This dataset contains the analysis-ready raw measurements and GraphPad Prism statistical analysis files associated with the manuscript "Integrin-Talin axis regulates mechanosensory synapse development and behavioral habituation to mechanical stimulation" authored by Jonathan Amezquita, Nelson Ayala, Muriel Desbois, Karla Opperman, Neal Mathew, and Brock Grill.

The dataset is hosted by Dryad under DOI: 10.5061/dryad.msbcc2g8w.

We have submitted GraphPad Prism project files (.prism) corresponding to the quantitative analyses reported in the manuscript. Each Prism file contains the numerical values used for statistical analysis, the organization of experimental groups, and the associated statistical analysis for the indicated figure panel(s). The deposited data include measurements of C. elegans PLM mechanosensory neuron presynaptic branch defects, presynaptic bouton development, fluorescence intensity measurements, and behavioral habituation to repeated mechanical stimulation.

The deposited Prism files contain the analysis-ready measurements used to generate the reported statistical comparisons.

Descriptions

Figure_2D_Datafile_Final.prism

GraphPad Prism file for Figure 2D, which quantifies presynaptic branch defects in PLM mechanosensory neurons of young adult animals.

• Analysis: PLM presynaptic branch defects after cell-specific degradation of components of the PAT-3/UNC-112/TLN-1 axis and in tln-1(ok1648) hypomorphic mutants.
• Experimental system: C. elegans PLM mechanosensory neurons visualized using Pmec-7::mRFP (jsIs973).
• Experimental unit: Individual scoring count. Each count contains 20 or more PLM neurons; at least five counts were collected for each genotype from at least three independent biological experiments.
• Measured value: Percentage of PLM neurons with presynaptic branch defects.
• Units: Percent branch defects (%).
• Relevant genotype/condition labels: Control animals; PAT-3::GFP + mecDEG; UNC-112::GFP + mecDEG; GFP::TLN-1 + mecDEG; tln-1(ok1648), and any additional labels shown in the Prism file.
• Statistical analysis: Two-way ANOVA with Bonferroni correction for multiple comparisons where applicable. Significance was defined as p < 0.05.

Figure_2H-I_Datafile_Final.prism

GraphPad Prism file for Figure 2H-I, which quantifies fluorescence intensity in PLM soma regions of interest.

• Analysis: Fluorescence intensity measurements used to assess GFP::TLN-1 depletion in PLM soma, with Pmec-7::mRFP fluorescence quantified as a negative/control reporter.
• Experimental system: PLM mechanosensory neurons in young adult animals.
• Experimental unit: Individual PLM neuron/animal. Images from 10-12 PLM neurons were analyzed for each genotype across three independent biological experiments.
• Measured values: Mean fluorescence intensity in manually drawn PLM soma regions of interest from maximum-intensity projections.
• Units: Arbitrary fluorescence intensity units, as reported by the image-analysis workflow and entered into Prism.
• ROI definition: PLM soma ROIs were manually drawn in Zeiss Zen software using the spline tool.
• Figure 2H: GFP::TLN-1 fluorescence intensity in PLM soma.
• Figure 2I: Pmec-7::mRFP fluorescence intensity in PLM soma.
• Statistical analysis: Unpaired, two-tailed Student's t-test. Significance was defined as p < 0.05.

Figure_2J-K_Datafile_Final.prism

GraphPad Prism file for Figure 2J-K, which quantifies fluorescence intensity in PLM proximal axon regions of interest.

• Analysis: Fluorescence intensity measurements used to assess GFP::TLN-1 depletion in the PLM proximal axon, with Pmec-7::mRFP fluorescence quantified as a negative/control reporter.
• Experimental system: PLM mechanosensory neurons in young adult animals.
• Experimental unit: Individual PLM neuron/animal. Images from 10-12 PLM neurons were analyzed for each genotype across three independent biological experiments.
• Measured values: Mean fluorescence intensity in PLM proximal axon regions of interest from maximum-intensity projections.
• Units: Arbitrary fluorescence intensity units, as reported by the image-analysis workflow and entered into Prism.
• ROI definition: Proximal axon ROIs were defined using 4.5 µm² boxes in Zeiss Zen software.
• Figure 2J: GFP::TLN-1 fluorescence intensity in PLM proximal axon.
• Figure 2K: Pmec-7::mRFP fluorescence intensity in PLM proximal axon.
• Statistical analysis: Unpaired, two-tailed Student's t-test. Significance was defined as p < 0.05.

Figure_3E_Datafile_Final.prism

GraphPad Prism file for Figure 3E, which quantifies PLM presynaptic branch defects caused by transgenic expression of wild-type and constitutively active TLN-1 variants.

• Analysis: PLM presynaptic branch defects after pan-neuronal expression of TLN-1 or constitutively active TLN-1 point mutants.
• Experimental system: PLM mechanosensory neurons visualized using Pmec-7::mRFP (jsIs973).
• Experimental unit: Individual scoring count. Data were collected and averaged from 2-3 transgenic lines for each genotype. At least five counts were collected for each genotype, with 20 or more animals per count.
• Measured value: Percentage of PLM neurons with presynaptic branch defects.
• Units: Percent branch defects (%).
• Relevant genotype/condition labels: Control animals; pan-neuronal Prgef-1::TLN-1 expression; constitutively active TLN-1 T1767E; constitutively active TLN-1 E1770A, and any additional labels shown in the Prism file.
• Interpretation key: TLN-1 T1767E and TLN-1 E1770A are point mutations that disrupt TLN-1 autoinhibition and promote constitutively active Talin signaling.
• Statistical analysis: Two-way ANOVA with Bonferroni correction. Significance was defined as p < 0.05.

Figure_4C_Datafile_Final.prism

GraphPad Prism file for Figure 4C, which quantifies genetic interactions between the PAT-3/UNC-112/TLN-1 axis and RPM-1 in PLM presynaptic branch development.

• Analysis: PLM presynaptic branch defects in rpm-1 loss-of-function mutants, in mecDEG-mediated degradation conditions affecting the PAT-3/UNC-112/TLN-1 axis, and in combined rpm-1 mutant/degradation backgrounds.
• Experimental system: PLM mechanosensory neurons visualized using Pmec-7::mRFP (jsIs973).
• Experimental unit: Individual scoring count. At least five counts were collected for each genotype, with 20 or more animals per count.
• Measured value: Percentage of PLM neurons with presynaptic branch defects.
• Units: Percent branch defects (%).
• Relevant genotype/condition labels: Control animals; rpm-1(lf); PAT-3::GFP + mecDEG; UNC-112::GFP + mecDEG; GFP::TLN-1 + mecDEG; rpm-1; PAT-3::GFP + mecDEG; rpm-1; UNC-112::GFP + mecDEG; rpm-1; GFP::TLN-1 + mecDEG, and any additional labels shown in the Prism file.
• Statistical analysis: Two-way ANOVA with Bonferroni correction for multiple comparisons. Significance was defined as p < 0.05.

Figure_5C-E_Datafile_Final.prism

GraphPad Prism file for Figure 5C-E, which contains developmental time-course data for PLM presynaptic bouton development and maintenance.

• Analysis: Developmental time-course analysis of presynaptic bouton formation and maintenance in PLM neurons.
• Experimental system: PLM mechanosensory neurons visualized using Pmec-7::mRFP (jsIs973).
• Experimental unit: Individual PLM neuron scored for the presence or absence of presynaptic boutons.
• Measured values: Frequency or percentage of PLM neurons with or lacking presynaptic boutons at the indicated developmental time points.
• Units: Percent of PLM neurons in the indicated phenotypic category (%), or counts of PLM neurons in each category as organized in the Prism file.
• Developmental time points: 6 h, 16 h, and 48 h post-hatch (PH), as indicated in the Prism file and figure panel.
• Sample structure: 50-60 PLM neurons were scored for each time point and genotype from three or more independent imaging sessions.
• Relevant genotype/condition labels: Control animals; rpm-1(lf); GFP::TLN-1 + mecDEG; rpm-1; GFP::TLN-1 + mecDEG, and any additional labels shown in the Prism file.
• Statistical analysis: Fisher's exact test. Significance was defined as p < 0.05.

Figure_6C-E_Datafile_Final.prism

GraphPad Prism file for Figure 6C-E, which contains behavioral habituation data from repeated mechanical stimulation assays.

• Analysis: Behavioral habituation to repeated mechanical stimulation in control and GFP::TLN-1 degradation conditions.
• Experimental system: C. elegans populations assessed using Multi-Worm Tracker (MWT). Pmec-7::mRFP (jsIs973) was used as a control marker to label mechanosensory neurons for branch phenotype verification prior to behavioral testing.
• Stimulus protocol: 45 mechanical tap stimuli delivered at 10 s interstimulus intervals.
• Experimental unit: Plate-level behavioral recording. Ten plates were recorded for each genotype, with approximately 50-100 animals per plate. Data were collected across five independent biological experiments performed on different days, with two plates per genotype per day.
• Figure 6C measured value: Mean reversal probability at each tap event across the habituation protocol.
• Figure 6D measured value: Initial touch response, calculated from the first tap event.
• Figure 6E measured value: Final habituation level/asymptote, as represented in the Prism file.
• Units: Reversal probability or response probability, as plotted in the Prism file.
• Relevant genotype/condition labels: Control animals; GFP::TLN-1 + mecDEG, and any additional labels shown in the Prism file.
• Exclusion criteria: Individual tracking events were excluded when MWT acquisition failed because the tapper failed to fire or animal tracking was momentarily lost.
• Statistical analysis: Two-way ANOVA for habituation profiles. Initial tap response and final habituation level were analyzed using two-way ANOVA with Bonferroni correction for multiple comparisons. Significance was defined as p < 0.05.

Common variables, labels, and interpretation keys

The Prism files are organized by genotype or experimental condition, figure panel, and measured outcome. Exact table organization varies by Prism file because the dataset includes column-style analyses, grouped analyses, developmental time-course analyses, and behavioral habituation profiles.

• Genotype/condition: Experimental group or genetic background shown in the Prism file. Labels correspond to the groups plotted in the associated manuscript figure panel.
• Control: Reference genotype or transgenic control used for the associated comparison.
• PLM neuron: Posterior lateral mechanosensory neuron in C. elegans.
• Presynaptic branch: Collateral PLM branch that contains presynaptic specializations/boutons.
• Presynaptic branch defect: Abnormal or missing PLM presynaptic branch phenotype scored in the associated analysis.
• Presynaptic bouton: Presynaptic specialization observed along the PLM presynaptic branch.
• Presynaptic bouton frequency: Frequency or percentage of PLM neurons with presynaptic boutons at the indicated developmental time point.
• Post-hatch (PH): Time after hatching. Developmental time-course analyses include 6 h, 16 h, and 48 h PH animals.
• Fluorescence intensity: Mean fluorescence value measured within the indicated PLM soma or proximal axon region of interest. Values are reported in arbitrary fluorescence intensity units unless otherwise specified in the Prism file.
• ROI: Region of interest used for fluorescence quantification.
• Reversal probability: Probability or fraction of animals that reversed following a tap stimulus in the behavioral habituation assay.
• Initial tap response: Behavioral response measured at the first tap event.
• Final habituation level/asymptote: Late-stage response level used to quantify the final habituated response, as represented in the Prism file.
• Biological experiment: Independent experiment performed on a separate day or independent imaging/behavioral session, as applicable to the assay.
• Count: For branch-defect analyses, one count refers to a scoring set of 20 or more PLM neurons for a genotype or condition.
• SEM: Standard error of the mean.
• ns: Not significant.
• p-value significance notation: Significance thresholds in the manuscript are indicated as follows: ****p < 0.0001, ***p < 0.001, **p < 0.01, *p < 0.05, ns = not significant.

Genotype, transgene, and abbreviation key

• C. elegans: C. elegans, the nematode model organism used for all experiments in this dataset.
• PAT-3: C. elegans beta-integrin.
• UNC-112: C. elegans kindlin-family protein.
• TLN-1: C. elegans talin.
• RPM-1: C. elegans PHR-family ubiquitin ligase that regulates neuronal development.
• mecDEG: CRISPR-based cell-specific degradation system used to impair GFP-tagged proteins in mechanosensory neurons.
• PAT-3::GFP + mecDEG: Condition in which GFP-tagged PAT-3 is targeted by mecDEG in mechanosensory neurons.
• UNC-112::GFP + mecDEG: Condition in which GFP-tagged UNC-112 is targeted by mecDEG in mechanosensory neurons.
• GFP::TLN-1 + mecDEG: Condition in which GFP-tagged TLN-1 is targeted by mecDEG in mechanosensory neurons.
• tln-1(ok1648): Hypomorphic tln-1 mutant condition.
• rpm-1(ju44): rpm-1 loss-of-function mutant condition.
• Pmec-7::mRFP (jsIs973): Mechanosensory neuron marker used to visualize PLM neurons and PLM presynaptic branch morphology.
• Prgef-1: Pan-neuronal promoter used for transgenic TLN-1 expression in the Figure 3 analysis.
• TLN-1 T1767E: Constitutively active TLN-1 point-mutant condition used to disrupt TLN-1 autoinhibition.
• TLN-1 E1770A: Constitutively active TLN-1 point-mutant condition used to disrupt TLN-1 autoinhibition.
• MWT: Multi-Worm Tracker, used to quantify behavioral responses to repeated mechanical stimulation.

Missing data and exclusions

For behavioral habituation analyses, individual tracking events were excluded when they were not acquired properly by MWT because the tapper failed to fire or animal tracking was momentarily lost.

Key information sources

Experimental descriptions, figure-panel assignments, sample-size definitions, and statistical analysis details are derived from the associated manuscript and figure legends.

Code/Software

GraphPad Prism is required to open the submitted .prism files. The analyses were performed using GraphPad Prism version 11.0.2. Each Prism file contains the analysis-ready numerical measurements and statistical analyses for the corresponding figure panel(s).

Zeiss Zen software version 3.9.2 was used to draw fluorescence regions of interest and quantify GFP::TLN-1 and Pmec-7::mRFP fluorescence intensity measurements in PLM neurons.

Multi-Worm Tracker (MWT) was used to collect behavioral habituation measurements in response to repeated mechanical tap stimulation.

No standalone analysis scripts are included in this Dryad deposit.

Legal and ethical considerations

All data were generated from laboratory studies of C. elegans. The dataset contains no human-subject data.