Cellular and Molecular Neurobiology. 2020 Oct 15
Hui-Ling Tang, Si-Yu Chen, Huan Zhang, Ping Lu, Wei-Wen Sun, Mei-Mei Gao, Xiang-Da Zeng, Tao Su & Yue-Sheng Long
Products used in the paper Details Operation
AAV vector pAAV2-ALOXE3-mCherry The AAV vector pAAV2-ALOXE3-mCherry containing the coding sequences of mouse Aloxe3 (GenBank accession ID: NM_011786) and red fluorescent protein mCherry was provided by PackGene Biotech (Guangzhou, China). Request Quote

Research Field: CNS

Keywords: Arachidonate lipoxygenase 3, Spatiotemporal expression, Arachidonic acid, Neural development, Seizure

AAV Serotype: AAV2

Dose: The virus particles (2 μl, 1 × 10^8 GC/ml) were injected bilaterally into the hippocampal CA3 region (2.7 mm behind bregma, 3.2 mm lateral, and 2.7 mm below the dura) using a Hamilton syringe (Cat#: 80000, Hamilton) at a speed of 0.3 μl/min.

Targeted organ: brain

Animal or cell line strain: Male Swiss mice


Arachidonic acid (AA), a polyunsaturated fatty acid, is involved in the modulation of neuronal excitability in the brain. Arachidonate lipoxygenase 3 (ALOXE3), a critical enzyme in the AA metabolic pathway, catalyzes the derivate of AA into hepoxilins. However, the expression pattern of ALOXE3 and its role in the brain has not been described until now. Here we showed that the levels of Aloxe3 mRNA and protein kept increasing since birth and reached the highest level at postnatal day 30 in the mouse hippocampus and temporal cortex. Histomorphological analyses indicated that ALOXE3 was enriched in adult hippocampus, somatosensory cortex and striatum. The distribution was restricted to the neurites of function-specific subregions, such as mossy fibre connecting hilus and CA3 neurons, termini of Schaffer collateral projections, and the layers III and IV of somatosensory cortex. The spatiotemporal expression pattern of ALOXE3 suggests its potential role in the modulation of neural excitability and seizure susceptibility. In fact, decreased expression of ALOXE3 and elevated concentration of AA in the hippocampus was found after status epilepticus (SE) induced by pilocarpine. Local overexpression of ALOXE3 via adeno-associated virus gene transfer restored the elevated AA level induced by SE, alleviated seizure severities by increasing the latencies to myclonic switch, clonic convulsions and tonic hindlimb extensions, and decreased the mortality rate in the pilocarpine-induced SE model. These results suggest that the expression of ALOXE3 is a crucial regulator of AA metabolism in brain, and potentially acts as a regulator of neural excitability, thereby controlling brain development and seizure susceptibility.

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