trpv5  (Alomone Labs)


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    Alomone Labs trpv5
    Microscopic images showing localization of <t>TRPV5</t> in duck sperm. (A-B) Confocal microscopic images depicting the localization of TRPV5 (green) as detected by two different antibodies and Nucleus (blue) by DAPI. Mitochondria (red) is labelled by Mitotracker Red dye in A and C to highlight the channel expression in the mitochondrial region. ( C ) SR-SIM images of TRPV5 localization (using Ab1 antibody) at the head (left) and tail (right) of duck sperm is shown. ( D ) Zoomed up image of neck region of sperm depicting the absence of TRPV5 (green) in the neck region. The head (blue) and arrows mark the start and end point of mitochondrial region.
    Trpv5, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/trpv5/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    trpv5 - by Bioz Stars, 2021-12
    93/100 stars

    Images

    1) Product Images from "Differential expression and localization of thermosensitive Transient Receptor Potential Vanilloid (TRPV) channels in the mature sperm of white pekin duck (Anas platyrhynchos)"

    Article Title: Differential expression and localization of thermosensitive Transient Receptor Potential Vanilloid (TRPV) channels in the mature sperm of white pekin duck (Anas platyrhynchos)

    Journal: bioRxiv

    doi: 10.1101/2020.02.10.941732

    Microscopic images showing localization of TRPV5 in duck sperm. (A-B) Confocal microscopic images depicting the localization of TRPV5 (green) as detected by two different antibodies and Nucleus (blue) by DAPI. Mitochondria (red) is labelled by Mitotracker Red dye in A and C to highlight the channel expression in the mitochondrial region. ( C ) SR-SIM images of TRPV5 localization (using Ab1 antibody) at the head (left) and tail (right) of duck sperm is shown. ( D ) Zoomed up image of neck region of sperm depicting the absence of TRPV5 (green) in the neck region. The head (blue) and arrows mark the start and end point of mitochondrial region.
    Figure Legend Snippet: Microscopic images showing localization of TRPV5 in duck sperm. (A-B) Confocal microscopic images depicting the localization of TRPV5 (green) as detected by two different antibodies and Nucleus (blue) by DAPI. Mitochondria (red) is labelled by Mitotracker Red dye in A and C to highlight the channel expression in the mitochondrial region. ( C ) SR-SIM images of TRPV5 localization (using Ab1 antibody) at the head (left) and tail (right) of duck sperm is shown. ( D ) Zoomed up image of neck region of sperm depicting the absence of TRPV5 (green) in the neck region. The head (blue) and arrows mark the start and end point of mitochondrial region.

    Techniques Used: Expressing

    Microscopic images showing localization of TRPV6 in duck sperm. (A-B) Confocal microscopic images depicting the localization of TRPV6 (green) as detected by two different antibodies and Nucleus (blue) by DAPI. Mitochondria (red) is labelled by Mitotracker Red dye in A and C to highlight the channel expression in the mitochondrial region. ( C ) SR-SIM images of TRPV6 localization (using Ab1 antibody) at the head (left) and tail (right) of duck sperm is shown. ( D ) Zoomed up image of neck region of sperm depicting the absence of TRPV6 (green) in the neck region. The head (blue) and arrows mark the start and end point of mitochondrial region.
    Figure Legend Snippet: Microscopic images showing localization of TRPV6 in duck sperm. (A-B) Confocal microscopic images depicting the localization of TRPV6 (green) as detected by two different antibodies and Nucleus (blue) by DAPI. Mitochondria (red) is labelled by Mitotracker Red dye in A and C to highlight the channel expression in the mitochondrial region. ( C ) SR-SIM images of TRPV6 localization (using Ab1 antibody) at the head (left) and tail (right) of duck sperm is shown. ( D ) Zoomed up image of neck region of sperm depicting the absence of TRPV6 (green) in the neck region. The head (blue) and arrows mark the start and end point of mitochondrial region.

    Techniques Used: Expressing

    Endogenous expression of TRPV channels in duck sperm. Western blot analysis of duck sperm extracts probed with different TRPV-specific antibodies are shown. A. TRPV1 specific band is detected by a specific antibody (directed against the C-terminus of TRPV1, Alomone Labs) in absence but not in presence of its blocking peptide; B. Western blot analysis with antibody that detects TRPV2 (raised against the C-terminus, Alomone Labs). C. Two different antibodies detecting TRPV3 [raised against the C-terminus, (Ab1: Alomone Labs) and N-terminus (Ab3: Sigma Aldrich)] detect similar expression pattern of TRPV3. D. Two different antibodies raised against the TRPV4 [raised against C-terminus, Ab1: Alomone Labs) and N-terminus (Ab3: Sigma Aldrich)] detect TRPV4 at the expected size. E. Two different antibodies raised against the C-terminus of TRPV5 (Ab1: Alomone Labs and Ab2: Sigma-Aldrich) detects TRPV5 at expected size. F. A specific antibody raised against the C-terminus of TRPV6 (Ab-1: Alomone Labs) detects TRPV6 in absence but not in presence of its blocking peptide.
    Figure Legend Snippet: Endogenous expression of TRPV channels in duck sperm. Western blot analysis of duck sperm extracts probed with different TRPV-specific antibodies are shown. A. TRPV1 specific band is detected by a specific antibody (directed against the C-terminus of TRPV1, Alomone Labs) in absence but not in presence of its blocking peptide; B. Western blot analysis with antibody that detects TRPV2 (raised against the C-terminus, Alomone Labs). C. Two different antibodies detecting TRPV3 [raised against the C-terminus, (Ab1: Alomone Labs) and N-terminus (Ab3: Sigma Aldrich)] detect similar expression pattern of TRPV3. D. Two different antibodies raised against the TRPV4 [raised against C-terminus, Ab1: Alomone Labs) and N-terminus (Ab3: Sigma Aldrich)] detect TRPV4 at the expected size. E. Two different antibodies raised against the C-terminus of TRPV5 (Ab1: Alomone Labs and Ab2: Sigma-Aldrich) detects TRPV5 at expected size. F. A specific antibody raised against the C-terminus of TRPV6 (Ab-1: Alomone Labs) detects TRPV6 in absence but not in presence of its blocking peptide.

    Techniques Used: Expressing, Western Blot, Blocking Assay

    2) Product Images from "Differential expression and localization of thermosensitive Transient Receptor Potential Vanilloid (TRPV) channels in the mature sperm of white pekin duck (Anas platyrhynchos)"

    Article Title: Differential expression and localization of thermosensitive Transient Receptor Potential Vanilloid (TRPV) channels in the mature sperm of white pekin duck (Anas platyrhynchos)

    Journal: bioRxiv

    doi: 10.1101/2020.02.10.941732

    Microscopic images showing localization of TRPV5 in duck sperm. (A-B) Confocal microscopic images depicting the localization of TRPV5 (green) as detected by two different antibodies and Nucleus (blue) by DAPI. Mitochondria (red) is labelled by Mitotracker Red dye in A and C to highlight the channel expression in the mitochondrial region. ( C ) SR-SIM images of TRPV5 localization (using Ab1 antibody) at the head (left) and tail (right) of duck sperm is shown. ( D ) Zoomed up image of neck region of sperm depicting the absence of TRPV5 (green) in the neck region. The head (blue) and arrows mark the start and end point of mitochondrial region.
    Figure Legend Snippet: Microscopic images showing localization of TRPV5 in duck sperm. (A-B) Confocal microscopic images depicting the localization of TRPV5 (green) as detected by two different antibodies and Nucleus (blue) by DAPI. Mitochondria (red) is labelled by Mitotracker Red dye in A and C to highlight the channel expression in the mitochondrial region. ( C ) SR-SIM images of TRPV5 localization (using Ab1 antibody) at the head (left) and tail (right) of duck sperm is shown. ( D ) Zoomed up image of neck region of sperm depicting the absence of TRPV5 (green) in the neck region. The head (blue) and arrows mark the start and end point of mitochondrial region.

    Techniques Used: Expressing

    Microscopic images showing localization of TRPV6 in duck sperm. (A-B) Confocal microscopic images depicting the localization of TRPV6 (green) as detected by two different antibodies and Nucleus (blue) by DAPI. Mitochondria (red) is labelled by Mitotracker Red dye in A and C to highlight the channel expression in the mitochondrial region. ( C ) SR-SIM images of TRPV6 localization (using Ab1 antibody) at the head (left) and tail (right) of duck sperm is shown. ( D ) Zoomed up image of neck region of sperm depicting the absence of TRPV6 (green) in the neck region. The head (blue) and arrows mark the start and end point of mitochondrial region.
    Figure Legend Snippet: Microscopic images showing localization of TRPV6 in duck sperm. (A-B) Confocal microscopic images depicting the localization of TRPV6 (green) as detected by two different antibodies and Nucleus (blue) by DAPI. Mitochondria (red) is labelled by Mitotracker Red dye in A and C to highlight the channel expression in the mitochondrial region. ( C ) SR-SIM images of TRPV6 localization (using Ab1 antibody) at the head (left) and tail (right) of duck sperm is shown. ( D ) Zoomed up image of neck region of sperm depicting the absence of TRPV6 (green) in the neck region. The head (blue) and arrows mark the start and end point of mitochondrial region.

    Techniques Used: Expressing

    Endogenous expression of TRPV channels in duck sperm. Western blot analysis of duck sperm extracts probed with different TRPV-specific antibodies are shown. A. TRPV1 specific band is detected by a specific antibody (directed against the C-terminus of TRPV1, Alomone Labs) in absence but not in presence of its blocking peptide; B. Western blot analysis with antibody that detects TRPV2 (raised against the C-terminus, Alomone Labs). C. Two different antibodies detecting TRPV3 [raised against the C-terminus, (Ab1: Alomone Labs) and N-terminus (Ab3: Sigma Aldrich)] detect similar expression pattern of TRPV3. D. Two different antibodies raised against the TRPV4 [raised against C-terminus, Ab1: Alomone Labs) and N-terminus (Ab3: Sigma Aldrich)] detect TRPV4 at the expected size. E. Two different antibodies raised against the C-terminus of TRPV5 (Ab1: Alomone Labs and Ab2: Sigma-Aldrich) detects TRPV5 at expected size. F. A specific antibody raised against the C-terminus of TRPV6 (Ab-1: Alomone Labs) detects TRPV6 in absence but not in presence of its blocking peptide.
    Figure Legend Snippet: Endogenous expression of TRPV channels in duck sperm. Western blot analysis of duck sperm extracts probed with different TRPV-specific antibodies are shown. A. TRPV1 specific band is detected by a specific antibody (directed against the C-terminus of TRPV1, Alomone Labs) in absence but not in presence of its blocking peptide; B. Western blot analysis with antibody that detects TRPV2 (raised against the C-terminus, Alomone Labs). C. Two different antibodies detecting TRPV3 [raised against the C-terminus, (Ab1: Alomone Labs) and N-terminus (Ab3: Sigma Aldrich)] detect similar expression pattern of TRPV3. D. Two different antibodies raised against the TRPV4 [raised against C-terminus, Ab1: Alomone Labs) and N-terminus (Ab3: Sigma Aldrich)] detect TRPV4 at the expected size. E. Two different antibodies raised against the C-terminus of TRPV5 (Ab1: Alomone Labs and Ab2: Sigma-Aldrich) detects TRPV5 at expected size. F. A specific antibody raised against the C-terminus of TRPV6 (Ab-1: Alomone Labs) detects TRPV6 in absence but not in presence of its blocking peptide.

    Techniques Used: Expressing, Western Blot, Blocking Assay

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    Alomone Labs rabbit anti trpv5
    Channel characteristics of wild-type and mutant <t>TRPV5.</t> ( A ) Whole-cell currents in TRPV5-WT (V5-WT) and TRPV5-682P (V5-S682P) injected Xenopus oocytes recorded in response to 300 ms test pulses to various potentials (from −100 to +60 mV in 10 mV increments). Holding potential, 0 mV (N = 5). ( B ) Mean current-voltage relationships for TRPV5-WT and TRPV5-682P channels (N = 5). These current-voltage relationships are similar to those reported for TRPV5 channels [62] . ( C ) Mean whole-cell tail currents measured in TRPV5-WT and TRPV5-682P injected Xenopus oocytes during test potentials applied in 10 mV increments from −70 to +40 mV after a pre-pulse to −100 mV in TRPV5-WT and TRPV5-682P channels (N = 5). ( D ) Time-dependent inhibition of TRPV-WT and TRPV5-682P whole-cell currents. Oocytes were stimulated every 1 s. The peak current amplitude was normalised to that recorded during the first pulse (N = 4). ( E ) Representative trace of Fura-2 ratio in HEK293 cells transiently transfected with an empty EGFP vector (mock), or EGFP-tagged TRPV5-WT or TRPV5-S682P. Cells expressing EGFP were selected and monitored for changes in intracellular Ca 2+ levels when extracellular Ca 2+ concentrations were varied from 1.4 mM Ca 2+ to 0 mM Ca 2+ (2 mM EDTA) and 1.4 mM Ca 2+ which was facilitated by superfusion. ( F ) Fura-2 levels under resting conditions (t0), minimal Fura-2 ratio after EDTA treatment (tmin) and peak level (tmax) upon administration of 1.4 mM Ca 2+ after EDTA treatment. Average data of cells transfected with the empty vector (n = 7), TRPV5-wt (n = 24) and TRPV5-S682P (N = 24) from at least three independent experiments. * p
    Rabbit Anti Trpv5, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti trpv5/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti trpv5 - by Bioz Stars, 2021-12
    93/100 stars
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    Channel characteristics of wild-type and mutant TRPV5. ( A ) Whole-cell currents in TRPV5-WT (V5-WT) and TRPV5-682P (V5-S682P) injected Xenopus oocytes recorded in response to 300 ms test pulses to various potentials (from −100 to +60 mV in 10 mV increments). Holding potential, 0 mV (N = 5). ( B ) Mean current-voltage relationships for TRPV5-WT and TRPV5-682P channels (N = 5). These current-voltage relationships are similar to those reported for TRPV5 channels [62] . ( C ) Mean whole-cell tail currents measured in TRPV5-WT and TRPV5-682P injected Xenopus oocytes during test potentials applied in 10 mV increments from −70 to +40 mV after a pre-pulse to −100 mV in TRPV5-WT and TRPV5-682P channels (N = 5). ( D ) Time-dependent inhibition of TRPV-WT and TRPV5-682P whole-cell currents. Oocytes were stimulated every 1 s. The peak current amplitude was normalised to that recorded during the first pulse (N = 4). ( E ) Representative trace of Fura-2 ratio in HEK293 cells transiently transfected with an empty EGFP vector (mock), or EGFP-tagged TRPV5-WT or TRPV5-S682P. Cells expressing EGFP were selected and monitored for changes in intracellular Ca 2+ levels when extracellular Ca 2+ concentrations were varied from 1.4 mM Ca 2+ to 0 mM Ca 2+ (2 mM EDTA) and 1.4 mM Ca 2+ which was facilitated by superfusion. ( F ) Fura-2 levels under resting conditions (t0), minimal Fura-2 ratio after EDTA treatment (tmin) and peak level (tmax) upon administration of 1.4 mM Ca 2+ after EDTA treatment. Average data of cells transfected with the empty vector (n = 7), TRPV5-wt (n = 24) and TRPV5-S682P (N = 24) from at least three independent experiments. * p

    Journal: PLoS ONE

    Article Title: Autosomal Dominant Hypercalciuria in a Mouse Model Due to a Mutation of the Epithelial Calcium Channel, TRPV5

    doi: 10.1371/journal.pone.0055412

    Figure Lengend Snippet: Channel characteristics of wild-type and mutant TRPV5. ( A ) Whole-cell currents in TRPV5-WT (V5-WT) and TRPV5-682P (V5-S682P) injected Xenopus oocytes recorded in response to 300 ms test pulses to various potentials (from −100 to +60 mV in 10 mV increments). Holding potential, 0 mV (N = 5). ( B ) Mean current-voltage relationships for TRPV5-WT and TRPV5-682P channels (N = 5). These current-voltage relationships are similar to those reported for TRPV5 channels [62] . ( C ) Mean whole-cell tail currents measured in TRPV5-WT and TRPV5-682P injected Xenopus oocytes during test potentials applied in 10 mV increments from −70 to +40 mV after a pre-pulse to −100 mV in TRPV5-WT and TRPV5-682P channels (N = 5). ( D ) Time-dependent inhibition of TRPV-WT and TRPV5-682P whole-cell currents. Oocytes were stimulated every 1 s. The peak current amplitude was normalised to that recorded during the first pulse (N = 4). ( E ) Representative trace of Fura-2 ratio in HEK293 cells transiently transfected with an empty EGFP vector (mock), or EGFP-tagged TRPV5-WT or TRPV5-S682P. Cells expressing EGFP were selected and monitored for changes in intracellular Ca 2+ levels when extracellular Ca 2+ concentrations were varied from 1.4 mM Ca 2+ to 0 mM Ca 2+ (2 mM EDTA) and 1.4 mM Ca 2+ which was facilitated by superfusion. ( F ) Fura-2 levels under resting conditions (t0), minimal Fura-2 ratio after EDTA treatment (tmin) and peak level (tmax) upon administration of 1.4 mM Ca 2+ after EDTA treatment. Average data of cells transfected with the empty vector (n = 7), TRPV5-wt (n = 24) and TRPV5-S682P (N = 24) from at least three independent experiments. * p

    Article Snippet: For TRPV5 immuno-detection, 8- µm kidney cryosections were processed for immunofluorescence labelling as previously described ._ENREF_41 Kidney cryosections were co-stained with rabbit anti-TRPV5 (ACC-035, Alomone Labs, Jerusalem, Israel) and goat anti-AQP2 (sc-9882, Santa Cruz, Insight Biotechnology, Wembley, UK) polyclonal antibodies, or with goat anti-TRPV5 (sc-23379, Santa Cruz) and rabbit anti-NCC polyclonal antibodies, followed by the appropriate Alexa Fluor 488- or 594-conjugated secondary antibodies (Molecular Probes).

    Techniques: Mutagenesis, Injection, Mass Spectrometry, Inhibition, Transfection, Plasmid Preparation, Expressing

    Assessment of Renal Expression of Calcium Regulatory Genes and Proteins. Renal expression of ( A ) Trpv5 , ( B ) Trpv6 and ( C ) Cyp24a1 in wild-type (wt), Trpv5 682P/+ (het) and Trpv5 682P/682P (hom) mice (n = 6/group) were assessed by quantitative real-time PCR. All data were normalised to levels of the housekeeping gene Gapdh and wild-type values are expressed as 1. Histogram data are presented as mean ± SEM. # p

    Journal: PLoS ONE

    Article Title: Autosomal Dominant Hypercalciuria in a Mouse Model Due to a Mutation of the Epithelial Calcium Channel, TRPV5

    doi: 10.1371/journal.pone.0055412

    Figure Lengend Snippet: Assessment of Renal Expression of Calcium Regulatory Genes and Proteins. Renal expression of ( A ) Trpv5 , ( B ) Trpv6 and ( C ) Cyp24a1 in wild-type (wt), Trpv5 682P/+ (het) and Trpv5 682P/682P (hom) mice (n = 6/group) were assessed by quantitative real-time PCR. All data were normalised to levels of the housekeeping gene Gapdh and wild-type values are expressed as 1. Histogram data are presented as mean ± SEM. # p

    Article Snippet: For TRPV5 immuno-detection, 8- µm kidney cryosections were processed for immunofluorescence labelling as previously described ._ENREF_41 Kidney cryosections were co-stained with rabbit anti-TRPV5 (ACC-035, Alomone Labs, Jerusalem, Israel) and goat anti-AQP2 (sc-9882, Santa Cruz, Insight Biotechnology, Wembley, UK) polyclonal antibodies, or with goat anti-TRPV5 (sc-23379, Santa Cruz) and rabbit anti-NCC polyclonal antibodies, followed by the appropriate Alexa Fluor 488- or 594-conjugated secondary antibodies (Molecular Probes).

    Techniques: Expressing, Mouse Assay, Real-time Polymerase Chain Reaction

    Histological and immunohistochemical assessment of kidneys from HCALC1 mice. Representative images in Trpv5 +/+ (wt), Trpv5 682P/+ (het) and Trpv5 682P/682P (hom) mice of: ( A ) Masson's trichrome staining of renal cortex showing areas of interstitial fibrosis in Trpv5 682P/+ and Trpv5 682P/682P mice (light blue), ( B ) anti-CD3-labelling (green) showing a large number of T-lymphocytes present in the interstitial regions of the Trpv5 682P/+ and Trpv5 682P/682P mouse kidneys, ( C ) TUNEL-labelling (green) of the renal cortex showing the presence of tubular cell apoptosis in the Trpv5 682P/+ and Trpv5 682P/682P mouse kidneys. Scale bar = 50 µm. ( D ) Immunohistochemical images of kidney sections from wild-type (wt), Trpv5 682P/+ (het) and Trpv5 682P/682P (hom) mice, co-stained for TRPV5 (green) and NCC (red). * denotes co-localisation. Scale bar = 50 µm. ( E ) Kidney sections from wild-type (wt), Trpv5 682P/+ (het) and Trpv5 682P/682P (hom) mice, co-stained for TRPV5 (green) and AQP2 (red). * denotes co-localisation. Scale bar = 50 µm.

    Journal: PLoS ONE

    Article Title: Autosomal Dominant Hypercalciuria in a Mouse Model Due to a Mutation of the Epithelial Calcium Channel, TRPV5

    doi: 10.1371/journal.pone.0055412

    Figure Lengend Snippet: Histological and immunohistochemical assessment of kidneys from HCALC1 mice. Representative images in Trpv5 +/+ (wt), Trpv5 682P/+ (het) and Trpv5 682P/682P (hom) mice of: ( A ) Masson's trichrome staining of renal cortex showing areas of interstitial fibrosis in Trpv5 682P/+ and Trpv5 682P/682P mice (light blue), ( B ) anti-CD3-labelling (green) showing a large number of T-lymphocytes present in the interstitial regions of the Trpv5 682P/+ and Trpv5 682P/682P mouse kidneys, ( C ) TUNEL-labelling (green) of the renal cortex showing the presence of tubular cell apoptosis in the Trpv5 682P/+ and Trpv5 682P/682P mouse kidneys. Scale bar = 50 µm. ( D ) Immunohistochemical images of kidney sections from wild-type (wt), Trpv5 682P/+ (het) and Trpv5 682P/682P (hom) mice, co-stained for TRPV5 (green) and NCC (red). * denotes co-localisation. Scale bar = 50 µm. ( E ) Kidney sections from wild-type (wt), Trpv5 682P/+ (het) and Trpv5 682P/682P (hom) mice, co-stained for TRPV5 (green) and AQP2 (red). * denotes co-localisation. Scale bar = 50 µm.

    Article Snippet: For TRPV5 immuno-detection, 8- µm kidney cryosections were processed for immunofluorescence labelling as previously described ._ENREF_41 Kidney cryosections were co-stained with rabbit anti-TRPV5 (ACC-035, Alomone Labs, Jerusalem, Israel) and goat anti-AQP2 (sc-9882, Santa Cruz, Insight Biotechnology, Wembley, UK) polyclonal antibodies, or with goat anti-TRPV5 (sc-23379, Santa Cruz) and rabbit anti-NCC polyclonal antibodies, followed by the appropriate Alexa Fluor 488- or 594-conjugated secondary antibodies (Molecular Probes).

    Techniques: Immunohistochemistry, Mouse Assay, Staining, TUNEL Assay

    Hypercalciuria in HCALC1 ENU mutant mice and identification of a Trpv5 mutation. ( A ) Urine calcium/creatinine ratios in 23 G2 offspring of the HCALC1 founder male revealed that 10 of the 23 mice were hypercalciuric, consistent with an autosomal dominant inheritance. Bar, mean calcium/creatinine values. ( B ) Haplotype analysis of 89 G2 mice (39 hypercalciuric and 50 normocalciuric) was initially undertaken separately in the hypercalciuric and normocalciuric mice, as the penetrance of HCALC1 was unknown. Haplotype analysis of the hypercalciuric mice localised Hcalc1 to a 17.38 Mb interval on chromosome 6, flanked by rs13478688 and rs30110406 (broken double-headed arrow). Haplotype analysis using combined data for the hypercalciuric and normocalciuric mice identified the smaller interval, 11.94-Mb, flanked by rs13478709 and rs30110406 (solid double-headed arrow). The Hcalc1 locus is inherited with the C57BL/6J haplotype from the F1 founder male. Filled box, C57BL/6J allele; and open box, C3H/HeH allele. Number of mice observed for each haplotype is shown beneath each column. ( C ) DNA sequence analysis of Trpv5 identified a heterozygous T to C transition in codon 682 in hypercalciuric mice predicted to alter a wild-type serine (Ser) to a mutant proline (Pro). This mutation resulted in gain of a Bsa JI restriction enzyme site that was used to confirm the presence of the mutation in the 39 hypercalciuric mice (n = 3 shown) and its absence in the 50 normocalciuric mice (n = 3 shown). wt, wild-type; m, mutant. ( D ) Amino acid sequence alignment revealed evolutionary conservation of the wild-type mouse TRPV5 serine (S) residue at codon 682 (arrowed) in 5 species, as well as in mouse TRPV6 (mTrpv6). Identical residues are shaded black and conservative changes are shaded grey.

    Journal: PLoS ONE

    Article Title: Autosomal Dominant Hypercalciuria in a Mouse Model Due to a Mutation of the Epithelial Calcium Channel, TRPV5

    doi: 10.1371/journal.pone.0055412

    Figure Lengend Snippet: Hypercalciuria in HCALC1 ENU mutant mice and identification of a Trpv5 mutation. ( A ) Urine calcium/creatinine ratios in 23 G2 offspring of the HCALC1 founder male revealed that 10 of the 23 mice were hypercalciuric, consistent with an autosomal dominant inheritance. Bar, mean calcium/creatinine values. ( B ) Haplotype analysis of 89 G2 mice (39 hypercalciuric and 50 normocalciuric) was initially undertaken separately in the hypercalciuric and normocalciuric mice, as the penetrance of HCALC1 was unknown. Haplotype analysis of the hypercalciuric mice localised Hcalc1 to a 17.38 Mb interval on chromosome 6, flanked by rs13478688 and rs30110406 (broken double-headed arrow). Haplotype analysis using combined data for the hypercalciuric and normocalciuric mice identified the smaller interval, 11.94-Mb, flanked by rs13478709 and rs30110406 (solid double-headed arrow). The Hcalc1 locus is inherited with the C57BL/6J haplotype from the F1 founder male. Filled box, C57BL/6J allele; and open box, C3H/HeH allele. Number of mice observed for each haplotype is shown beneath each column. ( C ) DNA sequence analysis of Trpv5 identified a heterozygous T to C transition in codon 682 in hypercalciuric mice predicted to alter a wild-type serine (Ser) to a mutant proline (Pro). This mutation resulted in gain of a Bsa JI restriction enzyme site that was used to confirm the presence of the mutation in the 39 hypercalciuric mice (n = 3 shown) and its absence in the 50 normocalciuric mice (n = 3 shown). wt, wild-type; m, mutant. ( D ) Amino acid sequence alignment revealed evolutionary conservation of the wild-type mouse TRPV5 serine (S) residue at codon 682 (arrowed) in 5 species, as well as in mouse TRPV6 (mTrpv6). Identical residues are shaded black and conservative changes are shaded grey.

    Article Snippet: For TRPV5 immuno-detection, 8- µm kidney cryosections were processed for immunofluorescence labelling as previously described ._ENREF_41 Kidney cryosections were co-stained with rabbit anti-TRPV5 (ACC-035, Alomone Labs, Jerusalem, Israel) and goat anti-AQP2 (sc-9882, Santa Cruz, Insight Biotechnology, Wembley, UK) polyclonal antibodies, or with goat anti-TRPV5 (sc-23379, Santa Cruz) and rabbit anti-NCC polyclonal antibodies, followed by the appropriate Alexa Fluor 488- or 594-conjugated secondary antibodies (Molecular Probes).

    Techniques: Mutagenesis, Mouse Assay, Sequencing

    Microscopic images showing localization of TRPV5 in duck sperm. (A-B) Confocal microscopic images depicting the localization of TRPV5 (green) as detected by two different antibodies and Nucleus (blue) by DAPI. Mitochondria (red) is labelled by Mitotracker Red dye in A and C to highlight the channel expression in the mitochondrial region. ( C ) SR-SIM images of TRPV5 localization (using Ab1 antibody) at the head (left) and tail (right) of duck sperm is shown. ( D ) Zoomed up image of neck region of sperm depicting the absence of TRPV5 (green) in the neck region. The head (blue) and arrows mark the start and end point of mitochondrial region.

    Journal: bioRxiv

    Article Title: Differential expression and localization of thermosensitive Transient Receptor Potential Vanilloid (TRPV) channels in the mature sperm of white pekin duck (Anas platyrhynchos)

    doi: 10.1101/2020.02.10.941732

    Figure Lengend Snippet: Microscopic images showing localization of TRPV5 in duck sperm. (A-B) Confocal microscopic images depicting the localization of TRPV5 (green) as detected by two different antibodies and Nucleus (blue) by DAPI. Mitochondria (red) is labelled by Mitotracker Red dye in A and C to highlight the channel expression in the mitochondrial region. ( C ) SR-SIM images of TRPV5 localization (using Ab1 antibody) at the head (left) and tail (right) of duck sperm is shown. ( D ) Zoomed up image of neck region of sperm depicting the absence of TRPV5 (green) in the neck region. The head (blue) and arrows mark the start and end point of mitochondrial region.

    Article Snippet: Confocal microscopy using two different antibodies against the C-terminus of TRPV5 we found that TRPV5 is exclusively present at the tail (indicated by white arrows), with highest density immediately after the mitochondrial region and intensity gradually decreasing towards the tapering end of the tail ( ).

    Techniques: Expressing

    Microscopic images showing localization of TRPV6 in duck sperm. (A-B) Confocal microscopic images depicting the localization of TRPV6 (green) as detected by two different antibodies and Nucleus (blue) by DAPI. Mitochondria (red) is labelled by Mitotracker Red dye in A and C to highlight the channel expression in the mitochondrial region. ( C ) SR-SIM images of TRPV6 localization (using Ab1 antibody) at the head (left) and tail (right) of duck sperm is shown. ( D ) Zoomed up image of neck region of sperm depicting the absence of TRPV6 (green) in the neck region. The head (blue) and arrows mark the start and end point of mitochondrial region.

    Journal: bioRxiv

    Article Title: Differential expression and localization of thermosensitive Transient Receptor Potential Vanilloid (TRPV) channels in the mature sperm of white pekin duck (Anas platyrhynchos)

    doi: 10.1101/2020.02.10.941732

    Figure Lengend Snippet: Microscopic images showing localization of TRPV6 in duck sperm. (A-B) Confocal microscopic images depicting the localization of TRPV6 (green) as detected by two different antibodies and Nucleus (blue) by DAPI. Mitochondria (red) is labelled by Mitotracker Red dye in A and C to highlight the channel expression in the mitochondrial region. ( C ) SR-SIM images of TRPV6 localization (using Ab1 antibody) at the head (left) and tail (right) of duck sperm is shown. ( D ) Zoomed up image of neck region of sperm depicting the absence of TRPV6 (green) in the neck region. The head (blue) and arrows mark the start and end point of mitochondrial region.

    Article Snippet: Confocal microscopy using two different antibodies against the C-terminus of TRPV5 we found that TRPV5 is exclusively present at the tail (indicated by white arrows), with highest density immediately after the mitochondrial region and intensity gradually decreasing towards the tapering end of the tail ( ).

    Techniques: Expressing

    Endogenous expression of TRPV channels in duck sperm. Western blot analysis of duck sperm extracts probed with different TRPV-specific antibodies are shown. A. TRPV1 specific band is detected by a specific antibody (directed against the C-terminus of TRPV1, Alomone Labs) in absence but not in presence of its blocking peptide; B. Western blot analysis with antibody that detects TRPV2 (raised against the C-terminus, Alomone Labs). C. Two different antibodies detecting TRPV3 [raised against the C-terminus, (Ab1: Alomone Labs) and N-terminus (Ab3: Sigma Aldrich)] detect similar expression pattern of TRPV3. D. Two different antibodies raised against the TRPV4 [raised against C-terminus, Ab1: Alomone Labs) and N-terminus (Ab3: Sigma Aldrich)] detect TRPV4 at the expected size. E. Two different antibodies raised against the C-terminus of TRPV5 (Ab1: Alomone Labs and Ab2: Sigma-Aldrich) detects TRPV5 at expected size. F. A specific antibody raised against the C-terminus of TRPV6 (Ab-1: Alomone Labs) detects TRPV6 in absence but not in presence of its blocking peptide.

    Journal: bioRxiv

    Article Title: Differential expression and localization of thermosensitive Transient Receptor Potential Vanilloid (TRPV) channels in the mature sperm of white pekin duck (Anas platyrhynchos)

    doi: 10.1101/2020.02.10.941732

    Figure Lengend Snippet: Endogenous expression of TRPV channels in duck sperm. Western blot analysis of duck sperm extracts probed with different TRPV-specific antibodies are shown. A. TRPV1 specific band is detected by a specific antibody (directed against the C-terminus of TRPV1, Alomone Labs) in absence but not in presence of its blocking peptide; B. Western blot analysis with antibody that detects TRPV2 (raised against the C-terminus, Alomone Labs). C. Two different antibodies detecting TRPV3 [raised against the C-terminus, (Ab1: Alomone Labs) and N-terminus (Ab3: Sigma Aldrich)] detect similar expression pattern of TRPV3. D. Two different antibodies raised against the TRPV4 [raised against C-terminus, Ab1: Alomone Labs) and N-terminus (Ab3: Sigma Aldrich)] detect TRPV4 at the expected size. E. Two different antibodies raised against the C-terminus of TRPV5 (Ab1: Alomone Labs and Ab2: Sigma-Aldrich) detects TRPV5 at expected size. F. A specific antibody raised against the C-terminus of TRPV6 (Ab-1: Alomone Labs) detects TRPV6 in absence but not in presence of its blocking peptide.

    Article Snippet: Confocal microscopy using two different antibodies against the C-terminus of TRPV5 we found that TRPV5 is exclusively present at the tail (indicated by white arrows), with highest density immediately after the mitochondrial region and intensity gradually decreasing towards the tapering end of the tail ( ).

    Techniques: Expressing, Western Blot, Blocking Assay