Transgenic mouse lines expressing synaptopHluorin in hippocampus and cerebellar cortex.
We generated six transgenic mouse lines in which synaptopHluorin (SpH), one of green fluorescent protein-based sensors of vesicular exocytosis, was expressed under the control of neuron-specific Thy-1.2 promoter. In situ hybridization study revealed that SpH mRNA was expressed in a broad spectrum of brain regions in four of them, whereas in others it was expressed in the specific regions of the hippocampus. In one particular line, SpH immunoreactivity was specifically observed in the mossy fiber presynaptic terminals of both hippocampus and cerebellar cortex. The fluorescence intensity of these presynaptic terminals was somewhat decreased by acidic buffer superfusion and greatly increased by vesicular neutralization of pH, indicating that the SpH molecules are mainly distributed in the synaptic vesicles. The exocytosis-dependent fluorescence increment was measured upon activation of a single presynaptic terminal. These transgenic lines are expected to facilitate morphological and physiological studies of presynaptic terminals in a variety of regions of the brain.
Araki
Rikita
R
Department of Developmental Biology and Neurosciences, Tohoku University Graduate School of Life Sciences, and CREST, JST, Sendai, Japan.
Sakagami
Hiroyuki
H
Yanagawa
Yuchio
Y
Hikima
Takuya
T
Ishizuka
Toru
T
Yawo
Hiromu
H
D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins; D002490:Central Nervous System / Q000378:metabolism; D002525:Cerebellar Cortex / Q000378:metabolism*; D003001:Cloning, Molecular; D005453:Fluorescence; D015870:Gene Expression; D049452:Green Fluorescent Proteins / Q000032:analysis* / Q000235:genetics; D006624:Hippocampus / Q000378:metabolism*; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D049408:Luminescent Agents / Q000032:analysis*; D008565:Membrane Proteins / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D018613:Microscopy, Confocal; D017729:Presynaptic Terminals / Q000737:chemistry*; D050683:R-SNARE Proteins; D012333:RNA, Messenger / Q000378:metabolism; D013572:Synaptic Vesicles / Q000737:chemistry; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
D051792:Basic Helix-Loop-Helix Proteins; D049452:Green Fluorescent Proteins; D049408:Luminescent Agents; D008565:Membrane Proteins; D050683:R-SNARE Proteins; D012333:RNA, Messenger; D014157:Transcription Factors; C095572:Hand1 protein, mouse