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Addgene inc pme sema7a
Pme Sema7a, supplied by Addgene inc, 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/product/pme+sema7a/pmc11318972-287-17-22?v=Addgene+inc
Average 93 stars, based on 1 article reviews

pme sema7a - by Bioz Stars, 2026-07

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(A) A volumetric rendering of a PLL neuromast depicts the sensory axons (magenta) that branch from the lateral line nerve to arborize around the basolateral surface of the hair cell cluster (green). Additional cell types in the neuromast are not labeled. Larval age, 3 dpf. (B) A schematic drawing of the two variants of the Sema7A protein molecule depicts the full-length GPI-anchored form and the smaller, potentially secreted form. Both the molecules include a signal sequence (green) and a conserved sema domain (orange). Gel-based RT-PCR analysis indicates the presence of both the variants in developing larva. (C) A surface micrograph of a neuromast at 3 dpf depicts two pairs of mature hair cells (green dashed lines) and a pair of immature hair cells (grey dashed lines). Inset: among the three pairs of hair cell apices, the immature pair is indicated by arrowheads. Immunolabeling reveals that the Sema7A protein (orange) occurs consistently at the subapical region (arrowheads) and at the basolateral surface (arrows) of a hair cell. In this and in each of the subsequent neuromast images, anterior is to the left and ventral to the top. (D) A plot quantifies developmental changes in the average Sema7A intensity in both rostrally and caudally polarized hair cells of neuromasts from 1.5 dpf to 4 dpf. The data stem from 18, 36, 39, and 40 hair cells in neuromasts of respectively 1.5 dpf, 2 dpf, 3 dpf, and 4 dpf larvae. (E) A plot quantifies the distribution of average Sema7A intensity along the hair cell’s apicobasal axis. The results stem from 52, 57, and 54 hair cells of neuromasts from 2 dpf, 3 dpf, and 4 dpf larvae. (F) An immunofluorescence image at the nuclear level of a 4 dpf neuromast shows the contact of the sensory arbors (magenta) with the basolateral surface of the hair cells (green). Immunolabeling for Sema7A (orange) reveals enrichment of the protein at the hair cell bases and sensory-axon interfaces (arrowheads). HC, hair cell; Scale bars, 5 μ m; au, arbitrary unit; means ± SEMs; *** implies p < 0.001.

Journal: bioRxiv

Article Title: Semaphorin7A patterns neural circuitry in the lateral line of the zebrafish

doi: 10.1101/2022.11.17.516962

Figure Lengend Snippet: (A) A volumetric rendering of a PLL neuromast depicts the sensory axons (magenta) that branch from the lateral line nerve to arborize around the basolateral surface of the hair cell cluster (green). Additional cell types in the neuromast are not labeled. Larval age, 3 dpf. (B) A schematic drawing of the two variants of the Sema7A protein molecule depicts the full-length GPI-anchored form and the smaller, potentially secreted form. Both the molecules include a signal sequence (green) and a conserved sema domain (orange). Gel-based RT-PCR analysis indicates the presence of both the variants in developing larva. (C) A surface micrograph of a neuromast at 3 dpf depicts two pairs of mature hair cells (green dashed lines) and a pair of immature hair cells (grey dashed lines). Inset: among the three pairs of hair cell apices, the immature pair is indicated by arrowheads. Immunolabeling reveals that the Sema7A protein (orange) occurs consistently at the subapical region (arrowheads) and at the basolateral surface (arrows) of a hair cell. In this and in each of the subsequent neuromast images, anterior is to the left and ventral to the top. (D) A plot quantifies developmental changes in the average Sema7A intensity in both rostrally and caudally polarized hair cells of neuromasts from 1.5 dpf to 4 dpf. The data stem from 18, 36, 39, and 40 hair cells in neuromasts of respectively 1.5 dpf, 2 dpf, 3 dpf, and 4 dpf larvae. (E) A plot quantifies the distribution of average Sema7A intensity along the hair cell’s apicobasal axis. The results stem from 52, 57, and 54 hair cells of neuromasts from 2 dpf, 3 dpf, and 4 dpf larvae. (F) An immunofluorescence image at the nuclear level of a 4 dpf neuromast shows the contact of the sensory arbors (magenta) with the basolateral surface of the hair cells (green). Immunolabeling for Sema7A (orange) reveals enrichment of the protein at the hair cell bases and sensory-axon interfaces (arrowheads). HC, hair cell; Scale bars, 5 μ m; au, arbitrary unit; means ± SEMs; *** implies p < 0.001.

Article Snippet: To generate the hsp70:sema7a sec -mKate2 construct, gateway cloning was performed by combining the plasmids p5E-hsp70 , pME-sema7a sec , p3E-mKate2-myc no-pA (Addgene, 80812), and pDESTtol2pACrymCherry (Addgene, 64023) with LR Clonase II Plus (Invitrogen).

Techniques: Labeling, Sequencing, Reverse Transcription Polymerase Chain Reaction, Immunolabeling, Immunofluorescence

(A) Distinct sets of primers (arrows) identify the two variants of the Sema7A protein molecule. (B-D) Surface micrographs show hair cells marked with actin-GFP (green dashed lines) and the localization of Sema7A protein (orange) in the subapical (arrowheads) and basal regions (arrows) of the corresponding hair cells from neuromasts of developing control larvae. The inset in panel B depicts the apices of the immature hair cell pair. (E) Immunolabeling with only the secondary antibody fails to detect any Sema7A signal. (F) A schematic diagram of a single hair cell depicts the three distinct regions along the apicobasal axis of the cell. Sema7A intensity was measured along this apicobasal axis (red dashed line). (G). A surface micrograph depicts hair cells (green, left) whose subapical region (red dashed line) harbors the Golgi network (cyan, middle) where the Sema7A protein is enriched (orange, right). Scale bars, 5 μ m.

Journal: bioRxiv

Article Title: Semaphorin7A patterns neural circuitry in the lateral line of the zebrafish

doi: 10.1101/2022.11.17.516962

Figure Lengend Snippet: (A) Distinct sets of primers (arrows) identify the two variants of the Sema7A protein molecule. (B-D) Surface micrographs show hair cells marked with actin-GFP (green dashed lines) and the localization of Sema7A protein (orange) in the subapical (arrowheads) and basal regions (arrows) of the corresponding hair cells from neuromasts of developing control larvae. The inset in panel B depicts the apices of the immature hair cell pair. (E) Immunolabeling with only the secondary antibody fails to detect any Sema7A signal. (F) A schematic diagram of a single hair cell depicts the three distinct regions along the apicobasal axis of the cell. Sema7A intensity was measured along this apicobasal axis (red dashed line). (G). A surface micrograph depicts hair cells (green, left) whose subapical region (red dashed line) harbors the Golgi network (cyan, middle) where the Sema7A protein is enriched (orange, right). Scale bars, 5 μ m.

Techniques: Immunolabeling

(A) Micrographs depict the overall morphologies and sema7a genomic profiles of 2 dpf control (top) and sema7a -/- larvae (bottom). (B) In a surface micrograph of a 3 dpf control neuromast, myosin VI (green) marks hair cells (green dashed lines). Sema7A (orange) protein is highly enriched in the subapical region (arrowheads) and basolateral (arrow) region of the hair cells. (C,D) Micrographs depict combined 4 dpf hair cell clusters, each of whose centers arise located at (0,0). The X- and Y-coordinates represent the anteroposterior (AP) and dorsoventral (DV) axes of the larva, respectively. Positive values of the X- and Y-coordinates represent posterior and ventral directions, respectively. Combined hair cell clusters from 27 control and 27 sema7a -/- mutant neuromasts are represented. (E,F) Micrographs depict the skeletonized networks of the combined 2 dpf hair cell clusters from 17 control and 17 sema7a -/- mutant neuromasts. (G,H) Micrographs depict the skeletonized networks of the combined 3 dpf hair cell clusters from 29 control and 29 sema7a -/- mutant neuromasts. (I) A plot quantitates the densities of the sensory arbors around the center of the combined hair cell clusters for 33 control (magenta) and 17 sema7a -/- (red) neuromasts at 2 dpf. The shaded area marks the region proximal to the boundary of the combined hair cell cluster. (J) A plot quantitates the densities of the sensory arbors around the center of the combined hair cell clusters for 29 control (magenta) and 53 sema7a -/- (red) neuromasts at 3 dpf. The shaded area marks the region proximal to the boundary of the combined hair cell cluster. Scale bar, 5 μ m; au, arbitrary units.

Journal: bioRxiv

Article Title: Semaphorin7A patterns neural circuitry in the lateral line of the zebrafish

doi: 10.1101/2022.11.17.516962

Figure Lengend Snippet: (A) Micrographs depict the overall morphologies and sema7a genomic profiles of 2 dpf control (top) and sema7a -/- larvae (bottom). (B) In a surface micrograph of a 3 dpf control neuromast, myosin VI (green) marks hair cells (green dashed lines). Sema7A (orange) protein is highly enriched in the subapical region (arrowheads) and basolateral (arrow) region of the hair cells. (C,D) Micrographs depict combined 4 dpf hair cell clusters, each of whose centers arise located at (0,0). The X- and Y-coordinates represent the anteroposterior (AP) and dorsoventral (DV) axes of the larva, respectively. Positive values of the X- and Y-coordinates represent posterior and ventral directions, respectively. Combined hair cell clusters from 27 control and 27 sema7a -/- mutant neuromasts are represented. (E,F) Micrographs depict the skeletonized networks of the combined 2 dpf hair cell clusters from 17 control and 17 sema7a -/- mutant neuromasts. (G,H) Micrographs depict the skeletonized networks of the combined 3 dpf hair cell clusters from 29 control and 29 sema7a -/- mutant neuromasts. (I) A plot quantitates the densities of the sensory arbors around the center of the combined hair cell clusters for 33 control (magenta) and 17 sema7a -/- (red) neuromasts at 2 dpf. The shaded area marks the region proximal to the boundary of the combined hair cell cluster. (J) A plot quantitates the densities of the sensory arbors around the center of the combined hair cell clusters for 29 control (magenta) and 53 sema7a -/- (red) neuromasts at 3 dpf. The shaded area marks the region proximal to the boundary of the combined hair cell cluster. Scale bar, 5 μ m; au, arbitrary units.

Techniques: Mutagenesis

(A) In a micrograph of a 3 dpf sema7a -/- neuromast, Myosin VI (green) marks the hair cells (green dashed lines) in which the level of Sema7A (orange) is highly reduced, with sporadic localization in the subapical region (arrowheads) and none in the basolateral (arrow) region. A plot of normalized Sema7A intensity from 99 control and 100 sema7a -/- hair cells quantitates the effect. (B,C) Surface views of a control and a sema7a -/- neuromast at 4 dpf depict the interaction of the sensory arbors (magenta) with hair cell clusters (green). In the control, the arbors intimately contact the hair cells, with a few exceptions (arrowhead). In the sema7a -/- mutant, the arbors direct many aberrant projections (arrowheads) away from the hair cell cluster. The two immature hair cell pairs in the sema7a -/- neuromast are indicated by arrows. (D,E) Skeletonized networks portray the three-dimensional topology of the sensory arbors from the control and the sema7a -/- neuromasts depicted in panels B and C. The pseudocolored trajectories depict the increase in arbor contour length from each point of arborization, defined as the point at which the lateral line branch (magenta) contacts hair cell cluster. (F,G) Micrographs depict the skeletonized networks of the combined 4 dpf hair cell clusters, whose centers are located at (0,0). The X- and Y-coordinates represent the anteroposterior (AP) and the dorsoventral (DV) axes of the larva, respectively. Positive values of the X- and Y-coordinates represent the posterior and ventral directions, respectively. Combined skeletonized network traces from 27 control and 27 sema7a -/- mutant neuromasts are represented. H) The plot denotes the densities of the sensory arbors around the center of the combined hair cell clusters for 35 control (magenta) and 27 sema7a -/- (red) neuromasts at 4 dpf. The shaded area marks the region proximal to the boundary of the combined hair cell cluster. (I) The plot quantifies the degree of contact of the sensory arbors to their hair cell clusters in individual neuromasts from both control (magenta) and sema7a -/- mutants (red), each point represents a single neuromast. Thirty-three, 29, and 35 neuromasts were analyzed from 2 dpf, 3 dpf, and 4 dpf control larvae, respectively. Seventeen, 53, and 27 neuromasts were analyzed from 2 dpf, 3 dpf, and 4 dpf sema7a -/- mutant larvae, respectively. HC, hair cell; Scale bars, 5 μ m; au, arbitrary unit; means ± SEMs; *** implies p < 0.001; ** signifies p < 0.01.

Journal: bioRxiv

Article Title: Semaphorin7A patterns neural circuitry in the lateral line of the zebrafish

doi: 10.1101/2022.11.17.516962

Figure Lengend Snippet: (A) In a micrograph of a 3 dpf sema7a -/- neuromast, Myosin VI (green) marks the hair cells (green dashed lines) in which the level of Sema7A (orange) is highly reduced, with sporadic localization in the subapical region (arrowheads) and none in the basolateral (arrow) region. A plot of normalized Sema7A intensity from 99 control and 100 sema7a -/- hair cells quantitates the effect. (B,C) Surface views of a control and a sema7a -/- neuromast at 4 dpf depict the interaction of the sensory arbors (magenta) with hair cell clusters (green). In the control, the arbors intimately contact the hair cells, with a few exceptions (arrowhead). In the sema7a -/- mutant, the arbors direct many aberrant projections (arrowheads) away from the hair cell cluster. The two immature hair cell pairs in the sema7a -/- neuromast are indicated by arrows. (D,E) Skeletonized networks portray the three-dimensional topology of the sensory arbors from the control and the sema7a -/- neuromasts depicted in panels B and C. The pseudocolored trajectories depict the increase in arbor contour length from each point of arborization, defined as the point at which the lateral line branch (magenta) contacts hair cell cluster. (F,G) Micrographs depict the skeletonized networks of the combined 4 dpf hair cell clusters, whose centers are located at (0,0). The X- and Y-coordinates represent the anteroposterior (AP) and the dorsoventral (DV) axes of the larva, respectively. Positive values of the X- and Y-coordinates represent the posterior and ventral directions, respectively. Combined skeletonized network traces from 27 control and 27 sema7a -/- mutant neuromasts are represented. H) The plot denotes the densities of the sensory arbors around the center of the combined hair cell clusters for 35 control (magenta) and 27 sema7a -/- (red) neuromasts at 4 dpf. The shaded area marks the region proximal to the boundary of the combined hair cell cluster. (I) The plot quantifies the degree of contact of the sensory arbors to their hair cell clusters in individual neuromasts from both control (magenta) and sema7a -/- mutants (red), each point represents a single neuromast. Thirty-three, 29, and 35 neuromasts were analyzed from 2 dpf, 3 dpf, and 4 dpf control larvae, respectively. Seventeen, 53, and 27 neuromasts were analyzed from 2 dpf, 3 dpf, and 4 dpf sema7a -/- mutant larvae, respectively. HC, hair cell; Scale bars, 5 μ m; au, arbitrary unit; means ± SEMs; *** implies p < 0.001; ** signifies p < 0.01.

Techniques: Mutagenesis

(A) A diagrammatic overview depicts the generation of a transgenic animal that expresses the Sema7A sec protein ectopically under the control of a thermally inducible promoter. Larvae with ectopic myotomal-integration near the network of sensory arbors (red line) were imaged to analyze arbor morphology. (B) A schematic drawing of a sensory arbor from a heat-shocked larva depicts an extended axonal projection (arrowhead) that reaches toward the myofibers expressing the ectopic Sema7A sec protein. Parameters that quantitate the accuracy of the extended axonal projections toward the ectopic Sema7A sec sources are denoted. (C) In a micrograph of an ectopically expressing Sema7A sec (orange) larva, the sensory arbor (magenta) extends two aberrant axonal projections. One elongates (cyan arrowhead) along the somite boundary to reach and contact an ectopically integrated muscle progenitor cell (white dashed line) and the other (red arrowhead) reenters the posterior lateral line nerve while following a second ectopic source. The through-focus scan (i–iv) from the epidermis to the dermomyotome reveals the intimate contact between the aberrant sensory arbor (arrowheads) and the muscle progenitor cell. MPC, muscle progenitor cell. (D) In a micrograph of an ectopically expressing Sema7A sec (orange) larva, the sensory arbor (magenta) extends a single aberrant axonal process (cyan arrowhead) to reach ectopically integrated myofibers (white dashed line). The through-focus scan (i–iv) from the epidermis to the deep myotome reveals the proximal association of the aberrant sensory arbor (arrowheads) to the myofibers. Melanocytes (yellow arrowhead) along the horizontal myoseptum intermittently block the visibility of the lateral line nerve. (E) An injected, but not heat-shocked, control larva does not express ectopic Sema7A sec and does not show aberrant projection from the sensory arbor. (F) A plot demonstrates the accuracy of 18 extended axonal arbors in finding ectopic Sema7A sec sources. Each circle represents a single ectopic integration event. The two pairs of numbers represent the minimal and maximal lengths of the projection path (black) and its corresponding source path (green). (G) A plot quantitates the distribution of projection-proximity length from 18 ectopic integration events. Scale bars, 20 μ m; means ± SEMs.

Journal: bioRxiv

Article Title: Semaphorin7A patterns neural circuitry in the lateral line of the zebrafish

doi: 10.1101/2022.11.17.516962

Figure Lengend Snippet: (A) A diagrammatic overview depicts the generation of a transgenic animal that expresses the Sema7A sec protein ectopically under the control of a thermally inducible promoter. Larvae with ectopic myotomal-integration near the network of sensory arbors (red line) were imaged to analyze arbor morphology. (B) A schematic drawing of a sensory arbor from a heat-shocked larva depicts an extended axonal projection (arrowhead) that reaches toward the myofibers expressing the ectopic Sema7A sec protein. Parameters that quantitate the accuracy of the extended axonal projections toward the ectopic Sema7A sec sources are denoted. (C) In a micrograph of an ectopically expressing Sema7A sec (orange) larva, the sensory arbor (magenta) extends two aberrant axonal projections. One elongates (cyan arrowhead) along the somite boundary to reach and contact an ectopically integrated muscle progenitor cell (white dashed line) and the other (red arrowhead) reenters the posterior lateral line nerve while following a second ectopic source. The through-focus scan (i–iv) from the epidermis to the dermomyotome reveals the intimate contact between the aberrant sensory arbor (arrowheads) and the muscle progenitor cell. MPC, muscle progenitor cell. (D) In a micrograph of an ectopically expressing Sema7A sec (orange) larva, the sensory arbor (magenta) extends a single aberrant axonal process (cyan arrowhead) to reach ectopically integrated myofibers (white dashed line). The through-focus scan (i–iv) from the epidermis to the deep myotome reveals the proximal association of the aberrant sensory arbor (arrowheads) to the myofibers. Melanocytes (yellow arrowhead) along the horizontal myoseptum intermittently block the visibility of the lateral line nerve. (E) An injected, but not heat-shocked, control larva does not express ectopic Sema7A sec and does not show aberrant projection from the sensory arbor. (F) A plot demonstrates the accuracy of 18 extended axonal arbors in finding ectopic Sema7A sec sources. Each circle represents a single ectopic integration event. The two pairs of numbers represent the minimal and maximal lengths of the projection path (black) and its corresponding source path (green). (G) A plot quantitates the distribution of projection-proximity length from 18 ectopic integration events. Scale bars, 20 μ m; means ± SEMs.

Techniques: Transgenic Assay, Expressing, Blocking Assay, Injection

(A,B) Maximal-intensity projections of micrographs from a control and a sema7a -/- neuromast depict the presynaptic aggregates marked by CTBP (cyan). The approximate hair cell basal region is outlined by dashed green circles. (C-F) Plots quantitate the numbers of presynaptic aggregates (C,D) and their areas (E,F), in each hair cell across developmental stages. One hundred and fifty-two, 317, and 325 hair cells were analyzed from 2 dpf, 3 dpf, and 4 dpf control larvae, respectively. One hundred and sixty, 216, and 177 hair cells were analyzed from 2 dpf, 3 dpf, and 4 dpf sema7a -/- mutant larvae, respectively. (I,J) Maximal-intensity projections from micrographs of a control and a sema7a -/- neuromast depict the postsynaptic aggregates marked by MAGUK (yellow). The approximate hair cell basal region is outlined by dashed green circles. (K,N) Plots quantitate the numbers of postsynaptic aggregate (K,L) and their areas (M,N), in each hair cell across developmental stages. One hundred and fifty, 218, and 167 hair cells were analyzed from 2 dpf, 3 dpf, and 4 dpf control larvae, respectively. Ninety-seven, 141, and 141 hair cells were analyzed from 2 dpf, 3 dpf, and 4 dpf sema7a -/- mutant larvae, respectively. Scale bars, 5 μ m.

Journal: bioRxiv

Article Title: Semaphorin7A patterns neural circuitry in the lateral line of the zebrafish

doi: 10.1101/2022.11.17.516962

Figure Lengend Snippet: (A,B) Maximal-intensity projections of micrographs from a control and a sema7a -/- neuromast depict the presynaptic aggregates marked by CTBP (cyan). The approximate hair cell basal region is outlined by dashed green circles. (C-F) Plots quantitate the numbers of presynaptic aggregates (C,D) and their areas (E,F), in each hair cell across developmental stages. One hundred and fifty-two, 317, and 325 hair cells were analyzed from 2 dpf, 3 dpf, and 4 dpf control larvae, respectively. One hundred and sixty, 216, and 177 hair cells were analyzed from 2 dpf, 3 dpf, and 4 dpf sema7a -/- mutant larvae, respectively. (I,J) Maximal-intensity projections from micrographs of a control and a sema7a -/- neuromast depict the postsynaptic aggregates marked by MAGUK (yellow). The approximate hair cell basal region is outlined by dashed green circles. (K,N) Plots quantitate the numbers of postsynaptic aggregate (K,L) and their areas (M,N), in each hair cell across developmental stages. One hundred and fifty, 218, and 167 hair cells were analyzed from 2 dpf, 3 dpf, and 4 dpf control larvae, respectively. Ninety-seven, 141, and 141 hair cells were analyzed from 2 dpf, 3 dpf, and 4 dpf sema7a -/- mutant larvae, respectively. Scale bars, 5 μ m.

Techniques: Mutagenesis

(A-F) The plots quantitate the average numbers and areas of presynaptic aggregates from control and sema7a -/- neuromasts. Significant decreases in the numbers and areas of presynaptic aggregates are observed across development. Three hundred and forty-one, 1229, and 1286 presynaptic aggregates were analyzed from 2 dpf, 3 dpf, and 4 dpf control larvae, respectively. Two hundred and thirty-five, 643, and 419 presynaptic aggregates were analyzed from 2 dpf, 3 dpf, and 4 dpf sema7a -/- mutant larvae, respectively. (G-L) The plots quantitate the average numbers and areas of postsynaptic aggregates from control and sema7a -/- neuromasts. Significant decreases in the numbers and areas of postsynaptic aggregates occurred during development. Three hundred and ninety-seven, 1243, and 1300 postsynaptic aggregates were analyzed from 2 dpf, 3 dpf, and 4 dpf control larvae, respectively. Three hundred and four, 651, and 451 postsynaptic aggregates were analyzed from 2 dpf, 3 dpf, and 4 dpf sema7a -/- mutant larvae, respectively. Means ± SEMs; *** implies p < 0.001.

Journal: bioRxiv

Article Title: Semaphorin7A patterns neural circuitry in the lateral line of the zebrafish

doi: 10.1101/2022.11.17.516962

Figure Lengend Snippet: (A-F) The plots quantitate the average numbers and areas of presynaptic aggregates from control and sema7a -/- neuromasts. Significant decreases in the numbers and areas of presynaptic aggregates are observed across development. Three hundred and forty-one, 1229, and 1286 presynaptic aggregates were analyzed from 2 dpf, 3 dpf, and 4 dpf control larvae, respectively. Two hundred and thirty-five, 643, and 419 presynaptic aggregates were analyzed from 2 dpf, 3 dpf, and 4 dpf sema7a -/- mutant larvae, respectively. (G-L) The plots quantitate the average numbers and areas of postsynaptic aggregates from control and sema7a -/- neuromasts. Significant decreases in the numbers and areas of postsynaptic aggregates occurred during development. Three hundred and ninety-seven, 1243, and 1300 postsynaptic aggregates were analyzed from 2 dpf, 3 dpf, and 4 dpf control larvae, respectively. Three hundred and four, 651, and 451 postsynaptic aggregates were analyzed from 2 dpf, 3 dpf, and 4 dpf sema7a -/- mutant larvae, respectively. Means ± SEMs; *** implies p < 0.001.

Techniques: Mutagenesis

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