An improved method for in vivo electroporation of morpholinos into the adult zebrafish retina.
- Author:
Hai-Tao HOU
1
;
Jin-Yang LV
2
;
Zhi-Qiang ZHANG
1
;
Ying LU
3
;
Cui-Ping ZHOU
3
;
Tian-Qiu ZHOU
3
;
Shu-Qiang ZHANG
4
;
Hui XU
5
Author Information
1. Co-innovation Center of Neuroregeneration, Ministry of Education and Jiangsu Key Laboratory of Neuroregeneration, Nantong University, Nantong 226001, China.
2. Medical School, Nantong University, Nantong 226001, China.
3. Department of Ophthalmology, Affiliated Hospital, Nantong University, Nantong 226001, China.
4. Co-innovation Center of Neuroregeneration, Ministry of Education and Jiangsu Key Laboratory of Neuroregeneration, Nantong University, Nantong 226001, China. sqzhang0328@ntu.edu.cn.
5. Department of Ophthalmology, Affiliated Hospital, Nantong University, Nantong 226001, China. huixu82@126.com.
- Publication Type:Journal Article
- From:
Acta Physiologica Sinica
2018;70(1):47-51
- CountryChina
- Language:Chinese
-
Abstract:
In vivo electroporation of morpholinos (MOs) into the retina of adult zebrafish is an efficient method to study gene function related to retinal disease and regeneration. However, the currently reported methods are complicated with low MO transfer efficiency and high probability to cause collateral damage. The present study was aimed to optimize the existing MO electroporation methods. Two major changes were made to MO electroporation procedure in zebrafish retina. One was to coat the inner side of the electrode with ultrasonic gel. The other was to replace the commonly used round electrode with novel rectangular one. The results showed that the use of ultrasonic gel reduced collateral damage caused by retinal electroporation and simplified the experimental procedure. The rectangular electrode significantly increased transfection efficiency of MO electroporation. In particular, knocking down the expression of Ascl1a in the retina by using our method significantly inhibited the generation of retinal progenitor cells. These results suggest our method is the optimization of the current MO electroporation methods and may be a better alternative for relevant researchers.
