cFos-ANAB: A cFos-based Web Tool for Exploring Activated Neurons and Associated Behaviors / 神经科学通报·英文版

cFos is one of the most widely-studied genes in the field of neuroscience. Currently, there is no systematic database focusing on cFos in neuroscience. We developed a curated database-cFos-ANAB-a cFos-based web tool for exploring activated neurons and associated behaviors in rats and mice, comprising 398 brain nuclei and sub-nuclei, and five associated behaviors: pain, fear, feeding, aggression, and sexual behavior. Direct relationships among behaviors and nuclei (even cell types) under specific stimulating conditions were constructed based on cFos expression profiles extracted from original publications. Moreover, overlapping nuclei and sub-nuclei with potentially complex functions among different associated behaviors were emphasized, leading to results serving as important clues to the development of valid hypotheses for exploring as yet unknown circuits. Using the analysis function of cFos-ANAB, multi-layered pictures of networks and their relationships can quickly be explored depending on users' purposes. These features provide a useful tool and good reference for early exploration in neuroscience. The cFos-ANAB database is available at www.cfos-db.net .

The roles of GluN3-containing N-methyl-D-aspartate receptor in central nerve system / 浙江大学学报·医学版

The N-methyl-D-aspartate receptor (NMDAR) in central nerve system is mostly composed of GluN1 and GluN2 subunits. The classical NMDAR has been intensively studied. However, GluN3‑containing NMDAR is much less expressed and have atypical channel properties. Recently, accumulating evidences have revealed two types of GluN3‑containing NMDAR: glutamate-gated GluN1/GluN2/GluN3 NMDAR and glycine-gated GluN1/GluN3 NMDAR. The former may play important roles in regulating synapse maturation and pruning non-used synapses, and its elevated expression at the adult stage may alter synaptic reorganization in some neuropsychiatric disorders. The latter is expressed in the medial habenula and involves in control of aversion. This article reviews the recent progresses on the expression, functional properties of GluN3‑containing atypical NMDARs and the physiological and pathological relevance.

Clinical analysis on diagnosis and treatment of 116 patients with primary duodenal malignant tumor / 中国医师进修杂志

Objective:To explore the diagnosis, surgical methods and therapeutic effect of primary duodenal malignant tumor.Methods:The clinical data of 116 patients with primary duodenal malignant tumor from January 2010 to December 2018 were retrospectively analyzed.Results:Among 116 patients, adenocarcinoma was in 74 cases, interstitial tumor was in 25 cases, carcinoid was in 9 cases, the others was in 8 cases. Before operation, duodenoscopy was performed in 107 cases, and CT examination was performed in 76 cases. There were 57 cases of pancreaticoduodenectomy, 15 cases of duodenal segmental resection, 13 cases of subtotal gastrectomy and duodenal bulbar resection, 13 cases of duodenal partial resection, and 18 cases of palliative short circuit operation. The total incidence of postoperative complication was 31.9% (37/116), including pancreatic fistula in 8 cases (grade B 5 cases, grade C 3 cases), biliary fistula in 6 cases, abdominal infection in 5 cases, pulmonary infection in 4 cases, intestinal fistula in 3 cases, delayed gastric emptying in 3 cases, and hemorrhage in 8 cases. Four cases (3.4%) died during the perioperative period. Single factor Cox regression analysis result showed that the postoperative survival time was related to the tumor differentiation degree, operation method, tumor infiltration degree and lymphatic metastasis ( P<0.05 or <0.01); multi-factor Cox regression analysis results showed that the operation method, tumor infiltration degree and lymphatic metastasis were the independent risk factors for the postoperative survival time of patients with primary duodenal malignant tumor ( P<0.05). The patients were followed up until June 2021, and 9 cases were lost to follow-up. Kaplan-Meier survival curve analysis result showed that the postoperative overall 1-,3- and 5-year survival rates were 82.11%, 57.56% and 33.11%, respectively. Conclusions:Adenocarcinoma is the main primary malignant tumor of duodenum. Duodenoscopy and CT are the main examination methods. Radical resection is the most effective treatment for primary duodenal malignant tumor, and pancreaticoduodenectomy is the first choice. Surgical method, tumor infiltration degree and lymphatic metastasis are the independent risk factors affecting the prognosis of patients.

cFos-ANAB: A cFos-based Web Tool for Exploring Activated Neurons and Associated Behaviors / 神经科学通报·英文版

cFos is one of the most widely-studied genes in the field of neuroscience. Currently, there is no systematic database focusing on cFos in neuroscience. We developed a curated database—cFos-ANAB—a cFos-based web tool for exploring activated neurons and associated behaviors in rats and mice, comprising 398 brain nuclei and sub-nuclei, and five associated behaviors: pain, fear, feeding, aggression, and sexual behavior. Direct relationships among behaviors and nuclei (even cell types) under specific stimulating conditions were constructed based on cFos expression profiles extracted from original publications. Moreover, overlapping nuclei and sub-nuclei with potentially complex functions among different associated behaviors were emphasized, leading to results serving as important clues to the development of valid hypotheses for exploring as yet unknown circuits. Using the analysis function of cFos-ANAB, multi-layered pictures of networks and their relationships can quickly be explored depending on users’ purposes. These features provide a useful tool and good reference for early exploration in neuroscience. The cFos-ANAB database is available at www.cfos-db.net.

Advances on GABAergic interneurons in autism spectrum disorders / 浙江大学学报·医学版

More and more evidences support that the abnormality of GABAergic interneurons is associated with autism spectrum disorders (ASD), epilepsy, schizophrenia and other neurodevelopmental disorders. In recent years, numerous drugs have been developed to regulate ion channels and receptors in GABAergic interneurons, including sodium channels and N-methyl-D-aspartate (NMDA) receptors. The activators of Na channel can enhance the action potential of GABAergic interneurons by reducing the inactivation of Na channel. NMDA receptor, as a potential therapeutic target of ASD, can restore the NMDA function of GABAergic interneurons, which would be used to treat behavioral defects. In addition, there are many ion channels and receptors on GABAergic interneurons related to ASD. This article reviews GABAergic interneurons in the pathogenesis of ASD and the related interventions.

Clinical feature and gene detection in one case of Wiskott-Aldrich syndrome / 临床儿科杂志

Objective To explore the characteristics of WAS gene mutation in Wiskott-Aldrich syndrome (WAS). Methods The clinical data of one infant with WAS were retrospectively analyzed. All exons and flanking sequences in WAS gene were detected by PCR. Results A 5-month-old boy, who has a history of eczema and recurrent infection, was hospitalized for thrombocytopenia. CD8+ and CD4+T cell were increased while CD19+B cell was normal. Bone marrow cytology suggested megakaryocyte mature hindrance. WAS gene detection found C.880 A?>?G (p.Ile294Val) mutation, but no mutations were found in parents. This site was a pathogenic mutation predicted by Polyphen 2 software and SIFT software. Besides, sequence conservation analysis of different species found it was a conservative site and structural prediction analysis revealed it may affect the normal protein structure. This site of mutation has not been reported before. Conclusions Gene detection can make early diagnosis of WAS and C.880 A?>?G (p.Ile294Val) is a new mutation.

Expressions of Wnt2 and β-catenin in Doxorubicin-induced myocardial injury and their relationships with p53 / 中华实用儿科临床杂志

Objective To investigate the expressions of Wnt2 and β-catenin in Doxorubicin (DOX)-induced myocardial injury and to explore their roles in myocardial cell apoptosis.Methods Cardiomyoblast cells were damaged by different concentrations of DOX(1 mg/L,2 mg/L,3 mg/L,4 mg/L) for 72 h.The effect of different concentrations of DOX on cardiomyocyte growth curve was detected according to the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-h-tetrazolium bromide(MTT) assay.DOX(1 mg/L) was used to induce the model of cardiomyoblast cell injury.Cardiomyocytes were divided into 4 groups:group A:DOX-injured cardiomyocytes for 12 h ;group B:DOX-injured cardiomyocytes for 24 h ; group C:DOX-injured cardiomyocytes for 48 h; group D:normal cardiomyocytes.The expressions of Wnt2,β-catenin and p53 were observed by Western blot and reverse transcription polymerase chain reaction(RT-PCR) at the time point of 12 h,24 h and 48 h.Results DOX significantly inhibited cardiomyocyte proliferation in a dose dependent fashion.The protein and mRNA expressions of Wnt2 increased in the DOX-induced myocardial injury group compared with the group D,with statistical significance (F =224.115,P ＜ 0.05) ;The expressions of β-catenin,p53 were significantly increased compared with the group D,and the higher expression appeared with the time extending(F =188.145,231.927,all P ＜ 0.05).Significantly positive correlation between Wnt2 and β-catenin expression was observed(r =0.940,P ＜ 0.05).Conclusions These findings suggest that Wnt2/β-catenin signaling pathway may play important roles in the cardiovascular disease and be useful for exploring the molecular mechanism of myocardial injury..

Effects of Astragalus membranaous on the proliferation and transforming growth factorβ1 production of cardiac fibroblasts / 临床儿科杂志

ObjectiveTo observe the effect of Astragalus membranaous on angiotensinⅡ (AngⅡ)-induced transform-ing growth factor β1 (TGF-β1) production of cardiac ifbroblasts.Methods Cardiac ifbroblasts were culturedin vitro. Cells were allocated into 3 groups: control group, Astragalus membranaous groups (50, 100, 200 mg/ml), Ang II group (10-7 mol/L) and AngⅡ/Astragalus membranaous groups (50, 100, 200 mg/ml). The proliferation of each group was tested by methyl thiazolyl tetrazolium method. TGF-β1 was measured by ELISA.Results The proliferation of cardiac ifbroblasts had signiifcant difference between each groups (F=71.84,P=0.000). The proliferation of cardiac ifbroblasts with Ang II stimulation was higher than that of cells without Ang II stimulation (P<0.05). Astragalus membranaous inhibited Ang II-induced cardiac ifbroblasts proliferation dose dependently (P<0.05). The TGF-β1 production had signiifcant difference between each groups (F=786.81,P=0.000). The TGF-β1 production in AngII/astragalus membranaous groups was lower than that in Ang II group (P<0.05). The TGF-β1 production in Ang II group was the highest, and had signiifcant difference as compared to other groups (P<0.05). Astragalus membranaous inhibited Ang II-induced TGF-β1 production dose dependently (P<0.05).Conclusions Ang II can stimulate the proliferation of cardiac ifbroblasts, and promote the TGF-β1 production. Astragalus membranaous can inhibit the proliferation of Ang II-induced cardiac ifbroblasts, and reduce the TGF-β1 production of cardiac ifbroblasts.

From endoplasmic reticulum to Golgi apparatus: a secretory pathway controlled by signal molecules / 浙江大学学报·医学版

Protein transport from endoplasmic reticulum (ER) to Golgi apparatus has long been known to be a central process for protein quality control and sorting. Recent studies have revealed that a large number of signal molecules are involved in regulation of membrane trafficking through ER, ER-Golgi intermediate compartment and Golgi apparatus. These molecules can significantly change the transport rate of proteins by regulating vesicle budding and fusion. Protein transport from ER to Golgi apparatus is not only controlled by signal pathways triggered from outside the cell, it is also regulated by feedback signals from the transport pathway.

Study of Parkinson's disease based on Drosophila model / 浙江大学学报·医学版

Parkinson's disease (PD) is the second most common neurodegenerative disorder of the central nervous system, affecting 1 in 50 people over the age of 60 years. PD is the result of loss of a majority of dopamine (DA) neurons in the midbrain substantia nigra. The basic pathological feature of PD is the formation of intracytoplasmic eosinophilic inclusion bodies, Lewy bodies. So far, what leads to DA neuron degeneration is uncertain. Thirteen familial PD related loci have been identified, including six mutations:α-SYN(PARK1/4), Parkin(PARK2), UCHL1(PARK5), PINK1(PARK6), DJ-1(PARK7) and LRRK2 (PARK8). Drosophila has been widely used in the study of human diseases because of its genetic advantages. The Drosophila DA synthesis is similar to human, and Drosophila DA system is also involved in motion control, so it is generally considered that DA neuron death of Drosophila can be a perfect model of PD. In this article we review the progress of research methods based on Drosophila model in study of mechanisms related to Parkinson's disease.

Treatment and prognosis of patients with intrahepatic cholangiocarcinoma / 中国医师杂志

Objective To investigate the approaches for diagnosis and treatment of intrahepatic cholangiocarcinoma(ICC)and asgess its prognosis factors.Methods The clinical data of 86 patients with ICC in our hospital from January 1995 to December2005 were retrospeetively analyzed.All patients were divided into two groups according to the treatment method,including hepatectomy and lymphatic clearance group(Group A,n=42)and hepateetomy group(Group B,n=44),and their clinicopathological variables were analyzed.Resuits The 1-,3-and 5-year survival rates were 77.81%and 35.21%,20.93%and 19.82%,2.31%and 0%respectively between group A and group B.There was significantly difference between these two groups(P＜0.01).The analysis showed that resection and lymphatic clearance were correlated to prognosis.The 1-,3-and 5-year survival rates were 59.21%,26.21%,and 20.11% respectively in 47 patients who were found no lymph node metastasis,and the 1-,3-and 5-year survival rates were 19.82%,2.31%and 0% respectively in 39 patients who were found lymph node metastasis.There was significantly difference in survival rate between group A and group B(P＜0.01).Condusions Reseetability and lymphatic clearance are two significant factors correlated to survival of the patients with ICC.Aggresgive treatment of lymph node metastasis in hepatoduodenal ligament is an important strategy to improve survival rates and strengthen patient's life quality.

INFLUENCE OF PARTIALLY C-TERMINAL DELETION OF NR2A SUBUNITS ON THE SURFACE EXPRESSION AND FUNCTION OF NMDA RECEPTOR CHANNELS / 解剖学报

Objective To investigate whether the partially C-terminal deletion of NR2A subunit alters the surface expression and channel function of NMDA receptors in both HEK293 cells and cultured hippocampal neurons of rats. Methods Four plasmids for NR2A mutants with N-terminally GFP-tagged and C-terminal deletion NR2A?C1(?897L-1017S),NR2A?C2(?1024D-1142P),NR2A?C3(?1149D-1347G),and NR2A?C4(?1354S-1464V) were generated,and transfected into HEK293 cells and hippocampal neurons in culture.Surface staining was performed using anti-GFP antibody and Cy3 conjugated secondary antibody.Glutamate evoked currents were also detected using whole-cell recording. Results Positive surface staining was found for all the HEK293 cell co-transfected NR1-1a/NR2A?C1,NR1-1a/NR2A?C2,NR1-1a/NR2A?C3 or NR1-1a/NR2A?C4,and quantitative analysis showed no significant decrease in surface expression level when compared to that from NR1-1a/NR2A transfection.Glutamate-evoked currents were recorded in HEK293 cells co-transfected with NR1-1a/NR2A?C2 or NR1-1a/NR2A?C4.Surface expression level of NMDA receptor clusters on dendrites was significantly decreased in the neurons transfected with NR2A?C1,NR2A?C2 or NR2A?C3 than in those transfected with NR2A.Conclusion C-terminal deletion of NR2A subunit differentially effects surface expression of NMDA receptors in HEK293 cells and in hippocampal neurons in culture.

The synergistic effect of endothelin-1 and basic fibroblast growth factor on rat vascular smooth muscle cell proliferation / 中国病理生理杂志

AIM: The synergistic effect of basic fibroblast growth factor (bFGF) and endothelin-1 (ET-1) on rat aortic vascular smooth muscle cells(VSMC) proliferation was observed. The possible mechanism of the synergism was also investigated. METHODS: BrdU incorporation and cell counting method were adopted to value the pro-proliferative effect of VSMC. Western-blotting was used to observe the variation of bFGF and FGFR-1 isoforms expression. RESULTS: bFGF and ET-1 could promote VSMC proliferation separately, and synergistically in combination. The synergism was dose- and time-dependent. ET-1 increased all the three bFGF isoforms and FGFR-1 protein level in dose- and time-dependent manner. In addition, after exhaustion of intracellular PKC, the upregulation effects of ET-1 on bFGF and FGFR-1 expressions in VSMC both inclined. CONCLUSION: bFGF and ET-1 had synergistic effect on VSMC proliferation. ET-1 may increase the responsiveness of VSMC to bFGF through modulation of bFGF isoforms together with FGFR-1, which was PKC-dependent.

The synergistic effect of endothelin- 1 and basic fibroblast growth factor on rat vascular smooth muscle cell proliferation / 中国病理生理杂志

AIM: The synergistic effect of basic fibroblast growth factor (bFGF) and endothelin- 1 (ET-1 ) on rat aortic vascular smooth muscle cells(VSMC) proliferation was observed. The possible mechanism of the synergism was also investigated. METHODS: BrdU incorporation and cell counting method were adopted to value the pro - proliferative effect of VSMC. Western- blotting was used to observe the variation of bFGF and FGFR - 1 isoforms expression. RESULTS: bFGF and ET- 1 could promote VSMC proliferation separately, and synergistically in combination. The synergism was dose - and time - dependent. ET- 1 increased all the three bFGF isoforms and FGFR - 1 protein level in dose - and time- dependent manner. In addition, after exhaustion of intracellular PKC, the upregulation effects of ET- 1 on bFGF and FGFR - 1 expressions in VSMC both inclined. CONCLUSION: bFGF and ET- 1 had synergistic effect on VSMC proliferation. ET- 1 may increase the responsiveness of VSMC to bFGF through modulation of bFGF isoforms together with FGFR - 1, which was PKC - dependent.

Expression of GST fusion proteins of human cytochrome P 2B6 and preparation of anti-cytochrome P2B6 polyclonal antibody / 中国病理生理杂志

AIM: To get large amounts of pure antigens to raise specific antibodies and to perform quantifications.METHODS: CYP2B6 (cytochrome P) cDNA fragments was ligated into BamHI restricted PGEX-3b to generate recombinants PGEX/2B6. We identified recombinants PGEX/2B6 by EcoRI digestion. The expression of fusion proteins were induced by adding isopropyl-thiogalactoside(IPTG). Several clones showed high-level expression of fusion proteins. Insoluble proteins was isolated from the bacteria and the fusion proteins was recovered and purified from a preparative (2mm) SDS-PAGE. The polyarcrylamide gel containing the fusion proteins glutathione S-transferase(GST-2B6) were used to immunize BALB/C mice from which polyclonal ascites fluid was prepared. The purified fusion proteins GST-1A1(GST fusion protein of CYP1A1 cDNA246～386aa expressed in this library ,purified by preparative SDS-PAGE), GST-2B6 were used to test the specificity of 2B6pAb. RESULTS:Fusion proteins constructed between GST and CYP2B6 was expressed in Escherichia coli DH5?. Mouse antibodies are raised against the fusion proteins GST-2B6. 2B6pAb was fond to be specific antibody.CONCLUSION:Recombinant PGEX/2B6 were constructed and purified fusion proteins GST-2B6, and specific 2B6pAb were obtained.

Expression of phenylalanine ammonia-lyase cDNA from rice in E. coli BL21DE3 / 中国病理生理杂志

AIM: To study the expression and its kinetics of rice phenylalanine ammonia-lyase gene encoding into E. coli as the basis of treatment for phenylketouria. METHODS: The phenylalanine ammonia-lyase-1-cDNA(rPAL-1-cDNA) from rice was recombined into E. coli high expression vector pET-28c and transformed into E. coli host strain BL21DE3. Engineering bacteria was then inducted by isopropyl-?-D-thiogalactoside (IPTG) for 1, 3, 5, 7 hours, in order to obtain high level expression. RESULTS: After induction, the expression level of fusion protein was 21.40%, 30.60%, 35.40%, 35.43% respectively. The fusion protein exhibited a band of 78 6 kD on SDS-PAGE analysis,but was not found in controls.The target protein was mainly existed in the form of inclusion body. CONCLUSION:Rice PAL gene expressing E. coli was established by gentic engineering technique.

THE RESEARCH PROGRESS ON NG2 GLIAL CELLS / 解剖学报

Objective To review the physiological characters and potential function of chondroitin sulphate proteoglycan,CSPG(NG2) glial cells in the central nervous system. Methods In conclusion of the NG2 glial properties and the global, novel researches on NG2 cells. Results Cells that express the NG2 proteoglycan comprise a unique population of glial cells in the central nervous system.Study on their role in the CNS becomes more and more popular.Previous researches mainly focus on the differentiation and distinction.Now the relationship between NG2 glial cells and neurons,and the synaptic plasticity is becoming a hot topic.Conclusion This review referred the research development of NG2 glial cells,including the differentiation,migration,their interaction with neurons,and the potential role in glial cell regeneration in disease systematically.

SURFACE EXPRESSION AND LOCALIZATION OF NMDARECEPTOR ON DENDRITIC TREE OF HIPPOCAMPAL NEURONS IN CULTURE / 解剖学报

Objective To investigate the developmental profiles on surface expression and distribution of NMDA receptor clusters especially on dendritic structures in cultured hippocampal neurons of rats. Methods A vector expressing green fluorescent protein N-terminally tagged NR1a subunit(GFP-NR1a) was generated and transfected into cultured hippocamplal neurons at 5 days in vitro (DIV 5).Surface expressed GFP-NR1a containing NMDA receptor clusters was then labeled in living neurons by using anti-GFP primary antibody followed by Cy3-conjugated secondary antibody.Furthermore,distribution of the surface clusters was observed on the detailed dendritic structures visualized with co-expression of cyan fluorescent protein(CFP). Results Punctate green flourescence clusters can be seen throughout the neurons transfected with GFP-NR1a and surface staining of living neurons showed that the most of these flourescence clusters on dendritic arbors were surface expressed in mature neurons.In parallel,no significant difference was found in terms of density of the surface NMDA clusters on dendritic trees in neurons at different developmental stage.Interestingly,at DIV 7,surface NMDA receptor clusters were mostly observed on the dendritic shafts,but rarely on filopodia.In contrast,two weeks later about 50% of clusters were found located at the dendritic spines.Conclusion This study shows that surface NMDA receptor clusters are distributed in a differentiated way related to dendritic structures during neural development.Especially at very early developmental stage,no clusters was found presented on filopodia,instead they were extensively clustered with a density estimated to be not significantly different from mature neurons.Therefore,our results suggested that NMDA receptors are likely to be incorporated or recruited to postsynaptic sites in the form of pre-expressed surface cluster on dendritic shafts during glutamatergic synapse formation.

EFFECTS OF ?-FLUOROMETHYLHISTIDINE,AN IRREVERSIBLE INHIBITOR OF HISTIDINE DECARBOXYLASE,ON TURNING BEHAVIOR AND DOPANERGIC AND HISTAMINERGIC NEURONS IN 6-HYDROXYDOPAMINE-LESIONED RATS / 解剖学报

Objective To investigate the role of endogenous histamine in Parkinson disease. Methods 6-OHDA-lesioned rats were prepared by the conventional mothod,and in the meantime a group of rats were administrated with ?-fluoromethylhistidine(?-FMH),an irreversible inhibitor of histidine decarboxylase(HDC),via intracerebroventricular injection(12.5 ?g or 25 ?g,i.c.v.) for seven days.On the 7 day,the apomorphine-induced turning behavior and were detected, and the immunoreactivity of dopaminergic neurons in the substantia nigra pars compact(SNc) and histaminergic neurons in the tuberomammillary nucleus(TMN) were evaluated by tyrosine hydroxylase(TH) and HDC immunohistochemistry,respectively.Additionally,the level of dopamine in striatum was determined with high performance liquid chromatography(HPLC). Results ?-FMH(25 ?g,i.c.v.) significantly reduced the turning behavior and prevented the loss of dopaminergic neurons in the SNc,and slightly increased dopamine level in the striatum.Whereas,the immunoreactivity of histaminergic neurons in the TMN of hypothalamus in both the 6-OHDA lesioned and the ?-FMH treated rats was not changed.Conclusion Endogeneous histamine may involve in the pathological processes of PD.However,the histaminergic neurons are not involved in PD.

OPTIMIZATION OF THREE-CUBE FRET ASSAY AND APPLICATION IN THE ANALYSIS OF SUBUNIT ASSEMBLING OF iGluR RECEPTORS IN LIVING CELLS / 解剖学报

Objective To establish and optimize three-cube FRET assay in living cells and analyze subunit assembling of iGluR receptors. Methods Taking HEK293 cells cotransfed with pECFP and pEYFP as negative control, and those transfected with pECFP-YFP as positive control,different calculation methods using fluorescence microscopy were compared. Results These calculation methods were all suitable for FRET measurement in the system. but the measurement results were affected by the ratio of Donor/Acceptor (D/A) in some degree,and different calculation methods have different optimized conditions. FRET measurement using FR value showed subunit specific assembly of iGluR subtypes.Conclusion There are different optimized conditions for these different calculation methods in the three-cube FRET measurement system,and a further evidence is provided for subunit specific assembling of iGluR subtypes from the FRET assay.