drg cell line  (Charles River Laboratories)

Journal: The FASEB Journal

Article Title: MRGPRX2 Mediates Mast Cell‐Induced Endometriosis Pain Through the Sensitization of Sensory Neurons via Histamine/ HRH1 / TRPV1 Signaling Pathway

doi: 10.1096/fj.202501493R

Figure Lengend Snippet: Histamine sensitized TRPV1 through HRH1 activation in endometriosis. (A) The expression of histamine receptors, including HRH1, HRH2, HRH3, and HRH4, in F11 cells was determined by PCR using agarose gel electrophoresis. (B) Changes in HRH1 gene expression in F11 cells after treatment with 10 −7 mM histamine for 24 h, as measured by RT–qPCR. (C) Changes in HRH1 protein expression in F11 cells after treatment with 10 −7 mM histamine for 24 h, as detected by western blotting. (D) IHC images showing HRH1 expression in DRG tissue from sham ( n = 8) and EMS mice ( n = 7); 200×, scale bar = 100 μm. The boxed region in the merged images is enlarged in the images on the right (original magnification 400×, bar = 100 μm). (E) IHC scores of the above two groups. (F) Images of IF staining for HRH1 (red), CGRP (green), and cell nuclei (DAPI, blue) in paraffin sections of endometriotic lesions; 400×, scale bar = 100 μm. (G) Fluo‐4 fluorescence intensity of F11 cells from the control, histamine, and histamine+DLT groups. (H) Increase in the Ca 2+ peak (DF/F0) in F11 cells from the control, histamine, and histamine+DLT groups ( n = 3 in each group). Control (B, C, G, and H), F11 cells pretreated with the vehicle control; EMS, mice with endometriosis surgery; His, F11 cells pretreated with histamine (10 −7 mM); histamine, F11 cells pretreated with histamine (10 −7 mM) overnight; histamine + DLT, F11 cells pretreated with histamine (10 −7 mM) and desloratadine (1 μM) overnight; Sham, mice with the sham operation. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, and ns, not significant.

Article Snippet: The DRG cell line (F11) was obtained from the European Collection of Authenticated Cell Cultures (Porton Down, Salisbury, UK).

Techniques: Activation Assay, Expressing, Agarose Gel Electrophoresis, Gene Expression, Quantitative RT-PCR, Western Blot, Staining, Fluorescence, Control

Journal: Cells

Article Title: Integrin-Linked Kinase (ILK) Plays an Important Role in the Laminin-Dependent Development of Dorsal Root Ganglia during Chicken Embryogenesis

doi: 10.3390/cells10071666

Figure Lengend Snippet: ILK is intracellularly localized in DRG neurons and SCPs. ( A – E ) Localization of integrin-linked kinase (ILK) in DRG explant and DRG single cells cultured on the surface coated with poly-D-lysine and additionally laminin-1. DRG explants and cells were cultured for 48 h and then immunostained in the permeabilized conditions with antibodies against ILK, NF-m, Sox10, and ISLET 1/2. To detect F-actin and cell nucleus, the specimens were stained with fluorescently labeled phalloidin and Hoechst 33342, respectively; ( A ) Images representing the whole DRG explant; ( B ) Images showing the presence of ILK in BCM of DRG; ( C ) Images representing ILK’s localization in axon shafts and SCPs. Orange arrows point to the presence of ILK in SCPs, white arrows indicate the localization of ILK in neurites, and the red arrows highlight neurofilament gaps in neurites; ( D ) Images showing ILK’s presence in growth cones and neurites. Yellow arrows point to the gaps in ILK-staining at the sites of neurofilaments accumulations. Enlarged images of these gaps are shown in the insets. White arrowheads highlight the filopodia of growth cones rich in NF-m; ( E ) The images of neuronal cell bodies were obtained from DRG single-cell cultures. White arrows point to axons.

Article Snippet: Therefore, we decided to investigate ILK’s role in the development of Gallus gallus domesticus’s DRG.

Techniques: Cell Culture, Staining, Labeling

Journal: Cells

Article Title: Integrin-Linked Kinase (ILK) Plays an Important Role in the Laminin-Dependent Development of Dorsal Root Ganglia during Chicken Embryogenesis

doi: 10.3390/cells10071666

Figure Lengend Snippet: ILK plays a role in laminin-1-mediated DRG neurite outgrowth. DRG were cultured on a laminin-1-coated surface or with addition to a medium of IKVAV peptide or laminin-1 for 48 h. Separately, control or anti-ILK antibodies were added to the medium. Next, DRG were immunostained against NF-m to detect DRG neurites. ( A ) Representative images of DRG immunostained with anti-NF-m antibodies illustrating the DRG neurite outgrowth; ( B ) Quantification of DRG axonal outgrowth; p ≤ 0.05 (*); p ≤ 0.0001 (****); ( n = 9); ( C ) Representative images of DRG immunostained against anti-NF-m, showing the DRG neurite outgrowth upon alteration of ILK’s function; ( D ) Quantification of DRG axonal outgrowth upon affecting of ILK’s action; p ≤ 0.01 (**); ns—not significant; ( n = 9). Quantitative data ( B , D ) are presented as mean ± SD for ten the longest neurites per each of the nine DRG explants. Individual points represent averaged values obtained from the analysis of each DRG.

Article Snippet: Therefore, we decided to investigate ILK’s role in the development of Gallus gallus domesticus’s DRG.

Techniques: Cell Culture, Control

Journal: Cells

Article Title: Integrin-Linked Kinase (ILK) Plays an Important Role in the Laminin-Dependent Development of Dorsal Root Ganglia during Chicken Embryogenesis

doi: 10.3390/cells10071666

Figure Lengend Snippet: Interruption of ILK’s function results in changed directionality of DRG axons during extension. DRG were cultured on a laminin-1-coated surface, with IKVAV peptide, or with laminin-1 addition to the culture medium for 48 h. Additionally, control or anti-ILK antibodies were added to the medium. Next, DRG were immunostained against NF-m to visualize DRG neurites and analyzed using the OrientationJ plugin of the Fiji application. ( A ) Representative images showing color maps of DRG neurites’ orientation according to the color scale representing the deviation of the direction of neurites from 0° axis. White dashed lines represent the border of BCM; ( B ) Quantitative analysis of neurites’ straightness; ( n = 9). ( C ) Representative images showing color maps of DRG upon treatment with anti-ILK antibodies; ( D ) Quantitative analysis of neurites’ straightness after treatment with anti-ILK antibodies; ( n = 9). Final quantitative data are presented as fold changes of neurites’ straightness (normalized to the control groups for each test. A value higher than 1 indicates a more directed extension of DRG neurites, while values lower than 1 indicate their more chaotic extension. These data were calculated based on AUC from histograms generated by the OrientationJ plugin. More details can be found in the Materials and Methods section and in .

Article Snippet: Therefore, we decided to investigate ILK’s role in the development of Gallus gallus domesticus’s DRG.

Techniques: Cell Culture, Control, Generated

Journal: Cells

Article Title: Integrin-Linked Kinase (ILK) Plays an Important Role in the Laminin-Dependent Development of Dorsal Root Ganglia during Chicken Embryogenesis

doi: 10.3390/cells10071666

Figure Lengend Snippet: ILK does not play a role in laminin-1 mediated migration of Schwann cell precursors. DRG were cultured on a laminin-1-coated surface, with IKVAV peptide, or with laminin-1 for 48 h. Additionally, control or anti-ILK antibodies were added to the medium. Next, DRG were immunostained with anti-Sox10 and anti-NF-m antibodies to detect SCPs and DRG neurites, respectively. ( A ) The scheme of a part of DRG explant showing nuclei of SCPs and their distance of migration, and quantitative analysis of the impact of IKVAV and laminin-1 and disruption of the ILK’s function on the distance traveled by SCPs from the border of BCM; p ≤ 0.001 (***); p ≤ 0.0001 (****); ns—not significant; ( n = 9); ( B ) The scheme of a part of DRG explant showing DRG axons and the length of their halo, and quantitative analysis of the impact of IKVAV and laminin-1 and disruption of the ILK’s function on the length of axonal halo measured from the border of BCM to the tips of the growth cones; p ≤ 0.001 (***); p ≤ 0.0001 (****); ns—not significant; ( n = 9); ( C ) The scheme showing merged A and B schemes. Quantitative analysis of the impact of IKVAV and laminin-1 and disruption of the ILK function on the ratio between distance traveled by SCPs and length of DRG axonal halo; p ≤ 0.01 (**); ns—not significant; ( n = 9). Four sides of each DRG were taken for analysis and then averaged. Individual points represent averaged values obtained from the analysis of each DRG. All bar charts show mean ± SD.

Article Snippet: Therefore, we decided to investigate ILK’s role in the development of Gallus gallus domesticus’s DRG.

Techniques: Migration, Cell Culture, Control, Disruption

Journal: Cells

Article Title: Integrin-Linked Kinase (ILK) Plays an Important Role in the Laminin-Dependent Development of Dorsal Root Ganglia during Chicken Embryogenesis

doi: 10.3390/cells10071666

Figure Lengend Snippet: ILK is important for the number of SCPs migrating from DRG explant when cultured on laminin-1. DRG were cultured on a laminin-1-coated surface or with addition to the medium of IKVAV peptide or laminin-1 for 48 h. Additionally, control or anti-ILK antibodies were added to the medium. The number of SCPs was calculated based on immunostaining against Sox10. ( A ) Representative images showing the nuclei of SCPs present in DRG culture. ( B ) Quantitative analysis of the number of SCPs migrating along axons for DRG; p ≤ 0.0001 (****); ( n = 9); ( C ) Images showing the nuclei of SCPs present in DRG culture after disrupting ILK’s function using anti-ILK antibodies; ( D ) Quantification of the number of SCPs migrating along axons for DRG cultured with antibodies against ILK; p ≤ 0.01 (**); ns—not significant; ( n = 9). All quantitative data are presented as mean ± SD. The number of SCPs was measured for four sides of each analyzed DRG and then averaged. Individual points represent averaged values obtained from the analysis of each DRG.

Article Snippet: Therefore, we decided to investigate ILK’s role in the development of Gallus gallus domesticus’s DRG.

Techniques: Cell Culture, Control, Immunostaining

Journal: Cells

Article Title: Integrin-Linked Kinase (ILK) Plays an Important Role in the Laminin-Dependent Development of Dorsal Root Ganglia during Chicken Embryogenesis

doi: 10.3390/cells10071666

Figure Lengend Snippet: ILK modulates the morphology of a growth cone. DRG were cultured on a laminin-1-coated surface or with addition to the medium of IKVAV peptide or laminin-1 for 48 h. Additionally, control or anti-ILK antibodies were added to the medium. ( A , D ) DRG were then immunostained with NF-m to detect DRG neurites and stained with Alexa Fluor 568-phalloidin to label growth cones’ filopodia rich in F-actin. White arrows point at filopodia rich in F-actin. ( B ) Quantitative analysis of the length of filopodia of DRG growth cones; ns—not significant; ( n = 90). ( C ) Quantitative analysis of the number of filopodia formed per one growth cone; p ≤ 0.0001 (****); ( n = 90). ( E ) Quantification of the length of growth cone filopodia of DRG grown with the addition of anti-ILK antibodies; p ≤ 0.001 (***); ns—not significant; ( n = 90). ( F ) Quantitative analysis of the number of filopodia formed per one growth cone after interruption of ILK’s function; p ≤ 0.001 (***); ns—not significant; ( n = 90). All graphs show mean ± SD. The morphology of ten growth cones per DRG was analyzed.

Article Snippet: Therefore, we decided to investigate ILK’s role in the development of Gallus gallus domesticus’s DRG.

Techniques: Cell Culture, Control, Staining