Journal: iScience

Article Title: Mrgprb4-lineage neurons indispensable in pressure induced pleasant sensation are polymodal

doi: 10.1016/j.isci.2025.111940

Figure Lengend Snippet:

Article Snippet: Mechanical stimulation device , BIOSEB SAS , Cat#BIO-RP-M.

Techniques: Virus, Recombinant, RNAscope, Software, Microscopy, Confocal Microscopy, Conditioned Place Preference

Journal: Nature

Article Title: Increased CSF drainage by non-invasive manipulation of cervical lymphatics

doi: 10.1038/s41586-025-09052-5

Figure Lengend Snippet: a , Drawing and photograph of the precision force-regulated mechano-stimulator consisting of an amplifier, force sensor, handle, shaft connecting the tip to the force sensor, and replaceable tip. The replaceable tip was an oval-shaped cotton ball, with a major axis of 1 cm and minor axis of 0.5 cm, that was attached to a 1-cm long rod securely connected to the shaft. The length of the handle was 9 cm. The force sensor made of silicone and conductive fabric was connected to the replaceable tip through the shaft. The measured force was transmitted to an amplifier connected to a personal computer. b , Graphs comparing low-magnitude (0.01-0.02 kgf) and high-magnitude (0.04-0.08 kgf) forces generated by the mechanical stimulator every 2 s over 20 s. c , Drawing (left) showing the three stimulation paths (numbered brown dashed arrows) in three regions of intact skin of mice along the course of superficial cervical lymphatics scLV-1 and scLV-2 to promote CSF flow toward the submandibular lymph node (smLN). Region 1 was from the periorbital area to the mandible. Region 2 was from the nasal sidewall to the mandible. Region 3 was along the paths of scLV-1 and scLV-2 to the submandibular lymph node. Each 1-min session of mechanical stimulation consisted of two sequences of 10 two-second strokes each, with a 20-sec rest period after the two sequences. Each sequence included four two-second strokes in Region 1, four in Region 2, and two in Region 3 (right).

Article Snippet: Treatments consisted of five 1-min sessions of mechanical stimulation for tracer studies from the SAS to scLV-1 and scLV-2 (Fig. ).

Techniques: Generated, Sequencing

Journal: Nature

Article Title: Increased CSF drainage by non-invasive manipulation of cervical lymphatics

doi: 10.1038/s41586-025-09052-5

Figure Lengend Snippet: a , Sequence of intracisternal infusion of TMR–dextran into Prox1 –GFP mice followed by mechanical stimulation at 30 min for 5 min, then imaging of TMR–dextran in scLV-1 and scLV-2 at 35 min. b , c , Fluorescence images and measurements of TMR–dextran in scLV-1 and scLV-2. Sham compared with mechanical stimulation ( b ). Anatomical positions are indicated. Scale bars, 200 μm. Each dot represents combined TMR–dextran fluorescence intensity in scLV-1 and scLV-2 from one mouse ( c ). n = 7 (sham), n = 10 (low magnitude) and n = 6 (high magnitude) from three independent experiments. The error bars indicate mean ± s.e.m. P values were calculated by Brown–Forsythe analysis of variance (ANOVA) test followed by two-tailed Dunnett’s T3 multiple comparison post-hoc test. d , Sequence of intracisternal infusion of TMR–dextran into Prox1 –GFP mice followed by mechanical stimulation at 10 min for 20 min, then imaging of TMR–dextran in the smLN at 30 min. e , f , Fluorescence images ( e ) and measurements ( f ) of TMR–dextran in the smLN of the stimulated side (top) versus unstimulated side (bottom). Scale bars, 500 μm. Each dot is the value for one mouse. n = 12 (sham), n = 10 (low magnitude) and n = 13 (high magnitude) from three independent experiments. The error bars indicate mean ± s.e.m. P values were calculated by Brown–Forsythe ANOVA test followed by two-tailed Dunnett’s T3 multiple comparison post-hoc test. g , h , Sequence of intracerebroventricular infusion of TMR–dextran into Prox1 –GFP mice followed by low-magnitude mechanical stimulation (LMMS) at 10 min for 10 min and removal of CSF at 30 min ( g ). Fluorescence in CSF after LMMS or sham is also shown ( h ). Each dot is the value for one mouse. n = 6 (sham) and n = 7 (LMMS) from three independent experiments. The error bars indicate mean ± s.e.m. P values were calculated by two-tailed unpaired t -test with Welch’s correction. FI, fluorescence intensity.

Article Snippet: Treatments consisted of five 1-min sessions of mechanical stimulation for tracer studies from the SAS to scLV-1 and scLV-2 (Fig. ).

Techniques: Sequencing, Imaging, Fluorescence, Two Tailed Test, Comparison

Journal: Nature

Article Title: Increased CSF drainage by non-invasive manipulation of cervical lymphatics

doi: 10.1038/s41586-025-09052-5

Figure Lengend Snippet: a , Sequence of surgical exposure of superficial cervical lymphatics scLV-1 in Prox1 -GFP mice, a 20-min stabilization period, and intravital imaging during and 5 min after 1-min of low-magnitude mechanical stimulation (LMMS) of the face and neck as in Extended Data Fig. . Measurements were made at 1-min intervals for 5 min after stimulation. b , Drawing showing the relative locations of region 3 of LMMS of scLV-1 and scLV-2 and the downstream window for intravital imaging (blue dashed box). c , Intravital images of scLV-1 of Prox1 -GFP mouse downstream before and after one session of LMMS. White dashed lines outline the vessel border before LMMS. Red arrowheads mark regions of TMR-dextran fluorescence. Scale bars, 20 μm. Anatomical positions are indicated in the top left corner: A, anterior; P, posterior. d , Measurements of temporal changes in TMR-dextran fluorescence, as an index of CSF outflow, and five parameters of spontaneous contraction of scLV-1 at the onset and 1-5 min after a 1-min session of LMMS (MS). At 1-min, TMR-dextran values are more than double the onset and continue to be significantly greater throughout the 5-min monitoring period. Other values show a small increase in scLV-1 mean diameter and transient increase in amplitude of spontaneous contractions but no significant change in ejection fraction, contraction frequency, or fractional pump flow. All values are expressed as % of mean baseline fluorescence before LMMS. Each dot is the mean value for n = 9 (sham), n = 7 (LMMS) mice of both sexes from three independent experiments. Error bars indicate s.e.m. P values calculated by two-way repeated-measures ANOVA followed by Sidak’s multiple comparison test. e , Sequence of repeated low-magnitude mechanical stimulation (LMMS, 20 sessions/day) for 4 days followed on day 5 by surgical exposure and intravital imaging of superficial cervical lymphatic scLV-1 in adult (8–12 weeks) Prox1 -GFP mice. Intravital imaging began after a 30-min stabilization period and lasted 3 min. f , Measurements show no difference between the sham and stimulated group in values for mean diameter, spontaneous contraction amplitude, ejection fraction, frequency, or fractional pump flow in scLV-1. Each dot is the value for one mouse. n = 9 (sham), n = 8 (LMMS) mice per group in three independent experiments. Error bars indicate mean ± s.e.m. a.u., arbitrary unit. Intravital imaging values were averaged over 3 min for each mouse. P values calculated by two-tailed Welch’s t test. g , Sequence of repeated LMMS (20 sessions/day) for 4 days followed by surgical exposure, intracisternal infusion of TMR-dextran, intravital imaging of scLV-1, low-magnitude mechanical stimulation (1 session) in adult Prox1 -GFP mice, and measurement of TMR-dextran fluorescence in scLV-1 within the imaging window. Intravital imaging began after a 30-min stabilization period and lasted 5 min after LMMS (1 session) or sham. h , Comparison of three conditions showing significantly greater TMR-dextran fluorescence in scLV-1 in the repeated stimulation group than in the sham group. The magnitude of TMR-dextran fluorescence in the scLV-1 after repeated stimulation over 4 days did not differ from corresponding values after one stimulation session (data from Extended Data Fig. 14d). Each dot is the value for n = 9 (sham), n = 7 (1 session LMMS or repeated LMMS) mice of both sexes from three independent experiments. Error bars indicate mean ± s.e.m. a.u., arbitrary unit. P values calculated by two-way repeated-measures ANOVA.

Article Snippet: Treatments consisted of five 1-min sessions of mechanical stimulation for tracer studies from the SAS to scLV-1 and scLV-2 (Fig. ).

Techniques: Sequencing, Imaging, Fluorescence, Comparison, Two Tailed Test

Journal: Nature

Article Title: Increased CSF drainage by non-invasive manipulation of cervical lymphatics

doi: 10.1038/s41586-025-09052-5

Figure Lengend Snippet: a , Sequence of intraperitoneal (i.p.) injection of PBS or l -NAME (1 mg/kg body weight) followed 120 min later by intravital imaging and measurements of superficial cervical lymphatic vessel scLV-1 of Prox1 -GFP mice (8–12 weeks old). b , Intravital image showing scLV-1 of Prox1 -GFP mouse at 120 min after treatment with PBS or l -NAME. Basal tone (smaller diameter) was greater after l -NAME. Scale bars, 30 μm. A, anterior; P, posterior anatomical position. Representative of n = 4-5 mice from three independent experiments. c , Comparison of effect of PBS or l -NAME on scLV-1 mean diameter, amplitude, frequency, ejection fraction, and fractional pump flow. Each dot is the value for one mouse. n = 5 (PBS), n = 4 ( l -NAME) mice of both sexes per group from three independent experiments. Error bars indicate mean ± s.e.m. a.u., arbitrary unit. P values calculated by two-tailed Mann-Whitney U -test. d , Sequence of PBS or l -NAME injected intraperitoneally (i.p.) followed 90 min later by intracisternal (i.c.) infusion of 1.0 μl TMR-dextran over 1 min into adult Prox1 -GFP mice followed by measurement of TMR-dextran fluorescence in the submandibular lymph node 30 min later. Low-magnitude mechanical stimulation was applied for 20 min beginning 10 min after the intracisternal infusion. e , f , Fluorescence images and measurements of TMR-dextran fluorescence in the submandibular lymph node 30 min after the intracisternal infusion and mechanical stimulation shown in d . Scale bars, 500 μm. Fluorescence in lymph node of PBS controls compared to l -NAME treatment without stimulation (sham) or after 20 low-magnitude stimulation sessions over 20 min beginning 10 min after TMR-dextran infusion. Each dot is the value for one mouse. n = 17 (PBS, sham), n = 14 (PBS, stimulated), n = 11 ( l -NAME, sham), n = 9 ( l -NAME, stimulated) mice per group from four independent experiments. Error bars indicate mean ± s.e.m. a.u., arbitrary unit. P values calculated by Brown-Forsythe ANOVA test followed by Dunnett’s T3 multiple comparison test.

Article Snippet: Treatments consisted of five 1-min sessions of mechanical stimulation for tracer studies from the SAS to scLV-1 and scLV-2 (Fig. ).

Techniques: Sequencing, Injection, Imaging, Comparison, Two Tailed Test, MANN-WHITNEY, Fluorescence

Journal: Nature

Article Title: Increased CSF drainage by non-invasive manipulation of cervical lymphatics

doi: 10.1038/s41586-025-09052-5

Figure Lengend Snippet: a , Sequence of intracisternal infusion of 1.0 μl TMR–dextran over 1 min into aged Prox1 –GFP mice (87–105 weeks of age) followed 10 min later by sham or low-magnitude mechanical stimulation of intact skin over 20 min beginning 10 min after TMR–dextran infusion. TMR–dextran fluorescence was imaged and measured in the ipsilateral smLN at 30 min. b , c , Fluorescence images ( b ) and measurements ( c ) of TMR–dextran fluorescence in the smLN after ipsilateral low-magnitude mechanical stimulation of the skin or no stimulation (sham) over 20 min. Scale bars, 500 μm. Each dot is the TMR–dextran fluorescence intensity of the smLN in one mouse. n = 6 mice per group from three independent experiments. The error bars indicate mean ± s.e.m. P values were calculated by two-tailed Mann–Whitney U -test. d , Sequence of intracisternal infusion of 1.0 μl TMR–dextran over 1 min into aged Prox1 –GFP mice (87–105 weeks of age) followed by sham or low-magnitude mechanical stimulation of intact skin over 5 min beginning 30 min after TMR–dextran infusion. TMR–dextran fluorescence was imaged and measured in ipsilateral scLV-1 and scLV-2 at 35 min. e , f , Fluorescence images and measurements of TMR–dextran fluorescence in scLV-1 and scLV-2 at 35 min after ipsilateral low-magnitude mechanical stimulation of the skin or no stimulation (sham) over 5 min. Anatomical positions are shown in the bottom left corner. Scale bars, 200 μm. Each dot is the value of the combined TMR–dextran fluorescence intensity of scLV-1 and scLV-2 from one mouse. n = 5 (sham) and n = 4 (low-magnitude mechanical stimulation) mice per group from three independent experiments. The error bars indicate mean ± s.e.m. P values were calculated by two-tailed Mann–Whitney U -test.

Article Snippet: Treatments consisted of five 1-min sessions of mechanical stimulation for tracer studies from the SAS to scLV-1 and scLV-2 (Fig. ).

Techniques: Sequencing, Fluorescence, Two Tailed Test, MANN-WHITNEY

Journal: Frontiers in Cell and Developmental Biology

Article Title: Naturally Engineered Maturation of Cardiomyocytes

doi: 10.3389/fcell.2017.00050

Figure Lengend Snippet: Large divide between natural cardiomyocyte development and in vitro PSC-CM development . PSC-CMs generally remain within the early to late fetal CM stages even after applying natural engineering approaches, such as mechanical stimulation, electrical stimulation, non-cardiomyocyte interactions, or extracellular matrix interactions to improve their overall maturity to a late PSC-CM stage.

Article Snippet: , Biomimetic pressure gradient mechanical stimulation system for mimicking cardiac cycle with fluid flow , • Duration: 3 days • 50 mmHg pressure • 10% biaxial stretch • Gradual vs immediate physiological strain , HIPS-CMs on collagen/matrigel coated flexible PDMS membrane , • N/A , • Mimics physiological cardiac cycle • Cell culture chamber performs pumping action to move fluid , • Cells cultured on surface of 3D gels , Rogers et al., .

Techniques: In Vitro

Journal: Frontiers in Cell and Developmental Biology

Article Title: Naturally Engineered Maturation of Cardiomyocytes

doi: 10.3389/fcell.2017.00050

Figure Lengend Snippet: Natural engineering approaches schematic . Mechanical stimulation, electrical stimulation, extracellular matrix interactions, and non-cardiomyocyte interactions have been utilized to mimic the natural physiological conditions of CMs during development, known as natural engineering, in order to improve PSC-CM maturity. Mechanical stimulation in the form of pulsatile flow, static and cyclic stretch mechanisms have increased engineered heart tissue (EHT) maturity. Applying an electrical stimulus to PSC-CMs and modulating the frequency, pulse duration, and field strength has led to some functional and structural PSC-CM maturation improvements. By using natural inspired extracellular matrix interactions, such as alignment, ECM composition, and stiffness changes, researchers have been able to increase PSC-CM maturity. Non-cardiomyocyte interactions with PSC-CMs have also led to increased maturation. Overall, these four main natural engineering approaches occur simultaneously and synergistically during the natural developmental paradigm in vivo and therefore need to be recapitulated in vitro to further improve PSC-CM maturity.

Article Snippet: , Biomimetic pressure gradient mechanical stimulation system for mimicking cardiac cycle with fluid flow , • Duration: 3 days • 50 mmHg pressure • 10% biaxial stretch • Gradual vs immediate physiological strain , HIPS-CMs on collagen/matrigel coated flexible PDMS membrane , • N/A , • Mimics physiological cardiac cycle • Cell culture chamber performs pumping action to move fluid , • Cells cultured on surface of 3D gels , Rogers et al., .

Techniques: Functional Assay, In Vivo, In Vitro

Journal: Frontiers in Cell and Developmental Biology

Article Title: Naturally Engineered Maturation of Cardiomyocytes

doi: 10.3389/fcell.2017.00050

Figure Lengend Snippet: Stimulation strategies for improving cardiomyocyte maturity .

Article Snippet: , Biomimetic pressure gradient mechanical stimulation system for mimicking cardiac cycle with fluid flow , • Duration: 3 days • 50 mmHg pressure • 10% biaxial stretch • Gradual vs immediate physiological strain , HIPS-CMs on collagen/matrigel coated flexible PDMS membrane , • N/A , • Mimics physiological cardiac cycle • Cell culture chamber performs pumping action to move fluid , • Cells cultured on surface of 3D gels , Rogers et al., .

Techniques: Construct, Cell Culture, Membrane, Expressing, In Vivo, Functional Assay, Permeability, Ubiquitin Proteomics, Control

Journal: Frontiers in Cell and Developmental Biology

Article Title: Naturally Engineered Maturation of Cardiomyocytes

doi: 10.3389/fcell.2017.00050

Figure Lengend Snippet: Bioreactor stimulation systems to improve cardiomyocyte maturity. (A) Cardiac biowire system where PSC-CMs are cultured on a perfusable wire to mimic myocardial fibers. The cardiac biowires are also equipped to be electrically stimulated through two carbon electrodes connected to an electrical stimulation device. Schematic representation of bioreactor from Xiao et al. . (B) A combined mechanical and electrical stimulation bioreactor system. Fibrin hydrogel cardiac constructs are cultured on an inflatable latex tube that can provide cyclic strain to the tissue constructs via a pneumatic system. Exogenous electrical stimulation can also be applied to the constructs by the use of two carbon electrodes connected to an electrical stimulation device. Schematic representation of bioreactor from Morgan and Black . (C) A biomimetic system where PSC-CMs are seeded on to a flexible membrane that can be strained to perform the pumping action required to move fluid. Pressure gradients are used within the input and output to recapitulate each aspect of the cardiac contraction cycle. Schematic representation of bioreactor from Rogers et al. . (D) Large cardiac patch that is cyclic strained by the use of a linear motor that actuates one of the tissue clamped ends. Cannulas are also inserted into the vascular matrix for perfusion of the construct. Schematic representation of bioreactor from Lux et al. .

Article Snippet: , Biomimetic pressure gradient mechanical stimulation system for mimicking cardiac cycle with fluid flow , • Duration: 3 days • 50 mmHg pressure • 10% biaxial stretch • Gradual vs immediate physiological strain , HIPS-CMs on collagen/matrigel coated flexible PDMS membrane , • N/A , • Mimics physiological cardiac cycle • Cell culture chamber performs pumping action to move fluid , • Cells cultured on surface of 3D gels , Rogers et al., .

Techniques: Cell Culture, Construct, Membrane