PMID- 34324942
OWN - NLM
STAT- MEDLINE
DCOM- 20220204
LR  - 20220204
IS  - 1872-9754 (Electronic)
IS  - 0197-0186 (Linking)
VI  - 149
DP  - 2021 Oct
TI  - A novel method for detecting heteromeric complexes at synaptic level by combining 
      a modified method of proximity ligation assay with transmission electron 
      microscopy.
PG  - 105145
LID - S0197-0186(21)00191-1 [pii]
LID - 10.1016/j.neuint.2021.105145 [doi]
AB  - The heteromeric complexes of adenosine 2A receptor (A2AR) and 
      N-methyl-D-aspartate receptor (NMDAR) have recently been confirmed in cell 
      experiments, while its in situ detection at the subcellular level of brain tissue 
      has not yet been achieved. Proximity Ligation Assay (PLA) enables the detection 
      of low-abundance proteins and their interactions at the cellular level with high 
      specificity and sensitivity, while Transmission electron microscope (TEM) is an 
      excellent tool for observing subcellular structures. To develop a highly 
      efficient and reproducible technique for in situ detection of protein 
      interactions at subcellular levels, in this study, we modified the standard PLA 
      sample preparation method to make the samples suitable for analysis by 
      transmission electron microscopy. Using this technique, we successfully detected 
      the heteromers of A2AR and NMDAR1, the essential subunit of NMDA receptor on the 
      hippocampal synaptic structure in mice. Our results show that the distribution of 
      this heteromer is different in different hippocampal subregions. This technique 
      holds the potential for being a reliable method to detect protein interactions at 
      the subcellular level and unravel their unknown functions.
CI  - Copyright (c) 2021. Published by Elsevier Ltd.
FAU - Tan, Si-Wei
AU  - Tan SW
AD  - State Key Laboratory of Trauma, Burns and Combined Injury, Department of 
      Occupational Disease, Daping Hospital, Army Medical University, Chongqing, China.
FAU - Li, Ping
AU  - Li P
AD  - State Key Laboratory of Trauma, Burns and Combined Injury, Department of 
      Occupational Disease, Daping Hospital, Army Medical University, Chongqing, China.
FAU - Ning, Ya-Lei
AU  - Ning YL
AD  - State Key Laboratory of Trauma, Burns and Combined Injury, Department of 
      Occupational Disease, Daping Hospital, Army Medical University, Chongqing, China.
FAU - Zhao, Yan
AU  - Zhao Y
AD  - State Key Laboratory of Trauma, Burns and Combined Injury, Department of 
      Occupational Disease, Daping Hospital, Army Medical University, Chongqing, China.
FAU - Yang, Nan
AU  - Yang N
AD  - State Key Laboratory of Trauma, Burns and Combined Injury, Department of 
      Occupational Disease, Daping Hospital, Army Medical University, Chongqing, China.
FAU - Zhou, Yuan-Guo
AU  - Zhou YG
AD  - State Key Laboratory of Trauma, Burns and Combined Injury, Department of 
      Occupational Disease, Daping Hospital, Army Medical University, Chongqing, China. 
      Electronic address: yuanguo.zhou@gmail.com.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20210726
PL  - England
TA  - Neurochem Int
JT  - Neurochemistry international
JID - 8006959
RN  - 0 (NMDA receptor A1)
RN  - 0 (Receptor, Adenosine A2A)
RN  - 0 (Receptors, N-Methyl-D-Aspartate)
SB  - IM
MH  - Animals
MH  - Hippocampus/metabolism/*ultrastructure
MH  - Male
MH  - Mice
MH  - Mice, Inbred C57BL
MH  - Microscopy, Electron, Transmission/*methods
MH  - Protein Binding/physiology
MH  - Receptor, Adenosine A2A/metabolism/*ultrastructure
MH  - Receptors, N-Methyl-D-Aspartate/metabolism/*ultrastructure
MH  - Synapses/metabolism/*ultrastructure
OTO - NOTNLM
OT  - A2AR
OT  - NMDAR
OT  - PLA
OT  - PPI
OT  - Synapse
OT  - TEM
EDAT- 2021/07/30 06:00
MHDA- 2022/02/05 06:00
CRDT- 2021/07/29 20:12
PHST- 2021/01/06 00:00 [received]
PHST- 2021/07/22 00:00 [revised]
PHST- 2021/07/25 00:00 [accepted]
PHST- 2021/07/30 06:00 [pubmed]
PHST- 2022/02/05 06:00 [medline]
PHST- 2021/07/29 20:12 [entrez]
AID - S0197-0186(21)00191-1 [pii]
AID - 10.1016/j.neuint.2021.105145 [doi]
PST - ppublish
SO  - Neurochem Int. 2021 Oct;149:105145. doi: 10.1016/j.neuint.2021.105145. Epub 2021 
      Jul 26.