PMID- 29708533
OWN - NLM
STAT- MEDLINE
DCOM- 20180730
LR  - 20200416
IS  - 1940-087X (Electronic)
IS  - 1940-087X (Linking)
IP  - 134
DP  - 2018 Apr 16
TI  - A Simple and Efficient Method for In Vivo Cardiac-specific Gene Manipulation by 
      Intramyocardial Injection in Mice.
LID - 10.3791/57074 [doi]
LID - 57074
AB  - Gene manipulation specifically in the heart significantly potentiate the 
      investigation of cardiac disease pathomechanisms and their therapeutic potential. 
      In vivo cardiac-specific gene delivery is commonly achieved by either systemic or 
      local delivery. Systemic injection via tail vein is easy and efficient in 
      manipulating cardiac gene expression by using recombinant adeno-associated virus 
      9 (AAV9). However, this method requires a relatively high amount of vector for 
      efficient transduction, and may result in nontarget organ gene transduction. 
      Here, we describe a simple, efficient, and time-saving method of intramyocardial 
      injection for in vivo cardiac-specific gene manipulation in mice. Under 
      anesthesia (without ventilation), the pectoral major and minor muscles were 
      bluntly dissected, and the mouse heart was quickly exposed by manual 
      externalization through a small incision at the fourth intercostal space. 
      Subsequently, adenovirus encoding luciferase (Luc) and vitamin D receptor (VDR), 
      or short hairpin RNA (shRNA) targeting VDR, was injected with a Hamilton syringe 
      into the myocardium. Subsequent in vivo imaging demonstrated that luciferase was 
      successfully overexpressed specifically in the heart. Moreover, Western blot 
      analysis confirmed the successful overexpression or silencing of VDR in the mouse 
      heart. Once mastered, this technique can be used for gene manipulation, as well 
      as injection of cells or other materials such as nanogels in the mouse heart.
FAU - Fu, Yanan
AU  - Fu Y
AD  - Department of Cardiology, The First Affiliated Hospital of Bengbu Medical 
      College; Department of Cardiology, Ren Ji Hospital, School of Medicine, Shanghai 
      Jiao Tong University.
FAU - Jiang, Wenlong
AU  - Jiang W
AD  - Department of Cardiology, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong 
      University.
FAU - Zhao, Yichao
AU  - Zhao Y
AD  - Department of Cardiology, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong 
      University.
FAU - Huang, Yuli
AU  - Huang Y
AD  - Department of Cardiology, The First Affiliated Hospital of Bengbu Medical 
      College.
FAU - Zhang, Heng
AU  - Zhang H
AD  - Department of Cardiology, The First Affiliated Hospital of Bengbu Medical 
      College; 13965270077@139.com.
FAU - Wang, Hongju
AU  - Wang H
AD  - Department of Cardiology, The First Affiliated Hospital of Bengbu Medical 
      College.
FAU - Pu, Jun
AU  - Pu J
AD  - Department of Cardiology, The First Affiliated Hospital of Bengbu Medical 
      College; Department of Cardiology, Ren Ji Hospital, School of Medicine, Shanghai 
      Jiao Tong University; pujun310@hotmail.com.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Video-Audio Media
DEP - 20180416
PL  - United States
TA  - J Vis Exp
JT  - Journal of visualized experiments : JoVE
JID - 101313252
SB  - IM
MH  - Animals
MH  - Gene Transfer Techniques/*statistics & numerical data
MH  - Genetic Therapy/*methods
MH  - Genetic Vectors/pharmacology/*therapeutic use
MH  - Injections/*methods
MH  - Mice
MH  - Myocardium/cytology/*metabolism
PMC - PMC5933510
EDAT- 2018/05/01 06:00
MHDA- 2018/07/31 06:00
PMCR- 2020/04/16
CRDT- 2018/05/01 06:00
PHST- 2018/05/01 06:00 [entrez]
PHST- 2018/05/01 06:00 [pubmed]
PHST- 2018/07/31 06:00 [medline]
PHST- 2020/04/16 00:00 [pmc-release]
AID - 57074 [pii]
AID - 10.3791/57074 [doi]
PST - epublish
SO  - J Vis Exp. 2018 Apr 16;(134):57074. doi: 10.3791/57074.