Grik4 , Glutamate receptor, ionotropic, kainate 4; Grik5 , Glutamate receptor, ionotropic, kainate 5; Grin1 , Glutamate receptor, ionotropic, NMDA1; Grin3a , Glutamate receptor, ionotropic, NMDA3A; Grin3b , Glutamate receptor, ionotropic, NMDA3B; Grin2a , Glutamate receptor, ionotropic, NMDA2A; Grin2b , Glutamate receptor, ionotropic, NMDA2B; Grin2c , Glutamate receptor, ionotropic, NMDA2C; Grin2d , Glutamate receptor, ionotropic, NMDA2D; MW, molecular weight markers; (A) positive control tissue; (B) ovulated oocytes; (C) blastocysts. The MWs in base pairs (bp) are indicated to the right of the panels. * Primers for Grik3 , Grin2d , Grin3a, and Grin3b subunits were not included in the Mouse GABA Glutamate RT2 profiler PCR array, and commercial primer sets from Qiagen were used (see Materials and methods). Delta receptors (formed from delta 1 and delta 2 subunits), classified by sequence homology as ionotropic glutamate receptors, do not bind glutamate, and were not investigated in this study. " width="250" height="auto" />
Anti Grik4 Ka1 Extracellular Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Glutamate can act as a signaling molecule in mouse preimplantation embryos †"
Article Title: Glutamate can act as a signaling molecule in mouse preimplantation embryos †
Journal: Biology of Reproduction
Figure Legend Snippet: Transcripts encoding ionotropic glutamate receptors are expressed in mouse blastocysts and oocytes. Subunits of ionotropic glutamate receptors are listed in the table on the left. The most frequently used common names of subunits are given in parentheses. Mouse gene symbols are used (human gene symbols are the same but written with all letters capitalized, e.g., GRIA1 ). In transcripts which were consistently expressed in both blastocysts and oocytes, fold regulation values (“+" means upregulation and “-" means downregulation in blastocysts compared to oocytes) and corresponding P-values are shown. Transcripts were detected by reverse transcription-polymerase chain reaction (RT-PCR) and representative agarose gels with separated PCR products are shown in the panels on the right. Lanes: Gria1 , Glutamate receptor, ionotropic, AMPA1; Gria2 , Glutamate receptor, ionotropic, AMPA2; Gria3 , Glutamate receptor, ionotropic, AMPA3; Gria4 , Glutamate receptor, ionotropic, AMPA4; Grik1 , Glutamate receptor, ionotropic, kainate 1; Grik2 , Glutamate receptor, ionotropic, kainate 2; Grik3 , Glutamate receptor, ionotropic, kainate 3; Grik4 , Glutamate receptor, ionotropic, kainate 4; Grik5 , Glutamate receptor, ionotropic, kainate 5; Grin1 , Glutamate receptor, ionotropic, NMDA1; Grin3a , Glutamate receptor, ionotropic, NMDA3A; Grin3b , Glutamate receptor, ionotropic, NMDA3B; Grin2a , Glutamate receptor, ionotropic, NMDA2A; Grin2b , Glutamate receptor, ionotropic, NMDA2B; Grin2c , Glutamate receptor, ionotropic, NMDA2C; Grin2d , Glutamate receptor, ionotropic, NMDA2D; MW, molecular weight markers; (A) positive control tissue; (B) ovulated oocytes; (C) blastocysts. The MWs in base pairs (bp) are indicated to the right of the panels. * Primers for Grik3 , Grin2d , Grin3a, and Grin3b subunits were not included in the Mouse GABA Glutamate RT2 profiler PCR array, and commercial primer sets from Qiagen were used (see Materials and methods). Delta receptors (formed from delta 1 and delta 2 subunits), classified by sequence homology as ionotropic glutamate receptors, do not bind glutamate, and were not investigated in this study.
Techniques Used: Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Molecular Weight, Positive Control, Sequencing
2) Product Images from "Thalamic circuits for independent control of prefrontal signal and noise"
Article Title: Thalamic circuits for independent control of prefrontal signal and noise
Figure Legend Snippet: Supportive evidence for anatomical and functional segregation of the two MD cell types. a-c , Left: Starter neurons (arrowheads) in the PL of VIP-cre ( a ), PV-cre ( b ) and SST-cre ( c ) mice for monosynaptic retrograde tracing using rabies viruses. Right: Starter neurons identified by co-expression of TVA fused to GFP (top) and blue fluorescent protein from rabies viruses (bottom) in VIP, PV, and SST neurons, respectively. Scale bars in µm: 200 µm(left), 30 µm (right). d , Left: Representative images of MD neurons that monosynaptically target PL SST+ interneurons. Scale Bar in µm: 200. (Note a lack of preferential localization within the MDl) Right: 3D plot of the anatomical location of SST-projecting MDl neurons (n = 86 SST projecting neurons from 3 mice). e , Anatomical separation between SST-projecting MD neurons and VIP-/PV- projecting MD neurons quantified as high misclassification using KNN clustering. f , MD D2 and MD GRIK4 neuronal locations show low misclassification compared to VIP- and PV-projecting MD neurons respectively. g , Example of a PL neuron showing amplification of evoked responses through concurrent intra-PL and MD D2 optical stimulation (left), but not when intra-PL stimulation is combined with MD GRIK4 stimulation (right). h , Examples of excitatory (RS) and inhibitory (FS) PL neurons showing, respectively, suppression and increase in spike rates with optical activation of MD GRIK4 neurons but not with activation of MD D2 neurons. i , Parametric activation of MD GRIK4 , but not MD D2 , neurons increase spike rates of PL inhibitory neurons (n = 68 and n = 78 PL-FS neurons from 3 animals each of MD D2 and MD GRIK4 respectively; p = 0.874, For MD GRIK4 p = 0.556, *p = 0.0387, ***p = 9.36 x 10 -6 , *p = 0.0387 respectively for laser powers 0.65, 1.3, 3.5 and 7.0 mW/mm 2 ; Mann-Whitney U test compared to baseline). j , left: D2 specific promoter (D2SP) driven expression of mCherry + (CreON) and co-expression of EYFP (CreOFF) in Cre-negative neurons using a Cre - Out intersectional strategy labels two populations similar to D2-cre and Grik4-cre, but in WT animal. Right: Magnified images showing mCherry (D2SP+) and eYFP (Cre negative) neurons. Scale bar = 200 µm k , Consistent anatomical similarity between MD D2SP and MD D2 populations and a corresponding segregation between MD D2SP and MD GRIK4 neurons, quantified using representational similarity analysis (n = 95 cells from 2 animals for MD D2SP ). l , A comparable similarity and segregation as shown in ( k ) is found when comparing MD D2SP neurons to VIP-projecting and PV projecting neurons. m , top row: MD D2 Cre-expressing neurons (MD D2+ ) labelled with GFP have extremely sparse Grik4 protein expression (IHC) compared to MD D2 Cre-negative (MD D2 - neurons (middle row) or MD GRIK4 expressing neurons (bottom row). Scale bar = 3 µm n , Quantification of data (n = 116 MD D2+ , 106 MD D2- and 124 MD GRIK4 neurons from 2 animals, demonstrating substantial Grik4 immunolabelling overlap between D2- and Grik4+ neural populations (not significantly different), but both being different from the D2+ population. ***p = 0.0001 for both comparisons, Kruskal Wallis test). o , Direct comparison of Grik4 immunolabelling across D2+ and D2- neurons (thresholded by the lowest 10th percentile of this analysis puts an upper bound estimate of 15% overlap between the D2+ and Grik4+ population. ‘positive control’ Grik4+ neurons). All statistical tests are two-tailed. For box plot n boundaries, 25–75th percentiles; midline, median; whiskers, minimum–maximum. Data are presented as mean ± SEM for i Source data .
Techniques Used: Functional Assay, Mouse Assay, Retrograde Tracing, Expressing, Amplification, Activation Assay, MANN-WHITNEY, Immunohistochemistry, Positive Control, Two Tailed Test
Figure Legend Snippet: Differential engagement of the two MD cell types in a PL-dependent behaviour. a , Schematic illustration of PL dependent attention control task (see Methods ). In brief, on each trial, animals have to remember a 100ms HP or a LP auditory cue over a delay period to execute the corresponding rule (HP - attend to audition vs LP – attend to vision) and make a choice to either follow a target stimulus (auditory vs visual) to collect a milk reward. Orange bars highlight the two epochs where MD is optically inhibited across the experiment types b , Illustration of PL independent task with a 2AFC design, where animals have to respond to the side a LED light target was presented (without a distractor). c , Example sessions plotting the performance of an animal in the PL dependent (black) and PL independent (grey) versions of the task. d , Well trained animals show a daily ramp-up of performance in the first 30 trials of the PL dependent (but not PL independent) task, starting from chance (trials 1 to 10) and progressing onto performance > 0.7 proportion correct (‘task engagement’, trials 20 to 30) (n = 12 sessions over 4 mice, ***p = 0.0002,). e , f , optical MD inactivation (yellow) during the cueing period of the first 30 trials in a session prolongs the ramp up to task engagement in the PL dependent task ( e ) but not the PL independent task (n = 12 sessions over 4 mice, p = 0.0008, Kolmogorov-Smirnov test). g , Optical inactivation of MD D2 neurons have no effect on number of trials taken to task engagement in PL dependent attention control task (n = 12 sessions over 4 D2-cre mice, p = 0.466 (NS), comparing across laser ON vs laser OFF sessions). h , Optical inactivation of MD GRIK4 neurons increase the number of trials required to reach task engagement in the attention control task (n = 12 sessions over 4 GRIK4-cre mice, ***p = 2.2 x 10 -6 , comparing across laser ON vs laser OFF sessions). i , Optical MD inactivation during the cueing period of the first 30 trials delays task engagement (performance at > 0.7 proportion correct; n = 12 sessions over 4 animals, ***p= 1.5 x 10 -6 ;). j-k , Optical MD GRIK4 (but not MD D2 ) inactivation recapitulates the effect in b (n = 12 sessions each from 4 D2-cre and 4 GRIK4-cre mice; ***p = 2.9 x 10 -5 , p = 0.7657 (NS);). l , Optical MD inactivation in the delay period of
Techniques Used: Mouse Assay
Figure Legend Snippet: Two MD circuits for amplification and suppression of PFC activity. a , Prefrontal PV + and VIP + input mapping. b , MD neurons targeting VIP + (left) and PV + (right) interneurons occupy distinct MDl domains. Scale bars, 200 µm. c , Group summary for location of VIP- and PV-projecting MDl neurons ( n = 73 VIP-projecting (7 mice) and n = 117 PV-projecting (4 mice)). d , KNN clustering and representational similarity analysis show robust separation. e , Labelling MD D2 and MD GRIK4 neurons using the corresponding Cre lines. f , MD D2 and MD GRIK4 neurons also occupy distinct anatomical locations. Scale bars, 200 μm. g , Group summary of MD D2 and MD GRIK4 neurons in MDl ( n = 177 MD D2 neurons and 194 MD GRIK4 neurons from 3 mice each). h , MD D2 and MD GRIK4 locations show additional high representational similarity to VIP- and PV-projecting neurons, respectively. i , j , mGRASP labelling shows higher innervation of VIP + neurons by MD D2 ( i ; n = 21 neurons from 3 D2-cre, n = 25 neurons from 3 GRIK4-cre mice, respectively; Kolmogorov–Smirnov) and higher innervation of PV + neurons by MD GRIK4 ( j ; n = 27 neurons from 3 D2-cre, n = 32 neurons from 3 GRIK4-cre mice, respectively; Kolmogorov–Smirnov). Scale bars, 3 µm. k , Hypothesized circuit. l , Selective MD cell-type activation set-up. m , MD D2 but not MD GRIK4 amplify functional PL connectivity ( n = 100 and n = 68 PL neural responses from 3 mice each for MD D2 and MD GRIK4 , respectively; left to right: MD D2 P = 1.0 × 10 −5 for all; MD GRIK4 P = 0.0599, 0.0789, 0.0575, 0.1311 (NS) for laser powers displayed; Mann-Whitney U , compared to baseline). n , MD GRIK4 but not MD D2 suppress PL neural spike rates ( n = 1,257 and n = 697 putative excitatory PL neurons from 3 mice each; MD D 2 P = 0.184, 0.605, 0.579, 0.739 (NS); MD GRIK4 P = 0.298, P = 0.067, * P = 0.033, *** P = 1.61 × 10 −5 , respectively, for laser powers displayed; Mann-Whitney U compared to baseline). All statistical tests are two-tailed. Box plot parameters as in Fig. 1. Data are mean ± s.e.m. for m , n . Source data .
Techniques Used: Amplification, Activity Assay, Mouse Assay, Activation Assay, Functional Assay, MANN-WHITNEY, Two Tailed Test
Figure Legend Snippet: Untagged neurons in the tagging experiments are no different than generic recordings, and optical inhibition of terminals of the two cell types replicates cell body inactivation. a , (Top Left) Schematic of optogenetic tagging and identification of MD D2 and MD GRIK4 neurons. MD D2 or MD GRIK4 neurons are tagged with NpHR3.0 and identified via light activated spike rate suppression. (Bottom) Example tagged neuronal response to NpHR3.0 activation. (Right) Tagged neurons from one mouse (red) are identified using k-means clustering (features: change in firing rate, proportion of trials suppressed, and half-time to recover from suppression (n = 262 total number of neurons). b , Relative fraction of all MD neurons from GRIK4-cre mice that are conflict-preferring vs. non-preferring are comparable to that of wild-type animals (Fig. 3g ) (n = 91 neurons from 3 mice; p = 0.429 (NS), chi-squared test). Note that tagged MD GRIK4 neurons are significantly more conflict-preferring compared to the whole population (Fig. 4b ) (p = 0.0175; chi-squared test). c , Relative fraction of all MD neurons from D2-cre mice that are conflict-preferring vs. non-preferring, are also comparable to that of wild-type animals (Fig. 3g ) (n = 95 neurons from 3 mice; p = 0.166 (NS), chi-squared test). Note, that tagged MD D2 neurons are significantly more conflict-non-preferring (Fig. 4d ) (p=1.34 x 10 -4 ; chi-squared test). d , Optical inhibition of MD GRIK4 terminals in the PL recapitulates the loss in task accuracy across low and high conflict trials as seen with optical MD GRIK4 inactivation (Fig. 4e ; n = 20 sessions over 4 GRIK4-cre mice, *p = 0.0199, ***p = 0.0002; Mann-Whitney U test). e , Optical inhibition of MD D2 terminals in the PL enhances performance accuracy on trials with high cueing conflict, similar to the effect of optical MD D2 inactivation (Fig. 4f ; n = 20 sessions over 4 D2-cre mice, p = 0.3941 (NS), **p = 0.0023; Mann-Whitney U test. f , Schematic of micro-drive bottom piece and the 3x3 grid organization of the tetrode array for MD recordings. g , Summary of the density of tagged neurons on the medial-lateral axis separated by animal. We show the result for 2 Grik4-cre (top and bottom) and 2 D2-cre animals ((top and bottom rows) that have enough numbers of tagged neurons. All statistical tests are two-tailed. For box plots b – e boundaries, 25–75th percentiles; midline, median; whiskers, minimum–maximum Source data .
Techniques Used: Inhibition, Activation Assay, Mouse Assay, MANN-WHITNEY, Two Tailed Test
Figure Legend Snippet: The two thalamic cell types are engaged by different task input statistics. a , Example tagged MD GRIK4 neuron recorded in the task. b , Tagged MD GRIK4 neurons are more likely to be conflict-preferring ( n = 17 neurons from 3 mice; P = 0.0042; binomial). c , Example tagged MD D2 neuron recorded in the task. d , Tagged MD D2 neurons are more likely to be conflict-non-preferring ( n = 20 neurons from 3 mice; P = 4.0 × 10 −5 ; binomial). e , MD GRIK4 suppression recapitulates generic MD suppression ( n = 20 sessions from 4 GRIK4-cre mice; Wilcoxon signed-rank). f , MD D2 inactivation enhances performance accuracy on trials with high cueing conflict ( n = 20 sessions from 4 D2-cre mice; Wilcoxon signed-rank). g , Expanded neural model with two MD cell types. h , The two-cell-type model captures experimental data ( n = 2,000 trials, chi-squared). MD+, MD intact; G−, without GRIK4; D−, without D2. i , Stimulus configuration for sparseness-driven uncertainty. j , Performance accuracy is modulated by cueing sparseness, and optical MD deafferentiation diminishes performance on trials with higher cueing sparseness ( n = 25 sessions, 4 mice; * P = 0.0222, *** P = 1.02 × 10 −4 ; chi-squared). k , MD GRIK4 inactivation improves performance accuracy on both high and low signal trials ( n = 20 sessions from 4 GRIK4-cre mice; Wilcoxon signed-rank). l , Optical MD D2 inactivation recapitulates optical generic MD deafferentiation ( n = 20 sessions from 4 D2-cre mice; Wilcoxon signed-rank). All statistical tests are two-tailed. Box plot parameters as in Fig. 1 . Data are mean ± s.e.m. for h , j . Source data .
Techniques Used: Mouse Assay, Two Tailed Test
Figure Legend Snippet: Basic and extended mean-field models. a , Schematic of the mean-field neural model that describes generic MD inactivation results (see Extended Data Fig. 7o ). The model describes two PL populations that receive separate inputs corresponding to the cues in favour of the two attentional rules (HP - attend to vision or LP – attend to audition). Each population has strong recurrent self-excitation and net inhibition on the other population. The MD component of the model receives inputs from the PL (see Extended Data Fig. 7 ) and is activated by conflict to inhibit the two PL populations. b , Example model decision variables in a trial early biased to the wrong attentional choice, demonstrating how MD-mediated suppression may improve performance of the model. When MD is intact (left), strong early evidence to the wrong choice (high-pass in this example; cueing sequence in inset) increases the decision variable of the non-preferred population early on, but the preferred population prevails when the preferred stimulus dominates in the latter half of the cueing sequence. On the other hand, in the absence of MD conflict-driven suppression of cue integration in the PL (right), the early non-preferred inputs drive the non-preferred population to maintain high activity, suppressing the preferred population’s response to late inputs. c , Schematic of the mean-field neural model incorporating the two cell types, where MD GRIK4 is conflict-activated and suppresses PL, and MD D2 is conflict-suppressed and amplifies PL recurrence. MD D2 results in enhanced gain of the PL input-output function (bottom). d , Example model decision variables for high conflict trials, with (left) and without (right) MD D2 . Increased PL recurrence due to MD D2 results in larger response to input cues. However, the effect is less pronounced for preferred cues as the population activity and decision variable saturate with inputs. As a result, the larger response to input cues asymmetrically favours the non-preferred population, and the separation between preferred and non-preferred activity is larger without MD D2 (shown are median over 1,000 trials). e , Example model decision variables for low signal sparse trials (Fig. 4 ), with (left) and without (right) MD D2 module. Increased PL recurrence due to MD D2 allows amplified response of the preferred population to sparse input cues, but minimally affects the non-preferred population which receives no input cues. As such, MD D2 results in a larger separation between preferred and non-preferred activity (shown are median over 1,000 trials).
Techniques Used: Inhibition, Sequencing, Activity Assay, Amplification
Figure Legend Snippet: mGRASP and synaptophysin labelling provide evidence for output segregation of the two MD cell types. a , Cartoon depicting strategy to label cell type specific MD→PL thalamocortical synapses using mGRASP. The pre mGRASP component is virally expressed in MD D2 or MD GRIK4 neurons in the respective Cre lines while the post mGRASP component is ubiquitously expressed in the PL. MD D2 or MD GRIK4 specific mGRASP synapses onto VIP vs PV neurons in the PL are identified by immunohistochemistry guided detection of PV and VIP neurons expressing post mGRASP in the PL. b–c , Left: Representative images of MD D2 ( b ) and MD GRIK4 ( c ) neurons expressing pre mGRASP in the MD of D2-cre and GRIK4-cre mice respectively. Right: Ubiquitous expression of post mGRASP+ neurons detected by TdTomato fluorescence in the PL of D2-cre ( b ) and GRIK4-cre ( c ) mice. Scale bar in µm: 200. d , Left to right: Examples of PL VIP+ neurons showing post mGRASP expression (magenta), VIP expression detected via immunohistochemistry (yellow) and mGRASP+ synapses from MD D2 (cyan dots, top row) or MD GRIK4 (cyan, dots, bottom row) neurons. e , Same as in d , for PL PV+ neurons. Scale bars in µm: 3 µm. f , Representative images showing layer-wise termination of synapses from MD D2 (left) and MD GRIK4 (right) neurons in the PL, labelled with virally expressed GFP fused to synaptic protein (synaptophysin). Scale bar in µm: 100. g , MD D2 neurons terminate in L1 of the PL with a higher frequency compared to MD GRIK4 neurons (n = 12 sections each from 3 D2-cre and 3 GRIK4-cre mice, *p = 0.0253, *p = 0.039, two-tailed Mann-Whitney U test comparing 50 µm bins from the pial surface across groups). All statistical tests are two-tailed. Data are presented as mean ± SEM for g Source data .
Techniques Used: Immunohistochemistry, Expressing, Mouse Assay, Fluorescence, Two Tailed Test, MANN-WHITNEY