Presynaptic Homeostasis at CNS Nerve Terminals Compensates for Lack of a Key Ca2+ Entry Pathway

• Published in: Proceedings of the National Academy of Sciences - PNAS 2004-03, Vol.101 (10), p.3609-3614
• Main Authors: Piedras-Rentería, Erika S., Pyle, Jason L., Diehn, Max, Glickfeld, Lindsey L., Harata, Nobutoshi C., Cao, Yuqing, Kavalali, Ege T., Brown, Patrick O., Tsien, Richard W.
• Format: Article
• Language: English
• Subjects: Animals; Biological Sciences; Calcium Channels, N-Type - deficiency; Calcium Channels, N-Type - genetics; Calcium Signaling; Cells, Cultured; Complementary DNA; Cyclic AMP-Dependent Protein Kinases - metabolism; Deoxyribonucleic acid; DNA; Fluorescence; Gene expression; Gene Expression Profiling; Hippocampus; Hippocampus - cytology; Hippocampus - metabolism; Homeostasis; Messenger RNA; Mice; Mice, Knockout; Molecules; Nerves; Neurology; Neurons; Neurons - metabolism; Oligonucleotide Array Sequence Analysis; Presynaptic Terminals - metabolism; Ribonucleic acid; RNA; RNA, Messenger - genetics; RNA, Messenger - metabolism; Synapses
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.0308188100

At central synapses, P/Q-type Ca2+ channels normally provide a critical Ca2+ entry pathway for neurotransmission. Nevertheless, we found that nerve terminals lacking α 1 A( Ca v2.1), the pore-forming subunit of P/Q-type channels, displayed a remarkable preservation of synaptic function. Two consistent physiological changes reflective of synaptic homeostasis were observed in cultured hippocampal neurons derived from α 1 A (-/-) mice. First, the presynaptic response to an ionophore-mediated Ca2+ elevation was 50% greater, indicating an enhanced Ca2+ sensitivity of the release machinery. Second, basal miniature excitatory postsynaptic current frequency in α 1 A (-/-) neurons was increased 2-fold compared with WT neurons and occluded the normal response of presynaptic terminals to cAMP elevation, suggesting that the compensatory mechanism in α 1 A (-/-) synapses and the modulation of presynaptic function by PKA might share a final common pathway. We used cDNA microarray analysis to identify molecular changes underlying homeostatic regulation in the α 1 A (-/-) hippocampus. The 40,000 entries in our custom-made array included likely targets of presynaptic homeostasis, along with many other transcripts, allowing a wide-ranging examination of gene expression. The developmental pattern of changes in transcript levels relative to WT was striking; mRNAs at 5 and 11 days postnatal showed little deviation, but clear differences emerged by 22 days. Many of the transcripts that differed significantly in abundance corresponded to known genes that could be incorporated within a logical pattern consistent with the modulation of presynaptic function. Changes in endocytotic proteins, signal transduction kinases, and candidates for Ca2+-sensing molecules were consistent with implications of the direct physiological experiments.