals or addition to the incubation media of cultured neurons, caused an increase in cAMP content. Taken as a whole, these findings suggest a possible role of D‐Asp in neurotransmission in the nervous system of Aplysia limacina. © 2005 Wiley‐Liss, Inc.