Journal: PLoS ONE

Article Title: Enhanced β -adrenergic response in mice with dominant-negative expression of the PKD2L1 channel

doi: 10.1371/journal.pone.0261668

Figure Lengend Snippet: A) Molecular characterization of PKD2L1del overexpressed in Tg mice. Schematic of the CMV promoter/PKD2L1del construct used for the generation of PKD2L1del-Tg mice. The deletion site is also shown. The primer sets for each PCR amplification (DNA typing and RT-PCR) are indicated with arrows. The inset shows predicted schematic transmembrane constructs of full-length PKD2L1 (upper panel) and PKD2L1del. Scale bar = 1 kb. B) DNA typing of Tg mice. Tg-A mice contained a single copy of the external gene, whereas mice in Tg-B contained two copies. RT-PCR analysis of cardiac tissue from Tg mice. Tg-B mice showed enhanced PKD2L1 expression in the heart. NC , negative control without cDNA. The names of the Tg lines (A and B) are indicated. C) Western blot analysis of the hearts from the WT and Tg mice. Representative immunoblots of membranes from WT and Tg mice analyzed for the expression of PKD2L1 (left panel) and GAPDH (right panel).

Article Snippet: Commercially available polyclonal antibodies specific for PKD2L1 (Novus Biologicals, Centennial, CO, USA).

Techniques: Construct, Amplification, Reverse Transcription Polymerase Chain Reaction, Expressing, Negative Control, Western Blot

Journal: PLoS ONE

Article Title: Enhanced β -adrenergic response in mice with dominant-negative expression of the PKD2L1 channel

doi: 10.1371/journal.pone.0261668

Figure Lengend Snippet: A) Typical RT-PCR data from kidney and heart tissues collected from WT and Tg mice. Amplified sequences are shown. Increased expression levels of PKD2L1 were confirmed in kidney and heart tissues from Tg mice. The expression levels of PKD1, PKD2, and PKD2L2 were assessed. Expression of β-actin was evaluated as a control. The primer sets used for PCR amplification are shown. B) Representative ECG tracings of WT and Tg mice under basal conditions (i). Statistical analysis of the heart rates of WT and Tg mice (ii). There were no significant differences in heart rate between WT ( n = 10) and Tg ( n = 6) mice under basal conditions. Isoproterenol administration resulted in increased heart rate changes in Tg mice (iii). * P < 0.05, between WT (n = 10) and Tg (n = 6) mice. Error bars indicate the standard error of the mean.

Article Snippet: Commercially available polyclonal antibodies specific for PKD2L1 (Novus Biologicals, Centennial, CO, USA).

Techniques: Reverse Transcription Polymerase Chain Reaction, Amplification, Expressing, Control

Journal: Cellular signalling

Article Title: Vegfr3 activation of Pkd2l1 + CSF-cNs triggers the neural stem cell response in spinal cord injury.

doi: 10.1016/j.cellsig.2025.111675

Figure Lengend Snippet: Fig. 1. Vegfr3 is expressed primarily in Pkd2l1+ CSF-cNs. (A) A diagram illustrating the process of isolating and culturing primary CSF-cNs. (B) Pkd2l1+ CSF-cNs expressed Vegfr1, Vegfr2, and Vegfr3, with Vegfr3 showing the highest expression in vitro. (C) Western blot analysis of Vegfr1, Vegfr2, and Vegfr3 in Pkd2l1+ CSF- cNs. (D) Strategy diagram for the construction of Pkd2l1-2 A-CreERT2 × R26-CAG-LSL-ZsGreen transgenic mice, outlining the genetic modifications and breeding strategy employed. (E) Immunofluorescence colabeling of Pkd2l1 and the inherent ZsGreen fluorescence in Pkd2l1-2 A-CreERT2 × R26-CAG-LSL-ZsGreen transgenic mice. The overlap of the Pkd2l1 and ZsGreen signals confirmed the successful generation of the transgenic model. (F) Coexpression analysis showing Vegfr3 localization in Pkd2l1+ CSF-cNs. Double immunofluorescence staining revealed the colocalization of Vegfr3 with Pkd2l1+ cells, indicating that Vegfr3 is expressed in these neurons. (G) Immunofluorescence images illustrate the lack of colocalization between Vegfr3 and Foxj1, a specific marker for ependymal cells, in both the sham and SCI 7 d groups. These findings indicate that Vegfr3+ cells are distinct from ependymal cells. (H) Immunofluorescence images show the absence of colocalization between Vegfr3 and ALDH1L1, an astrocyte marker, in both the sham and SCI 7 d groups. Scale bar: 50 μm (B, E, F, G, H).

Article Snippet: Primary antibodies targeting the following proteins were used: Pkd2l1 (#AB9084, 1:700, Merck Millipore), CD133 (#66666–1-Ig, 1:500, Proteintech), GFAP (#3670, 1:300, Cell Signaling Technology), EGFR (#66863–1-lg, 1:500, Proteintech), and Nestin (1:300, Sc-6251, Santa Cruz Biotechnology).

Techniques: Expressing, In Vitro, Western Blot, Transgenic Assay, Immunofluorescence, Fluorescence, Double Immunofluorescence Staining, Marker

Journal: Cellular signalling

Article Title: Vegfr3 activation of Pkd2l1 + CSF-cNs triggers the neural stem cell response in spinal cord injury.

doi: 10.1016/j.cellsig.2025.111675

Figure Lengend Snippet: Fig. 3. Role of Vegfr3 in the proliferation and activation of Pkd2l1+ CSF-cNs. (A, B) The overexpression and knockdown efficiencies of Vegfr3 in Pkd2l1+ CSF-cNs were confirmed through qRT-PCR and Western blotting following transfection. (C) Representative images of CSF-cNs and the average diameter of neurospheres with or without Vegfr3 overexpression or inhibition. (D) Representative immu nofluorescence images and quantification of EdU+ Pkd2l1+ CSF-cNs with or without Vegfr3 overexpression or inhibition. (E) Representative immunofluorescence images and quantification of pHH3+ Pkd2l1+ CSF-cNs with or without Vegfr3 overexpression or inhibition. (F) A CCK-8 assay was used to assess the viability of Pkd2l1+ CSF-cNs with either Vegfr3 overexpression or knockdown. (G) Representative Western blots showing the protein levels of EGFR, Ascl, and Sox2 in Pkd2l1+

Article Snippet: Primary antibodies targeting the following proteins were used: Pkd2l1 (#AB9084, 1:700, Merck Millipore), CD133 (#66666–1-Ig, 1:500, Proteintech), GFAP (#3670, 1:300, Cell Signaling Technology), EGFR (#66863–1-lg, 1:500, Proteintech), and Nestin (1:300, Sc-6251, Santa Cruz Biotechnology).

Techniques: Activation Assay, Over Expression, Knockdown, Quantitative RT-PCR, Western Blot, Transfection, Inhibition, Immunofluorescence, CCK-8 Assay

Journal: Cell death discovery

Article Title: Pkd2l1 deletion inhibits the neurogenesis of cerebrospinal fluid-contacting neurons and impedes spinal cord injury repair.

doi: 10.1038/s41420-025-02492-y

Figure Lengend Snippet: Fig. 1 Pkd2l1+ CSF-cNs have the characteristics of both immature neurons and adult NSCs. a Immunofluorescence staining showing expression of the NSC marker Nestin in Pkd2l1+ CSF-cNs. b Pkd2l1+ CSF-cNs expressed the NSC marker Sox2 by immunofluorescence staining. c Pkd2l1+ CSF-cNs expressed the NSC marker CD133 by immunofluorescence staining. d Pkd2l1+ CSF-cNs did not express the NSC marker GFAP by immunofluorescence staining. e Pkd2l1+ CSF-cNs expressed the neuroblast marker doublecortin (Dcx) by immunofluorescence staining. f Immunofluorescence staining showed that Pkd2l1+ CSF-cNs expressed the immature neuronal marker βIII-tubulin (red). Scale bars, 20 µm. Nuclear counterstaining, DAPI.

Article Snippet: Pkd2l1 WT mice were acquired from Beijing Viton Lihua Laboratory Animal Technology Co., Ltd. Pkd2l1 knockout mice were purchased from Cyagen Biosciences (Guangzhou, China).

Techniques: Staining, Expressing, Marker

Journal: Cell death discovery

Article Title: Pkd2l1 deletion inhibits the neurogenesis of cerebrospinal fluid-contacting neurons and impedes spinal cord injury repair.

doi: 10.1038/s41420-025-02492-y

Figure Lengend Snippet: Fig. 2 Pkd2l1 deletion results in the loss of CSF-cNs. a Schematic showing the generation and identification of genetically homogeneous Pkd2l1−/−mice. b, c Immunofluorescence staining of spinal cord sections showed that Pkd2l1 deletion caused the obviously reduced proliferation of GABA-positive cells after SCI (GABA: CSF-cNs marker) (n = 6). d, e The depletion of Pkd2l1 caused a significant reduction in Gata2-positive cells after SCI, as shown by immunofluorescence staining (Gata2: CSF-cNs marker) (n = 6). f, g Immunofluorescence staining shows that Pkd2l1 deletion resulted in almost the complete absence of Gata3-positive cell proliferation after SCI (Gata3: CSF-cNs marker) (n = 6). ***p < 0.001. Data are presented as the mean ± SD. Scale bars, 50 µm. Nuclear counterstaining, DAPI.

Article Snippet: Pkd2l1 WT mice were acquired from Beijing Viton Lihua Laboratory Animal Technology Co., Ltd. Pkd2l1 knockout mice were purchased from Cyagen Biosciences (Guangzhou, China).

Techniques: Staining, Marker

Journal: Cell death discovery

Article Title: Pkd2l1 deletion inhibits the neurogenesis of cerebrospinal fluid-contacting neurons and impedes spinal cord injury repair.

doi: 10.1038/s41420-025-02492-y

Figure Lengend Snippet: Fig. 3 Pkd2l1 deletion suppresses the proliferation of CSF-cNs and leads to the loss of migratory CSF-cNs following SCI. a, b The EdU proliferation assay revealed a significant reduction in EdU-positive cells in Pkd2l1−/−mice compared to wild-type counterparts (n = 6). c, d Immunofluorescence analysis revealed a notable decrease in Dcx-positive cells following SCI in Pkd2l1−/−mice compared to their wild- type counterparts (n = 6). *p < 0.05, **p < 0.01. Data are presented as the mean ± SD. Scale bars, 50 µm.

Article Snippet: Pkd2l1 WT mice were acquired from Beijing Viton Lihua Laboratory Animal Technology Co., Ltd. Pkd2l1 knockout mice were purchased from Cyagen Biosciences (Guangzhou, China).

Techniques: Proliferation Assay

Journal: Cell death discovery

Article Title: Pkd2l1 deletion inhibits the neurogenesis of cerebrospinal fluid-contacting neurons and impedes spinal cord injury repair.

doi: 10.1038/s41420-025-02492-y

Figure Lengend Snippet: Fig. 4 Pkd2l1 deletion retards the self-renewal of NSCs surrounding the central canal of the spinal cord after SCI. a, b Immunofluorescence staining showed that Pkd2l1−/−mice had almost no Nestin-positive cells surrounding the central canal of the spinal cord after SCI (n = 6). c, d Sox2 expression evidently decreased in Pkd2l1−/−mice after SCI, as shown by immunofluorescence staining (n = 6). e, f GFAP expression was reduced in Pkd2l1−/−mice after SCI, as shown by immunofluorescence staining (n = 6). ***p < 0.001. Data are presented as the mean ± SD. Scale bars, 50 µm. Nuclear counterstaining, DAPI.

Article Snippet: Pkd2l1 WT mice were acquired from Beijing Viton Lihua Laboratory Animal Technology Co., Ltd. Pkd2l1 knockout mice were purchased from Cyagen Biosciences (Guangzhou, China).

Techniques: Staining, Expressing

Journal: Cell death discovery

Article Title: Pkd2l1 deletion inhibits the neurogenesis of cerebrospinal fluid-contacting neurons and impedes spinal cord injury repair.

doi: 10.1038/s41420-025-02492-y

Figure Lengend Snippet: Fig. 5 Pkd2l1 deletion retards myelin and neuronal regeneration following SCI. a, b Luxol Fast Blue staining showed that myelin regeneration was almost depleted in Pkd2l1−/−mice (n = 6). c, d Nissl staining showed that Pkd2l1−/−mice had a notable reduction in Nissl bodies surrounding the central canal following SCI (n = 6). *p < 0.05, **p < 0.01, ***p < 0.001. Data are presented as the mean ± SD: scale bars, 100 µm.

Article Snippet: Pkd2l1 WT mice were acquired from Beijing Viton Lihua Laboratory Animal Technology Co., Ltd. Pkd2l1 knockout mice were purchased from Cyagen Biosciences (Guangzhou, China).

Techniques: Staining

Journal: Cell death discovery

Article Title: Pkd2l1 deletion inhibits the neurogenesis of cerebrospinal fluid-contacting neurons and impedes spinal cord injury repair.

doi: 10.1038/s41420-025-02492-y

Figure Lengend Snippet: Fig. 7 Pkd2l1−/−mice restrained motor function recovery following SCI. a Measurement of BMS scores showed that the knockout of Pkd2l1 restricted motor function recovery in mice after SCI. b The loss of motor function in Pkd2l1−/−mice was evaluated based on the status of the hind limbs on day 35 following SCI (n = 6). *p < 0.05. compared to the Pkd2l1−/−SCI group. Data are presented as the mean ± SD.

Article Snippet: Pkd2l1 WT mice were acquired from Beijing Viton Lihua Laboratory Animal Technology Co., Ltd. Pkd2l1 knockout mice were purchased from Cyagen Biosciences (Guangzhou, China).

Techniques: Knock-Out

Journal: Cell death discovery

Article Title: Pkd2l1 deletion inhibits the neurogenesis of cerebrospinal fluid-contacting neurons and impedes spinal cord injury repair.

doi: 10.1038/s41420-025-02492-y

Figure Lengend Snippet: Fig. 6 Pkd2l1 deletion promotes pericentral canal vacuolar degeneration and glial scar formation following SCI. a H&E staining showed that Pkd2l1−/−mice had more pericentral canal vacuolar degeneration after SCI. b, c Masson staining conducted on the transverse section of spinal cord tissue at the thoracic 10 (T10) level revealed that Pkd2l1−/−mice exhibited pronounced accumulation of collagen fibers, leading to the formation of glial scars around the central canal following SCI (n = 6). *p < 0.05. Data are presented as the mean ± SD. Scale bars, 100 µm.

Article Snippet: Pkd2l1 WT mice were acquired from Beijing Viton Lihua Laboratory Animal Technology Co., Ltd. Pkd2l1 knockout mice were purchased from Cyagen Biosciences (Guangzhou, China).

Techniques: Staining

Journal: PLoS ONE

Article Title: Expression of Genes Encoding Multi-Transmembrane Proteins in Specific Primate Taste Cell Populations

doi: 10.1371/journal.pone.0007682

Figure Lengend Snippet: A–F, Double label in situ hybridization (ISH) for TRPM5 and PKD1L3. TRPM5 (A, D) and PKD1L3 (B, E) are expressed in different cells in the merged images (C, F). G–L, Double label ISH for PKD2L1 and PKD1L3. PKD2L1 (G, J) and PKD1L3 (H, K) are expressed in similar cells in the merged images (I, L). Identical results were obtained using double label fluorescent ISH (A–C and G–I) or double label colorimetric-fluorescent ISH (D–F and J–L). Double label fluorescent ISH images show multiple taste buds whereas double label colorimetric-fluorescent ISH images show a single taste bud. Scale bar is 40µm in A and represents scale for A–C and G–I, 20µm in F and represents scale for D–F and J–L. Images are from primate CV papilla. M, Pie chart illustrating fraction of cells expressing TRPM5, PKD1L3, or both TRPM5 and PKD1L3. N, Pie chart illustrating fraction of cells expressing PKD2L1, PKD1L3, or both PKD2L1 and PKD1L3.

Article Snippet: Semi-quantitative PCR was performed using the Mx3000P® QPCR System (Stratagene, La Jolla CA) using TaqMan Universal PCR Master Mix (Applied Biosystems, Foster City, CA), 10 ng of cDNA, and the following inventoried TaqMan assays: TRPM5 (Hs00175822_m1), PKD2L1 (Hs00175850_m1), ANO7 (Hs00417639_m1), CALHM1 (Hs00736332_m1), CALHM3 (Hs00699323_m1), MCTP1 (Hs00226801_m1), SV2B (Hs00208178_m1), TMEM44 (Hs00961693_m1), and GAPDH (Hs99999905_m1).

Techniques: In Situ Hybridization, Expressing

Journal: PLoS ONE

Article Title: Expression of Genes Encoding Multi-Transmembrane Proteins in Specific Primate Taste Cell Populations

doi: 10.1371/journal.pone.0007682

Figure Lengend Snippet: Double label in situ hybridization (ISH) for MCTP1 and TRPM5 (A–F). MCTP1 (A, D) and TRPM5 (B, E) are expressed in similar cells in the merged images (C, F). Double label ISH for MCTP1 and PKD2L1 (G–L). MCTP1 (G, J) and PKD2L1 (H, K) are expressed in different cells in the merged images (I, L). Images in D–F and J–L depict single taste buds at higher magnification. Scale bar is 40µm in C and represents scale for A–C and G–I. Scale bar is 25µm in F and represents scale for D–F and J–L. Images are from mouse CV papilla. M, Pie chart illustrating fraction of cells expressing MCTP1, TRPM5, or both MCTP1 and TRPM5. N, Pie chart illustrating fraction of cells expressing MCTP1, PKD2L1, or both MCTP1 and PKD2L1.

Article Snippet: Semi-quantitative PCR was performed using the Mx3000P® QPCR System (Stratagene, La Jolla CA) using TaqMan Universal PCR Master Mix (Applied Biosystems, Foster City, CA), 10 ng of cDNA, and the following inventoried TaqMan assays: TRPM5 (Hs00175822_m1), PKD2L1 (Hs00175850_m1), ANO7 (Hs00417639_m1), CALHM1 (Hs00736332_m1), CALHM3 (Hs00699323_m1), MCTP1 (Hs00226801_m1), SV2B (Hs00208178_m1), TMEM44 (Hs00961693_m1), and GAPDH (Hs99999905_m1).

Techniques: In Situ Hybridization, Expressing

Journal: PLoS ONE

Article Title: Expression of Genes Encoding Multi-Transmembrane Proteins in Specific Primate Taste Cell Populations

doi: 10.1371/journal.pone.0007682

Figure Lengend Snippet: Double label in situ hybridization (ISH) for ANO7 and TRPM5 (A–F). ANO7 (A, D) and TRPM5 (B, E) are expressed in similar cells in the merged images (C, F). Double label ISH for ANO7 and PKD2L1 (G–L). ANO7 (G, J) and PKD2L1 (H, K) are expressed in different cells in the merged images (I, L). Images in D–F and J–L depict single taste buds at higher magnification. Scale bar is 40µm in C and represents scale for A–C and G–I. Scale bar is 10µm in F and represents scale for D–F and J–L. Images are from mouse CV papilla. M, Pie chart illustrating fraction of cells expressing ANO7, TRPM5, or both ANO7 and TRPM5. N, Pie chart illustrating fraction of cells expressing ANO7, PKD2L1, or both ANO7 and PKD2L1.

Article Snippet: Semi-quantitative PCR was performed using the Mx3000P® QPCR System (Stratagene, La Jolla CA) using TaqMan Universal PCR Master Mix (Applied Biosystems, Foster City, CA), 10 ng of cDNA, and the following inventoried TaqMan assays: TRPM5 (Hs00175822_m1), PKD2L1 (Hs00175850_m1), ANO7 (Hs00417639_m1), CALHM1 (Hs00736332_m1), CALHM3 (Hs00699323_m1), MCTP1 (Hs00226801_m1), SV2B (Hs00208178_m1), TMEM44 (Hs00961693_m1), and GAPDH (Hs99999905_m1).

Techniques: In Situ Hybridization, Expressing

Journal: PLoS ONE

Article Title: Expression of Genes Encoding Multi-Transmembrane Proteins in Specific Primate Taste Cell Populations

doi: 10.1371/journal.pone.0007682

Figure Lengend Snippet: Section of human CV papilla before (A) and after (B) laser capture microdissection of taste buds. Collected taste bud regions (C), were isolated from CV papilla and used for molecular analysis of gene expression. A laser beam was used to cut the perimeter of taste buds and physically separate them from surrounding lingual epithelium. Taste buds were next lifted away from the tissue section with an adhesive cap. Panel C is an image of six isolated taste bud regions, devoid of surrounding lingual epithelium and connective tissue, on the adhesive cap. Scale bar is 40µm. Semi-quantitative PCR (D) for known taste genes (TRPM5 and PKD2L1), genes predicted or known to encode transmembrane proteins, and the housekeeping gene GAPDH in isolated CV taste buds (black bars) or non-gustatory lingual epithelium (white bars) collected by laser capture microdissection. Relative expression is shown on a logarithmic scale.

Article Snippet: Semi-quantitative PCR was performed using the Mx3000P® QPCR System (Stratagene, La Jolla CA) using TaqMan Universal PCR Master Mix (Applied Biosystems, Foster City, CA), 10 ng of cDNA, and the following inventoried TaqMan assays: TRPM5 (Hs00175822_m1), PKD2L1 (Hs00175850_m1), ANO7 (Hs00417639_m1), CALHM1 (Hs00736332_m1), CALHM3 (Hs00699323_m1), MCTP1 (Hs00226801_m1), SV2B (Hs00208178_m1), TMEM44 (Hs00961693_m1), and GAPDH (Hs99999905_m1).

Techniques: Laser Capture Microdissection, Isolation, Gene Expression, Adhesive, Real-time Polymerase Chain Reaction, Expressing