Serotonin-induced cleavage of the atypical protein kinase C Apl III in Aplysia.
A constitutively active kinase, known as protein kinase Mζ (PKMζ), is proposed to act as a long-lasting molecular memory trace. While PKMζ is formed in rodents through translation of a transcript initiating in an intron of the protein kinase Cζ (PKCζ) gene, this transcript does not exist in Aplysia californica despite the fact that inhibitors of PKMζ erase memory in Aplysia in a fashion similar to rodents. We have previously shown that, in Aplysia, the ortholog of PKCζ, PKC Apl III, is cleaved by calpain to form a PKM after overexpression of PKC Apl III. We now show that kinase activity is required for this cleavage. We further use a FRET reporter to measure cleavage of PKC Apl III into PKM Apl III in live neurons using a stimulus that induces plasticity. Our results show that a 10 min application of serotonin induces cleavage of PKC Apl III in motor neuron processes in a calpain- and protein synthesis-dependent manner, but does not induce cleavage of PKC Apl III in sensory neuron processes. Furthermore, a dominant-negative PKM Apl III expressed in the motor neuron blocked the late phase of intermediate-term facilitation in sensory-motor neuron cocultures induced by 10 min of serotonin. In summary, we provide evidence that PKC Apl III is cleaved into PKM Apl III during memory formation, that the requirements for cleavage are the same as the requirements for the plasticity, and that PKM in the motor neuron is required for intermediate-term facilitation.
Department of Psychology, McGill University, Montreal Neurological Institute, Montreal, Quebec H3A 1B1, Canada.
Joanna K
JK
Bougie
Diancai
D
Cai
Margaret
M
Hastings
Carole A
CA
Farah
Shanping
S
Chen
Xiaotang
X
Fan
Patrick K
PK
McCamphill
David L
DL
Glanzman
Wayne S
WS
Sossin
D000818:Animals; D001048:Aplysia / Q000201:enzymology*; D002460:Cell Line; D002478:Cells, Cultured; D007527:Isoenzymes / Q000378:metabolism; D008568:Memory / Q000502:physiology*; D009046:Motor Neurons / Q000201:enzymology; D011493:Protein Kinase C / Q000378:metabolism*; D012701:Serotonin / Q000502:physiology*; D000099221:Protein Kinase C zeta
D007527:Isoenzymes; D011493:Protein Kinase C; D012701:Serotonin; D000099221:Protein Kinase C zeta