A new minimally-invasive method for microinjection into the mouse spinal dorsal horn

作者：Kohro Yuta; Sakaguchi Emi; Tashima Ryoichi; Tozaki Saitoh Hidetoshi; Okano Hideyuki; Inoue Kazuhide^*; Tsuda Makoto

来源：Scientific Reports, 2015, 5(1): 14306.

DOI：10.1038/srep14306

Noninvasive gene delivery to the spinal dorsal horn (SDH) remains challenging because existing methods to directly microinject vectors require laminectomy, which leads to tissue damage and inflammation. Such responses might hamper accurate readouts of cellular and behavioural effects of an introduced gene. Here we develop a new minimally-invasive SDH microinjection technique without the need of laminectomy in which a microcapillary is inserted into the SDH parenchyma through an intervertebral space. Using this method, we microinjected adeno-associated virus with an astrocytic promoter into the SDH and achieved efficient gene expression in an astrocyte-specific manner without gliosis, neuronal loss or inflammation. Furthermore, astrocytic loss-and gain-of-function of the transcription factor STAT3 by expressing a dominant-negative form and a constitutive-active form of STAT3, respectively, demonstrated the necessity and sufficiency of astrocytic STAT3 in the maintenance of neuropathic pain following peripheral nerve injury, a debilitating chronic pain state in which currently available treatments are frequently ineffective. Thus, our technique enables manipulation of gene expression in cell type-and spatial-specific manners without adverse effects, and may be useful for research in SDH physiology and pathology.

• 出版日期2015-9-21