Journal: eLife

Article Title: Reliability of an interneuron response depends on an integrated sensory state

doi: 10.7554/eLife.50566

Figure Lengend Snippet: ( A and B ) AWA GCaMP2.2b ( A ) or AIA GCaMP5A ( B ) calcium responses to 10 s pulses of increasing concentrations of diacetyl and to AWA optogenetic stimulation. Bold lines indicate mean response, and light lines show individual traces. AWA traces to optogenetic stimulation were randomly downsampled to 40 traces from a complete set of 268 traces to match the number of odor traces and enhance visibility. AIA traces were randomly downsampled to 10 traces from a set of 34 (for 0–1.15 µM diacetyl) or 569 (for AWA::Chrimson stimulation). In all schematic diagrams, calcium was monitored in the neuron indicated in green, resistor symbols represent gap junctions, and thin arrows represent chemical synapses. ( C and D ) Heat maps of AWA ( C ) or AIA ( D ) calcium traces from ( A ) and ( B ), respectively. Responses to optogenetic stimulation were downsampled to 32 traces ( C ) or 34 traces ( D ) for visibility and to match sample sizes to diacetyl; see for complete data. Each heat map row represents a calcium trace to a single stimulus pulse; each animal received two stimulus pulses. Traces are ordered by response latency. ( E ) Representative AIA calcium traces to a given stimulus. Responses were sorted by response latency, binned into ten bins, then one trace was randomly selected from each bin for presentation. ( F ) Cumulative response time profiles of AWA and AIA responses representing response latencies and probability, without downsampling. Only first 5 s of stimulation are shown. Arrows indicate the delay between the time at which 50% of AWA neurons responded versus the time at which 50% of AIA neurons responded. Figure 1—source data 1. Source data for and figure supplements.

Article Snippet: We used Metamorph 7.7.6 software to control image acquisition and light pulsing in addition to rapid stimulus switching (National Instruments NI-DAQmx connected to an Automate Valvebank 8 II actuator that controls a solenoid valve), odor selection (Hamilton 8-way distribution valve), and activation of an external red LED for Chrimson stimulation (Mightex Precision LED Spot Light, 617 nm, PLS-0617–030 s; attached to Chroma ET605/50x filter to narrow band to 605 ± 25 nm).

Techniques:

Journal: eLife

Article Title: Reliability of an interneuron response depends on an integrated sensory state

doi: 10.7554/eLife.50566

Figure Lengend Snippet: ( A ) Schematic of experimental configuration. Animals are paralyzed in a microfluidic device and their neural activity is recorded during exposure to odor or light. Two arenas can be recorded simultaneously with up to 10 animals per arena. See Materials and methods for details. ( B and C ) AWA requires both retinal pre-treatment and expression of the Chrimson transgene for responses to 617 nm light. ( B ) Mean AWA calcium responses; shading indicates ± SEM. Transgene with retinal: n = 74; transgene without retinal: n = 48; retinal without transgene: n = 16. ( C ) Cumulative response time profiles of data from ( B ), showing first 5 s of light exposure. ( D ) Individual AWA calcium responses to direct Chrimson excitation (617 nm) using various 474 nm light levels to excite GCaMP. Under strong illumination of GCaMP with 474 nm light (full power of LED, 165 mW/cm 2 ), GCaMP fluorescence did not increase with subsequent 617 nm stimulation, presumably because of direct Chrimson excitation by 474 nm light. At lower 474 nm light levels (15–65 mW/cm 2 ), or using a duty cycle with 10 ms of illumination every 100 s (10% of 40–165 mW/cm 2 ), GCaMP fluorescence transiently increased but returned to a baseline that allowed a subsequent Chrimson response to 617 nm light. Chrimson responses were partly suppressed under continuous 474 nm illumination or at the highest light level at 10% illumination, suggesting some cross-activation. The 10% duty cycle at 40 mW/cm 2 minimized the initial transient response to 474 nm while providing a strong signal to noise ratio at 617 nm. The 10% duty cycle at 15 mW/cm 2 was too dim for AIA GCaMP experiments. In general, the 10 ms/100 ms duty cycle for 474 nm illumination is used for GCaMP experiments because strobing reduces motion artefacts, and because the duty cycle minimizes GCaMP photobleaching during long-term imaging. AWA had normal responses to 1.15 µM diacetyl when using the 10% duty cycle of 40 mW/cm 2 ; this light level was used for all Chrimson experiments. Left: n = 20–25; Right: AWA::Chr, with retinal: n = 45; No Chrimson, no retinal: n = 16. ( E ) AWA calcium responses to four consecutive pulses of Chrimson excitation. AWA either responded to all or none of the Chrimson pulses, suggesting that variability in AWA responses to Chrimson is the property of an individual animal rather than a trial-to-trial property. Note that all experiments in other figures used two stimulus pulses, unless otherwise stated. ( F ) AWA::Chrimson::sl2::mCherry transgene expression levels for four separate experiments. Animals that responded to Chrimson excitation are in gray; animals that did not respond are in orange. There is no obvious correlation between mCherry expression level and the likelihood of AWA responses to light.

Article Snippet: We used Metamorph 7.7.6 software to control image acquisition and light pulsing in addition to rapid stimulus switching (National Instruments NI-DAQmx connected to an Automate Valvebank 8 II actuator that controls a solenoid valve), odor selection (Hamilton 8-way distribution valve), and activation of an external red LED for Chrimson stimulation (Mightex Precision LED Spot Light, 617 nm, PLS-0617–030 s; attached to Chroma ET605/50x filter to narrow band to 605 ± 25 nm).

Techniques: Activity Assay, Expressing, Fluorescence, Activation Assay, Imaging

Journal: eLife

Article Title: Reliability of an interneuron response depends on an integrated sensory state

doi: 10.7554/eLife.50566

Figure Lengend Snippet: ( A ) Cumulative response time profiles of AIA responses to 11.5 nM diacetyl in WT versus unc-13(e51) and unc-18(e234) animals (synaptic transmission mutants). ( B ) Heat maps of AIA responses to AWA::Chrimson stimulation in WT and unc-18(e234) animals. WT data were randomly downsampled to 57 traces for visibility and to match number of unc-18(e234) traces; data shown for single experiment block. See for pooled data from all experiments. ( C ) Cumulative response time profiles of AIA responses to AWA::Chrimson stimulation in WT versus unc-13, unc-18(e234), and unc-18(e81) animals. ( D ) Cumulative response time profiles of WT and unc-18(e234) response time profiles, combined over all experiments. Thick lines represent distributions of all data, faint lines represent distributions from individual experimental blocks. ( E ) Cumulative response time profiles of AWA responses to AWA::Chrimson stimulation in WT versus unc-13 animals. ( F ) Cumulative response time profiles of AWA and AIA responses to AWA::Chrimson stimulation in unc-13 animals. Arrow indicates the delay between the time at which 50% of AWA versus 50% of AIA neurons have responded. ( G ) Delay between the time at which 50% of AWA versus 50% of AIA neurons responded to 1.15 µM diacetyl and AWA::Chrimson stimulation in WT and unc-13 animals. WT responses are the same as in . Bars are mean ± SEM. Asterisks refer to Kolmogorov-Smirnov test significance versus WT over full 10 s stimulus pulse. ns: not significant; *: p<0.05; ***: p<0.001. See for sample sizes and test details. Additional heat maps of data from appear in . Figure 3—source data 1. Source data for and figure supplement.

Article Snippet: We used Metamorph 7.7.6 software to control image acquisition and light pulsing in addition to rapid stimulus switching (National Instruments NI-DAQmx connected to an Automate Valvebank 8 II actuator that controls a solenoid valve), odor selection (Hamilton 8-way distribution valve), and activation of an external red LED for Chrimson stimulation (Mightex Precision LED Spot Light, 617 nm, PLS-0617–030 s; attached to Chroma ET605/50x filter to narrow band to 605 ± 25 nm).

Techniques: Transmission Assay, Blocking Assay

Journal: eLife

Article Title: Reliability of an interneuron response depends on an integrated sensory state

doi: 10.7554/eLife.50566

Figure Lengend Snippet: ( A – C ) Magnitude of individual AWC ( A ), AWA ( B ), and ASK ( C ) responses to buffer (0), 0.9 µM, 9 µM, and 9 µM isoamyl alcohol shown in . Boxes show median and interquartile range. ( D – F ) Cumulative response time profiles of responses from ( A ), ( B ), and ( C ). Only the first 5 s are shown. ( G – J ) Cumulative response time profiles of AWC, AWA, AIA, and ASK (for J only) responses to 0.9 µM ( G ), 9 µM ( H ), or 90 µM ( I ) isoamyl alcohol, or 1.15 µM diacetyl ( J ) in WT animals. ( K ) Heat maps of AIA responses to 0.9, 9, or 90 µM isoamyl alcohol in WT versus odr-7 animals (AWA cell fate mutants), ceh-36 animals (AWC and ASE cell fate mutants), and odr-7 ceh-36 animals, shown in . ( L ) Mean AIA responses to 1.15 µM diacetyl, 90 µM isoamyl alcohol, AWA::Chrimson, and E. coli OP50 bacteria-conditioned medium from that initiated within 5 s of stimulus, aligned to the beginning of AIA activation rather than stimulus time. Shading indicates ± SEM. 1.15 µM diacetyl: n = 390; 90 µM isoamyl alcohol: n = 76; AWA::Chrimson: n = 260; OP50: n = 37. ( M ) Rise time of responses from ( M ). Bars indicate ± SEM. For ( A ), ( B ), ( C ), ( L ), and ( M ) asterisks refer to statistical significance of a one-way ANOVA with Dunnett’s multiple comparisons test versus buffer magnitude ( A–C ), versus 1.15 µM diacetyl magnitude ( L ), or versus 1.15 µM diacetyl rise time ( M ). ns: not significant; *: p<0.05; ***: p<0.001. See ( A–C and L ) or ( M ) for sample sizes and test details. For ( D–J ), asterisks refer to statistical significance of a Kolmogorov-Smirnov test versus buffer ( D–F ), between AWA and AIA ( G–I ), or between ASK and AIA ( J ), over full 10 s stimulus pulse. ns: not significant; *: p<0.05; **: p<0.01; ***: p<0.001. See for sample sizes and test details.

Article Snippet: We used Metamorph 7.7.6 software to control image acquisition and light pulsing in addition to rapid stimulus switching (National Instruments NI-DAQmx connected to an Automate Valvebank 8 II actuator that controls a solenoid valve), odor selection (Hamilton 8-way distribution valve), and activation of an external red LED for Chrimson stimulation (Mightex Precision LED Spot Light, 617 nm, PLS-0617–030 s; attached to Chroma ET605/50x filter to narrow band to 605 ± 25 nm).

Techniques: Bacteria, Activation Assay

Journal: eLife

Article Title: Reliability of an interneuron response depends on an integrated sensory state

doi: 10.7554/eLife.50566

Figure Lengend Snippet: ( A ) Heat maps of AWA and AIA responses to AWA::Chrimson stimulation, combined over all experiments. AWA: n = 268; AIA: n = 569. ( B ) Delay between the time at which 50% of AWA versus 50% of AIA neurons responded to various stimuli. Delay was greatest for AWA::Chrimson stimulation. Bars are mean ± SEM. ( C ) Cumulative response time profiles of AIA responses to 1.15 µM diacetyl recorded immediately after recordings of AWA::Chrimson stimulation (blue), representing a subset of animals used in ( A ). AIA responses to 1.15 µM diacetyl combined over all experiments are shown for comparison (black). ( D ) Response latencies of 318 AIA responses to AWA::Chrimson stimulation do not correlate with GCaMP fluorescence levels at pre-stimulus baseline. ( E ) Response latencies of 31 responses to AWA::Chrimson stimulation do not correlate with Chrimson transgene expression levels. ( F ) Representative AIA calcium traces to two pulses of AWA::Chrimson stimulation. AIA can response to the first pulse only, second pulse only, both pulses, or neither pulse. ( G, K, and O ) Cumulative response time profiles of AIA responses to the first or second stimulation with AWA::Chrimson ( I ), 11.5 nM diacetyl ( L ), or 1.15 µM diacetyl ( O ). Only animals for which both trial pulses yielded usable results were included (e.g. for AIA::Chrimson, 282/287 animals). All other figures and analyses beyond this supplement pool responses to both pulses. ( H, L, and P ) Proportion of animals that respond to only the first, second, both, or neither pulse of stimulation with AWA::Chrimson ( H ), 11.5 nM diacetyl ( L ), or 1.15 µM diacetyl ( P ). Some of the 22% of animals that did not respond to either AWA::Chrimson pulse may be the result of AWA::Chrimson transgene failure (~15% failure rate; see ); transgene failure does not explain the large proportion of animals that responded to one of two pulses, nor does it explain variability in response latencies. ( I and J ) In 98 animals that responded to both AWA::Chrimson stimulation pulses, there was no correlation between response latencies across pulses ( I ), but response magnitudes were correlated across pulses ( J ). ( M and N ) In 72 animals that responded to both 11.5 µM diacetyl pulses, there was no correlation between response latencies across pulses ( M ), but response magnitudes were correlated across pulses ( N ). ( Q and R ) In 187 animals that responded to both 1.15 µM diacetyl pulses, there was a moderate correlation between response latencies ( Q ) and a correlation between response magnitudes ( R ) across pulses. Three outlier data points were excluded from ( R ) but were included in analysis. For ( C ), ( G ), ( K ), and ( O ), asterisks refer to Kolmogorov-Smirnov test significance over full 10 s stimulus pulse. ns: not significant; *: p<0.05; **: p<0.01; ***: p<0.001. See for sample sizes and test details. For ( D ), ( E ), ( I ), ( J ), ( M ), ( N ), ( Q ) and ( R ), asterisks refer to significance of linear regression slope differing from 0. ns: not significant; ***: p<0.001.

Article Snippet: We used Metamorph 7.7.6 software to control image acquisition and light pulsing in addition to rapid stimulus switching (National Instruments NI-DAQmx connected to an Automate Valvebank 8 II actuator that controls a solenoid valve), odor selection (Hamilton 8-way distribution valve), and activation of an external red LED for Chrimson stimulation (Mightex Precision LED Spot Light, 617 nm, PLS-0617–030 s; attached to Chroma ET605/50x filter to narrow band to 605 ± 25 nm).

Techniques: Comparison, Fluorescence, Expressing

Journal: eLife

Article Title: Reliability of an interneuron response depends on an integrated sensory state

doi: 10.7554/eLife.50566

Figure Lengend Snippet: ( A ) Cumulative response time profiles of AIA to 1.15 µM diacetyl in WT versus unc-13(e51) , unc-18(e234) , and unc-18(e81) animals (synaptic transmission mutants). ( B ) Heat maps of AIA responses to AWA::Chrimson stimulation in WT and unc-18(e234) animals, combined over all experiments. WT: n = 569; unc-18(e234) : n = 335. ( C ) Heat maps of AWA responses to AWA::Chrimson stimulation in WT and unc-13 animals from . ( D ) Magnitude of AWA responses shown in ( C ), omitting traces that did not produce a detectable response. Boxes show median and interquartile range. ( E ) Cumulative response time profiles of AIA responses to AWA::Chrimson stimulation in WT versus unc-31 animals (dense core vesicle exocytosis mutants). For ( A ) and ( E ), asterisks refer to Kolmogorov-Smirnov test significance versus WT over full 10 s stimulus pulse. ns: not significant; **: p<0.01. See for sample sizes and test details. For ( D ), asterisks refer to statistical significance of an unpaired t-test. ns: not significant; **: p<0.01. See for sample sizes and test.

Article Snippet: We used Metamorph 7.7.6 software to control image acquisition and light pulsing in addition to rapid stimulus switching (National Instruments NI-DAQmx connected to an Automate Valvebank 8 II actuator that controls a solenoid valve), odor selection (Hamilton 8-way distribution valve), and activation of an external red LED for Chrimson stimulation (Mightex Precision LED Spot Light, 617 nm, PLS-0617–030 s; attached to Chroma ET605/50x filter to narrow band to 605 ± 25 nm).

Techniques: Transmission Assay

Journal: eLife

Article Title: Reliability of an interneuron response depends on an integrated sensory state

doi: 10.7554/eLife.50566

Figure Lengend Snippet: ( A ) Proportion of AIA calcium traces counted as ‘responses’ at varying fluorescence (x-axis) and time derivative (color axis) threshold parameters to various stimuli. Thresholds depend on the standard deviation (STD) of either fluorescence or time derivative of the 10 s window preceding the stimulus. A ‘response’ indicates that a frame t has fulfilled two criteria within 5 s of the stimulus onset: the mean fluorescence of t:t +12 frames exceeds the fluorescence threshold, and the mean time derivative of t:t+1 exceeds the time derivative threshold. A fluorescence threshold of 2 STD marks an inflection point in number of traces called ‘responses’ to pulses of buffer, such that a lower fluorescence threshold would include false positives. The time derivative threshold of 1 STD marks another inflection point such that higher thresholds begin to exclude events for all stimuli. All subsequent analyses use the thresholds of 2 STD for fluorescence and 1 STD for time derivative. The chosen time derivative threshold is useful for constraining the response timing for latency analyses rather than excluding traces based on slope or shape. Buffer: n = 297; AWA::Chr: n = 569; 11.5 nM diacetyl: n = 230; 1.15 µM diacetyl: n = 438. ( B ) Representative selection of individual AIA calcium traces deemed ‘responses’ or ‘nonresponses’ to various stimuli. Traces that are completely blue were deemed ‘nonresponses’ using the chosen threshold of 2 STD for fluorescence and 1 STD for time derivative. Traces in both blue and orange were deemed ‘responses’ at the color transition. Orange box shows a subset of traces that were deemed ‘nonresponses’ at the chosen threshold but were deemed ‘responses’ at the lower threshold of 1 STD for fluorescence and 1 STD for time derivative. ( C ) AIA ‘responses’ to various stimuli at given threshold parameters, aligned to the initiation of each response rather than stimulus onset, and averaged. Only calcium traces that were called ‘responses’ within 5 s into stimulus are included. Threshold parameters used in this study (2 STD for fluorescence; 1 STD for time derivative) are indicated by the orange box. Shading indicates ± SEM. ( D ) Rise times of AIA ‘responses’ to various stimuli and threshold parameters. Error bars show SEM. Asterisks refer to significance of one-way ANOVA with Dunnett’s multiple comparisons test, using AWA::Chrimson as the comparison stimulus. No asterisks: not significant; *: p<0.05; **: p<0.01; ***: p<0.001.

Article Snippet: We used Metamorph 7.7.6 software to control image acquisition and light pulsing in addition to rapid stimulus switching (National Instruments NI-DAQmx connected to an Automate Valvebank 8 II actuator that controls a solenoid valve), odor selection (Hamilton 8-way distribution valve), and activation of an external red LED for Chrimson stimulation (Mightex Precision LED Spot Light, 617 nm, PLS-0617–030 s; attached to Chroma ET605/50x filter to narrow band to 605 ± 25 nm).

Techniques: Fluorescence, Standard Deviation, Selection, Comparison

Journal: eLife

Article Title: Reliability of an interneuron response depends on an integrated sensory state

doi: 10.7554/eLife.50566

Figure Lengend Snippet: ( A – C ) Negative correlation between AIA GCaMP fluorescence at baseline and magnitude of responses to AWA::Chrimson ( A ), 11.5 nM diacetyl ( B ), and 1.15 µM diacetyl ( C ). Only responses to the first stimulus pulse are included in this analysis. AWA::Chrimson: n = 176; 11.5 nM diacetyl: n = 91; 1.15 µM diacetyl: n = 208. ( D – I ) Mean AIA responses to various stimuli that resulted in AIA activation within 5 s of stimulus, aligned to the beginning of AIA activation rather than stimulus time. Shading indicates ± SEM. ( D ) WT versus odr-7 and odr-10 to 1.15 µM diacetyl, shown in . ( E and F ) WT versus animals expressing a transgene encoding Tetanus Toxin Light Chain A (TeTx) in AWA to 1.15 µM diacetyl ( E ) or AWA::Chrimson ( F ), shown in and . ( G and H ) WT versus unc-7 unc-9 animals to 11.5 nM diacetyl ( G ) or AWA::Chrimson ( H ), shown in . ( I ) unc-7 mutants versus unc-7 unc-9 double mutants, unc-7 unc-9; AWA,AIA::unc-9(+) transgenic rescue animals, and unc-7 unc-9; AWA,AIA::unc-9(fc16) transgenic control animals with an inactivating point mutation, shown in . ( J – L ) Mean AIA responses to 11.5 nM diacetyl ( J ), 1.15 µM diacetyl ( K ), and AWA::Chrimson ( L ) that resulted in activation within 5 s of stimulus, aligned to the beginning of AIA activation rather than stimulus time, in WT versus unc-13 , unc-18(e234) , and unc-18(e81) animals, shown in , , and . ( M ) Rise times of AIA responses to AWA::Chrimson in WT, unc-13 , unc-18(e234) , and unc-18(e81) synaptic transmission mutants, shown in . ( N – P ) Mean AIA responses to AWA::Chrimson stimulation that resulted in activation within 5 s of stimulus, aligned to the beginning of AIA activation rather than stimulus time, in various animals lacking either glutamate release or cellular function of specific sensory neurons, shown in . ( N ) eat-4-FRT , eat-4-FRT; AWC,ASE,ASK,ASG::nFlippase , eat-4-FRT; ASG::nFlippase , eat-4-FRT; AWC,ASE::nFlippase , eat-4-FRT; ASK::nFlippase , and eat-4-FRT; AWC::nFlippase . ( O ) eat-4-FRT , unc-18(e234) , WT, and WT expressing nFlippase in AWC, ASE, ASK, and ASG. ( P ) WT and che-1 animals (ASE cell fate mutants). For ( A ), ( B ), and ( C ), asterisks refer to significance of linear regression slope differing from 0. ***: p<0.001. For ( M ), ns refers to a lack of statistical significance of one-way ANOVA with Dunnett’s multiple comparisons test. See for sample sizes and test details. For ( P ), ns refers to lack of statistical significance of an unpaired t-test. For all others, asterisks refer to statistical significance of one-way ANOVA with Dunnett’s multiple comparisons test versus unc-7 (for I only) or versus WT (for all others). ns: not significant; *: p<0.05; **: p<0.01; ***: p<0.001. See for sample sizes and test details.

Article Snippet: We used Metamorph 7.7.6 software to control image acquisition and light pulsing in addition to rapid stimulus switching (National Instruments NI-DAQmx connected to an Automate Valvebank 8 II actuator that controls a solenoid valve), odor selection (Hamilton 8-way distribution valve), and activation of an external red LED for Chrimson stimulation (Mightex Precision LED Spot Light, 617 nm, PLS-0617–030 s; attached to Chroma ET605/50x filter to narrow band to 605 ± 25 nm).

Techniques: Fluorescence, Activation Assay, Expressing, Transgenic Assay, Control, Mutagenesis, Transmission Assay, Cell Function Assay

Journal: eLife

Article Title: Reliability of an interneuron response depends on an integrated sensory state

doi: 10.7554/eLife.50566

Figure Lengend Snippet: ( A ) Cumulative response time profiles of AIA responses to AWA::Chrimson stimulation in WT versus animals expressing Tetanus Toxin Light Chain A (TeTx) in AWA. ( B and D ) Cumulative response time profiles of AIA responses to 11.5 nM diacetyl ( B ) and AWA::Chrimson stimulation ( D ) in WT versus unc-7 unc-9 animals (innexin double mutants). ( C ) Heat maps of AIA responses to 11.5 nM diacetyl in WT versus unc-7 unc-9 animals shown in ( B ). ( E ) Heat maps of AIA responses to 1.15 µM diacetyl in unc-7 mutants, unc-7 unc-9 double mutants, unc-7 unc-9; AWA,AIA::unc-9(+) transgenic rescue animals, and unc-7 unc-9; AWA,AIA::unc-9(fc16) transgenic control animals bearing an inactivating point mutation, shown in . ( F and H ) AIA ( F ) and AWA ( H ) responses to 10 s pulses of AIA::Chrimson stimulation in WT and unc-18 animals (synaptic transmission mutants); one row per calcium trace. ( G ) Cumulative response time profiles of AIA responses shown in ( F ). ( I ) Cumulative response time profiles of AWA responses shown in ( H ). Asterisks refer to Kolmogorov-Smirnov test significance versus WT over full 10 s stimulus pulse. ns: not significant; *: p<0.05; **: p<0.01. See for sample sizes and test details.

Article Snippet: We used Metamorph 7.7.6 software to control image acquisition and light pulsing in addition to rapid stimulus switching (National Instruments NI-DAQmx connected to an Automate Valvebank 8 II actuator that controls a solenoid valve), odor selection (Hamilton 8-way distribution valve), and activation of an external red LED for Chrimson stimulation (Mightex Precision LED Spot Light, 617 nm, PLS-0617–030 s; attached to Chroma ET605/50x filter to narrow band to 605 ± 25 nm).

Techniques: Expressing, Transgenic Assay, Control, Mutagenesis, Transmission Assay

Journal: eLife

Article Title: Reliability of an interneuron response depends on an integrated sensory state

doi: 10.7554/eLife.50566

Figure Lengend Snippet: ( A ) Simplified diagram of connections between AWA, AIA and four glutamatergic sensory neurons, based on . ( B ) Schematic of cell-selective glutamate knockout genetic strategy . The eat-4 locus is excised only in the presence of flippase. ORF: open reading frame; UTR: untranslated region; FRT: flippase recombinase target. ( C–H ) Cumulative response time profiles of AIA responses to AWA::Chrimson stimulation in various animals lacking either glutamate release or cellular function of specific sensory neurons. For ( D–G ), dotted black and blue lines are control and eat-4-FRT; AWC,ASE,ASK,ASG::nFlippase , respectively, from ( C ). ( C ) Control ( eat-4-FRT genetic background with no flippase expression), unc-18 , and eat-4-FRT; AWC,ASE,ASK,ASG::nFlippase animals. ( D ) eat-4-FRT; ASK::nFlippase animals. ( E ) eat-4-FRT; ASG::nFlippase animals. ( F ) eat-4-FRT; AWC::nFlippase animals. ( G ) eat-4-FRT; AWC+ASE::nFlippase animals. ( H ) WT and che-1 animals (ASE cell fate mutants). ( I ) Cumulative response time profiles of AIA responses to AWA::Chrimson stimulation in WT, unc-18 animals, unc-18; AWC,ASE::unc-18(+) transgenic rescue animals (two lines), and unc-18; AWC,ASE::unc-18(e234) transgenic control animals. Asterisks refer to Kolmogorov-Smirnov significance versus eat-4-FRT controls ( C–G ) or WT ( H, I ) over full 10 s stimulus pulse. ns: not significant; **: p<0.01; ***: p<0.001. See for sample sizes and test details. Heat maps of data from appear in . Additional representations of data from – appear in . Figure 4—source data 1. Source data for and figure supplements.

Article Snippet: We used Metamorph 7.7.6 software to control image acquisition and light pulsing in addition to rapid stimulus switching (National Instruments NI-DAQmx connected to an Automate Valvebank 8 II actuator that controls a solenoid valve), odor selection (Hamilton 8-way distribution valve), and activation of an external red LED for Chrimson stimulation (Mightex Precision LED Spot Light, 617 nm, PLS-0617–030 s; attached to Chroma ET605/50x filter to narrow band to 605 ± 25 nm).

Techniques: Knock-Out, Cell Function Assay, Control, Expressing, Transgenic Assay

Journal: eLife

Article Title: Reliability of an interneuron response depends on an integrated sensory state

doi: 10.7554/eLife.50566

Figure Lengend Snippet: ( A ) Heat maps of AIA responses to AWA::Chrimson in eat-4-FRT control strain, unc-18, FRT+nFlippase a nimals lacking glutamate release in specific sensory neurons, shown in , WT, and WT control strain expressing AWC,ASE,ASK,ASG::nFlippase. ( B ) Cumulative response time profiles of AIA responses to AWA::Chrimson stimulation in WT animals, animals with the genetically modified eat-4-FRT locus alone, or animals with the AWC,ASE,ASK,AWG::nFlippase transgene alone. All genotypes include the AWA::Chrimson transgene. ( C ) Heat maps of AIA responses to AWA::Chrimson in WT and che-1 animals (ASE cell fate mutants), shown in . ( D ) Heat maps of AIA responses to AWA::Chrimson in WT, unc-18 , unc-18; AWC,ASE::unc-18(e234) transgenic control animals bearing an inactivating point mutation, and unc-18; AWC,ASE::unc-18(+) transgenic rescue animals (two lines).

Article Snippet: We used Metamorph 7.7.6 software to control image acquisition and light pulsing in addition to rapid stimulus switching (National Instruments NI-DAQmx connected to an Automate Valvebank 8 II actuator that controls a solenoid valve), odor selection (Hamilton 8-way distribution valve), and activation of an external red LED for Chrimson stimulation (Mightex Precision LED Spot Light, 617 nm, PLS-0617–030 s; attached to Chroma ET605/50x filter to narrow band to 605 ± 25 nm).

Techniques: Control, Expressing, Genetically Modified, Transgenic Assay, Mutagenesis

Journal: eLife

Article Title: Reliability of an interneuron response depends on an integrated sensory state

doi: 10.7554/eLife.50566

Figure Lengend Snippet: ( A, C, D and E ) Cumulative response time profiles of AIA responses to 1.15 µM diacetyl in WT versus odr-7 animals (AWA cell fate mutants) ( A ), odr-10 animals (AWA diacetyl receptor mutants) ( A ), animals expressing a transgene encoding Tetanus Toxin Light Chain A (TeTx) in AWA ( C ), unc-7 or unc-9 animals (innexin mutants) ( D ), and unc-7 unc-9 double mutants ( E ). ( B ) Heat maps of AIA responses to 1.15 µM diacetyl in WT, odr-7 , and odr-10 animals from ( A ). ( F ) unc- 9 innexin rescue in AWA and AIA. Cumulative response time profiles of AIA responses to 1.15 µM diacetyl in unc-7 innexin mutants, unc-7 unc-9 double mutants, unc-7 unc-9; AWA,AIA::unc-9(+) transgenic rescue animals, and unc-7 unc-9; AWA,AIA::unc-9(fc16) transgenic control animals. ( G and I ) AIA ( G ) and AWA ( I ) responses to 10 s pulses of AIA::Chrimson stimulation in WT and unc-7 unc-9 animals; one row per calcium trace. Note that scale bar in ( I ) differs from scale bar in . ( H ) Cumulative response time profiles of AIA responses shown in ( G ). ( J ) Cumulative response time profiles of AWA responses shown in ( I ). Asterisks refer to Kolmogorov-Smirnov test significance versus WT ( A, C, D, E, H and J ) or versus unc-7 ( F ) over full 10 s stimulus pulse. ns: not significant; ***: p<0.001. See for sample sizes and test details. Additional heat maps of data from appear in . Figure 2—source data 1. Source data for and figure supplement.

Article Snippet: We used Metamorph 7.7.6 software to control image acquisition and light pulsing in addition to rapid stimulus switching (National Instruments NI-DAQmx connected to an Automate Valvebank 8 II actuator that controls a solenoid valve), odor selection (Hamilton 8-way distribution valve), and activation of an external red LED for Chrimson stimulation (Mightex Precision LED Spot Light, 617 nm, PLS-0617–030 s; attached to Chroma ET605/50x filter to narrow band to 605 ± 25 nm).

Techniques: Expressing, Transgenic Assay, Control

Journal: eLife

Article Title: Reliability of an interneuron response depends on an integrated sensory state

doi: 10.7554/eLife.50566

Figure Lengend Snippet: ( A – C ) Magnitude of individual ASK ( A ), AWC ( B ), or ASE ( C ) responses to buffer (0), 11.5 nM or 1.15 µM diacetyl shown in . Boxes show median and interquartile range. ( D ) Magnitude of individual ASH responses to buffer (0), 1.15 µM diacetyl, or 100 mM NaCl in WT animals. Boxes show median and interquartile range. ( E – G ) Cumulative response time profiles of responses from ( A ), ( B ), and ( C ). Only the first 5 s are shown. ( H ) Mean ASH responses to pulses of buffer (0), 1.15 µM diacetyl, or 100 mM NaCl in WT animals shown in ( D ). Shading indicates ± SEM. ( I – K ) Magnitude of individual ASK ( I ), AWC ( J ), or ASE ( K ) responses to pulses of 1.15 µM diacetyl in WT versus unc-18 and odr-10 animals. Boxes show median and interquartile range. ( L and M ) Mean ASK ( L ) and AWC ( M ) response to pulses of 1.15 µM diacetyl in WT versus unc-18 and odr-10 animals shown in ( I ) and ( J ). Shading indicates ± SEM. ( N ) Mean ASK, AWC, and ASE responses to pulses of AWA::Chrimson stimulation in WT animals. Shading indicates ± SEM. ( O ) Magnitude of individual responses in ( N ). Boxes show median and interquartile range. For ( A ), ( B ), ( C ), ( D ), ( I ), ( J ), and ( K ), asterisks refer to statistical significance of a one-way ANOVA with Dunnett’s multiple comparisons test versus buffer ( A–D and I–K ) or versus WT ( I–K ). ns: not significant; **: p<0.01; ***: p<0.001. For ( O ), ns refers to lack of statistical significance in paired t-tests comparing pre-light to within-light periods in same neurons. See for sample sizes and test details. For ( E ), ( F ), and ( G ), asterisks refer to Kolmogorov-Smirnov test significance versus buffer over full 10 s stimulus pulse. ns: not significant; *: p<0.05; **: p<0.01; ***: p<0.001. See for sample sizes and test details.

Article Snippet: We used Metamorph 7.7.6 software to control image acquisition and light pulsing in addition to rapid stimulus switching (National Instruments NI-DAQmx connected to an Automate Valvebank 8 II actuator that controls a solenoid valve), odor selection (Hamilton 8-way distribution valve), and activation of an external red LED for Chrimson stimulation (Mightex Precision LED Spot Light, 617 nm, PLS-0617–030 s; attached to Chroma ET605/50x filter to narrow band to 605 ± 25 nm).

Techniques:

Journal: eLife

Article Title: Reliability of an interneuron response depends on an integrated sensory state

doi: 10.7554/eLife.50566

Figure Lengend Snippet:

Article Snippet: We used Metamorph 7.7.6 software to control image acquisition and light pulsing in addition to rapid stimulus switching (National Instruments NI-DAQmx connected to an Automate Valvebank 8 II actuator that controls a solenoid valve), odor selection (Hamilton 8-way distribution valve), and activation of an external red LED for Chrimson stimulation (Mightex Precision LED Spot Light, 617 nm, PLS-0617–030 s; attached to Chroma ET605/50x filter to narrow band to 605 ± 25 nm).

Techniques: Software

Journal: European Journal of Human Genetics

Article Title: FIG4 variants in central European patients with amyotrophic lateral sclerosis: a whole-exome and targeted sequencing study

doi: 10.1038/ejhg.2016.186

Figure Lengend Snippet: Rare or novel heterozygous non-silent variants in the FIG4 gene predicted to be deleterious identified in 201 central European ALS patients

Article Snippet: Varying degrees of brain atrophy, particularly in the frontoparietal region, were found in all FIG4 variant carriers by cranial MRI. table ft1 table-wrap mode="anchored" t5 caption a7 Patient Amino-acid change Inheri-tance Gender Country of origin Age of onset (years) Site of onset Diagnosis El Escorial Diseaseduration (years) a Neurological non-motor neuron symptoms EMG NCS Cranial MRI MD072 p.(I41T) Sporadic F Italy 43 Spinal PLS Possible ALS 12.25 Pathological laughing and crying No acute/chronic denervation Normal Precentral gyrus thinning, mild frontoparietal atrophy, no ventricular enlargement, no T2 hyperintensity in the CST FamALS006–01 p.(F254Sfs*8) Familial M Germany 40 Spinal ALS Clinically probable ALS 2.67 None Acute/chronic denervation UE, LE, thoracic Reduction of CMAP and prolongation of DML Severe frontoparietal atrophy, no ventricular enlargement, no T2 hyperintensity in the CST VALS042 p.(D307N) Sporadic M Germany 78 Bulbar ALS-UMN Clinically probable ALS 5.25 Sensory impairment for vibration, light touch Chronic denervation UE, LE Motor-sensory axonal neuropathy Age-related symmetric atrophy and ventricular enlargement, no T2 hyperintensity in the CST VALS007 p.(T540I) Sporadic F Germany 72 Bulbar PLS Possible ALS 3.25 Mild cognitive impairment, sensory impairment for vibration Sparse and stable signs of acute/chronic denervation (LE only) Reduction of CMAP and prolongation of DML Moderate frontoparietal atrophy and ventricular enlargement, no T2 hyperintensity in the CST VALS012 p.(Y647C) Sporadic M Germany 66 Spinal ALS-flail arm — 5.25 None Acute/chronic denervation UE, LE Motor-sensory axonal neuropathy Mild frontoparietal atrophy and ventricular enlargement, no T2 hyperintensity in the CST VALS015 p.(S853L) Sporadic F Germany 48 Spinal ALS Clinically probable ALS 0.92 Minimal sensory impairment for vibration Acute/chronic denervation Mild motor–sensory axonal neuropathy Mild frontoparietal atrophy, no ventricular enlargement, no T2 hyperintensity in the CST Open in a separate window Abbreviations: ALS, amyotrophic lateral sclerosis; CMAP, compound motor action potential; CST, corticospinal tract; DML, distal motor latency; EMG, electromyography; F, female; LE, lower extremity; M, male; MRI, magnetic resonance imaging; NCS, nerve conduction study; PLS, primary lateral sclerosis; UE, upper extremity; UMN, upper motor neuron.

Techniques:

Journal: European Journal of Human Genetics

Article Title: FIG4 variants in central European patients with amyotrophic lateral sclerosis: a whole-exome and targeted sequencing study

doi: 10.1038/ejhg.2016.186

Figure Lengend Snippet: Clinical, electrophysiological, and neuroradiological characteristics of ALS patients carrying heterozygous deleterious FIG4 variants

Article Snippet: Varying degrees of brain atrophy, particularly in the frontoparietal region, were found in all FIG4 variant carriers by cranial MRI. table ft1 table-wrap mode="anchored" t5 caption a7 Patient Amino-acid change Inheri-tance Gender Country of origin Age of onset (years) Site of onset Diagnosis El Escorial Diseaseduration (years) a Neurological non-motor neuron symptoms EMG NCS Cranial MRI MD072 p.(I41T) Sporadic F Italy 43 Spinal PLS Possible ALS 12.25 Pathological laughing and crying No acute/chronic denervation Normal Precentral gyrus thinning, mild frontoparietal atrophy, no ventricular enlargement, no T2 hyperintensity in the CST FamALS006–01 p.(F254Sfs*8) Familial M Germany 40 Spinal ALS Clinically probable ALS 2.67 None Acute/chronic denervation UE, LE, thoracic Reduction of CMAP and prolongation of DML Severe frontoparietal atrophy, no ventricular enlargement, no T2 hyperintensity in the CST VALS042 p.(D307N) Sporadic M Germany 78 Bulbar ALS-UMN Clinically probable ALS 5.25 Sensory impairment for vibration, light touch Chronic denervation UE, LE Motor-sensory axonal neuropathy Age-related symmetric atrophy and ventricular enlargement, no T2 hyperintensity in the CST VALS007 p.(T540I) Sporadic F Germany 72 Bulbar PLS Possible ALS 3.25 Mild cognitive impairment, sensory impairment for vibration Sparse and stable signs of acute/chronic denervation (LE only) Reduction of CMAP and prolongation of DML Moderate frontoparietal atrophy and ventricular enlargement, no T2 hyperintensity in the CST VALS012 p.(Y647C) Sporadic M Germany 66 Spinal ALS-flail arm — 5.25 None Acute/chronic denervation UE, LE Motor-sensory axonal neuropathy Mild frontoparietal atrophy and ventricular enlargement, no T2 hyperintensity in the CST VALS015 p.(S853L) Sporadic F Germany 48 Spinal ALS Clinically probable ALS 0.92 Minimal sensory impairment for vibration Acute/chronic denervation Mild motor–sensory axonal neuropathy Mild frontoparietal atrophy, no ventricular enlargement, no T2 hyperintensity in the CST Open in a separate window Abbreviations: ALS, amyotrophic lateral sclerosis; CMAP, compound motor action potential; CST, corticospinal tract; DML, distal motor latency; EMG, electromyography; F, female; LE, lower extremity; M, male; MRI, magnetic resonance imaging; NCS, nerve conduction study; PLS, primary lateral sclerosis; UE, upper extremity; UMN, upper motor neuron.

Techniques:

Journal: Journal of Clinical Medicine

Article Title: Breath Analysis via Gas Chromatography–Mass Spectrometry (GC-MS) in Chronic Coronary Syndrome (CCS): A Proof-of-Concept Study

doi: 10.3390/jcm13195857

Figure Lengend Snippet: Exhaled breath collection and analysis. Each patient provided a single breath sample into a Pneumopipe ® device containing an adsorbing cartridge. Volatile organic compounds (VOCs) from Tenax cartridges were collected in 500 mL Tedlar bags and analyzed using the carboxen solid phase microextraction (SPME) gas chromatography–mass spectrometry (GC-MS) technique. The SPME fiber was thermally desorbed in the GC injector at 280 °C. Data were derived from GC-MS chromatogram analysis, expressed as the ratio of the VOC area to the reference peak area, and analyzed using a partial least square discriminant analysis (PLS-DA) model.

Article Snippet: PLS-DA was performed using the PLS Toolbox software ( https://www.mdpi.com/1424-8220/21/13/4464 , https://www.mdpi.com/2072-4292/13/20/4026 , both accessed on 25 September 2024) (Eigenvector Research Inc., Wenatchee, WA, USA) in the MATLAB environment (The MathWorks Inc., Natick, MA, USA).

Techniques: Solid-phase Microextraction, Gas Chromatography, Mass Spectrometry, Gas Chromatography-Mass Spectrometry, Derivative Assay

Journal: Journal of Clinical Medicine

Article Title: Breath Analysis via Gas Chromatography–Mass Spectrometry (GC-MS) in Chronic Coronary Syndrome (CCS): A Proof-of-Concept Study

doi: 10.3390/jcm13195857

Figure Lengend Snippet: Confusion matrix of the partial least square discriminant analysis (PLS-DA) model among patients with and without the indication of myocardial revascularization.

Article Snippet: PLS-DA was performed using the PLS Toolbox software ( https://www.mdpi.com/1424-8220/21/13/4464 , https://www.mdpi.com/2072-4292/13/20/4026 , both accessed on 25 September 2024) (Eigenvector Research Inc., Wenatchee, WA, USA) in the MATLAB environment (The MathWorks Inc., Natick, MA, USA).

Techniques: Gas Chromatography-Mass Spectrometry

Journal: Journal of Clinical Medicine

Article Title: Breath Analysis via Gas Chromatography–Mass Spectrometry (GC-MS) in Chronic Coronary Syndrome (CCS): A Proof-of-Concept Study

doi: 10.3390/jcm13195857

Figure Lengend Snippet: Confusion matrix of the partial least square discriminant analysis (PLS-DA) model in the subgroup analysis (Group 1 vs. Group 2, 3 vs. Group 4, 5).

Article Snippet: PLS-DA was performed using the PLS Toolbox software ( https://www.mdpi.com/1424-8220/21/13/4464 , https://www.mdpi.com/2072-4292/13/20/4026 , both accessed on 25 September 2024) (Eigenvector Research Inc., Wenatchee, WA, USA) in the MATLAB environment (The MathWorks Inc., Natick, MA, USA).

Techniques: Gas Chromatography-Mass Spectrometry