Journal: Frontiers in Neuroanatomy

Article Title: Asymmetric Distributions of Achromatic Bipolar Cells in the Mouse Retina

doi: 10.3389/fnana.2021.786142

Figure Lengend Snippet: Primary antibodies.

Article Snippet: HCN4 , GST fusion protein with amino acids 119-155 of human HCN4 , Alomone Labs, APC-052, Rabbit polyclonal , AB_2039906 , 1:500.

Techniques: Recombinant

Journal: Frontiers in Neuroanatomy

Article Title: Asymmetric Distributions of Achromatic Bipolar Cells in the Mouse Retina

doi: 10.3389/fnana.2021.786142

Figure Lengend Snippet: The Rik mouse GFP-expressing cells were type 2 and 3a bipolar cells. Images were taken from a single z section. (A) A type 2 bipolar cell marker, Syt2, colocalized with GFP-expressing cells. Asterisks indicate somas labeled both with Syt2 and GFP. (B) A type 3a marker, HCN4, colocalized with GFP cells. Asterisks indicate colocalized cells. (C) A type 3b marker, PKARIIβ, did not colocalize with GFP cells. (D) A type 4 marker, Csen, did not colocalize with GFP cells. (E) A rod bipolar cell marker, PKCα, did not colocalized with GFP cells.

Article Snippet: HCN4 , GST fusion protein with amino acids 119-155 of human HCN4 , Alomone Labs, APC-052, Rabbit polyclonal , AB_2039906 , 1:500.

Techniques: Expressing, Marker, Labeling

Journal: Frontiers in Neuroanatomy

Article Title: Asymmetric Distributions of Achromatic Bipolar Cells in the Mouse Retina

doi: 10.3389/fnana.2021.786142

Figure Lengend Snippet: The Lhx4 mouse GFP-expressing cells were type 3b and 4 bipolar cells. Images were taken from a single z section. (A) A type 2 bipolar cell marker, Syt2, did not colocalize with GFP-expressing cells. (B) A type 3a marker, HCN4, did not colocalize with GFP cells. (C) A type 3b marker, PKARIIβ, colocalized with GFP cells, indicated by asterisks. (D) A type 4 marker, Csen, colocalized with GFP cells, indicated by asterisks. (E) A rod bipolar cell marker, PKCα, did not colocalize with GFP cells.

Article Snippet: HCN4 , GST fusion protein with amino acids 119-155 of human HCN4 , Alomone Labs, APC-052, Rabbit polyclonal , AB_2039906 , 1:500.

Techniques: Expressing, Marker

Journal: Frontiers in Neuroanatomy

Article Title: Asymmetric Distributions of Achromatic Bipolar Cells in the Mouse Retina

doi: 10.3389/fnana.2021.786142

Figure Lengend Snippet: Bipolar cell type-dependent asymmetricity in the Rik mouse. (A) Subsets of GFP cells were colocalized with Syt2 and HCN4 antibodies, which revealed three different types of bipolar cells in the wholemount retinal tissues. (B) The density of Syt2-positive GFP cells did not show asymmetric distributions along the nasal-temporal axis (linear regression analysis). Each color on the graphs in B-D represent a different mouse. (C) The density of HCN4-positive, type 3a cells was higher along with the nasal-temporal axis (linear regression analysis). (D) The density of GFP+/Syt2-/ HCN4-, type 6 bipolar cells, was found to show a gradient of decreasing density moving from the nasal region to the temporal region (linear regression analysis).

Article Snippet: HCN4 , GST fusion protein with amino acids 119-155 of human HCN4 , Alomone Labs, APC-052, Rabbit polyclonal , AB_2039906 , 1:500.

Techniques:

Journal: The Journal of comparative neurology

Article Title: DSCAM Localization and Function at the Mouse Cone Synapse

doi: 10.1002/cne.23552

Figure Lengend Snippet: Primary Antibodies

Article Snippet: 1:2,000 HCN4 Amino acids 119–155 of human HCN4 (HGHLHDSAEERRLIAEGDASPGEDRTPPGLAAEPERP) Alomone, Jerusalem, Israel.

Techniques: Recombinant, Purification, Transduction

Journal: The Journal of comparative neurology

Article Title: DSCAM Localization and Function at the Mouse Cone Synapse

doi: 10.1002/cne.23552

Figure Lengend Snippet: A–D, Sections of adult (>P42) wild type and Dscam2J/2J retina were stained with antibodies to NK3R to label type 1 and type 2 cone bipolar cells, and PNA (N=4 retinas). Differences when comparing wild type and Dscam loss of function retina were not detected. E–F, Sections of adult (>P42) wild type and Dscam2J/2J retina were stained with antibodies to recoverin to label type 2 cone bipolar cells and rods, and PNA (N=4 retinas). Differences when comparing wild type and Dscam loss of function retina were not detected. I–L, Sections of adult (>P42) wild type and Dscam2J/2J retina were stained with HCN4 to label type 3a cone bipolar cells and PNA (N=4 retinas). Lamination of HCN4 positive neurites in the ON half of the inner plexiform layer are longer in the Dscam2J/2J retina compared to wild type controls (I versus J; arrow). No differences were detected in the outer plexiform layer. M–T, Sections of adult (>P42) wild type and Dscam2J/2J retina were stained with antibodies to PKARIIβ to label type 3b cone bipolar cells and PNA (N>4 retinas). PKARIIβ immunopositive bipolar cells have clumped dendrite arbors proximal to cone pedicles, compared to wild type (P, S and T; mutant vs. O, Q and R; wild type). The scale bar in (A) is equivalent to 100 μm in A, B, E, F, I, J, M and N. The scale bar in (C) is equivalent to 20 μm in C, D, G, H, K, L, O and P. The scale bar in (Q) is equivalent to 10 μm in Q–T.

Article Snippet: 1:2,000 HCN4 Amino acids 119–155 of human HCN4 (HGHLHDSAEERRLIAEGDASPGEDRTPPGLAAEPERP) Alomone, Jerusalem, Israel.

Techniques: Staining, Mutagenesis

Journal: The Journal of comparative neurology

Article Title: DSCAM Localization and Function at the Mouse Cone Synapse

doi: 10.1002/cne.23552

Figure Lengend Snippet: A, Section of DscamFD/FD retina stained with antibodies to DSCAM and Brn3A (N=2). DSCAM immunoreactivity was observed in the soma of Brn3A positive cells in the retinal ganglion cell layer. B, Section of DscamFD/FD retina stained with antibodies to DSCAM and disabled (Dab) to stain AII amacrine cells (N=2). AII amacrine cells did not contain DSCAM in their soma. C, Section of DscamFD/FD retina stained with antibodies to DSCAM and ChAT, to label cholinergic amacrine cells (N=2). Cholinergic amacrine cell soma in the inner nuclear layer (top) or retinal ganglion layer (bottom) did not contain DSCAM protein. D, Section of DscamFD/FD retina stained with antibodies to DSCAM and bNOS (N=2). bNOS positive amacrine cells in both the inner nuclear layer (top) and retinal ganglion cell layer (bottom) contained DSCAM protein in their soma. E, Section of DscamFD/FD retina stained with antibodies to DSCAM and Chx10, to label bipolar cells (N=4). DSCAM immunoreactivity was observed in the soma surrounding some Chx10 positive cells. F and G, Section of DscamFD/FD retina stained with antibodies to HCN4 to label a subset of amacrine cells (G) and type 3a cone bipolar cells (F) (N=2 retinas). F, HCN4-positive bipolar cells, distinguished from amacrine cells by projection of dendrites to the outer plexiform layer and location, contained DSCAM protein in their soma. G, HCN4-positive amacrine cells, identified by their location proximal to the inner plexiform layer, did not contain DSCAM protein. H, Sections of DscamFD/FD retina were stained with antibodies to DSCAM and PKARIIβ to label type 3b bipolar cells. DSCAM immunoreactivity was observed in PKARIIβ positive cells. I, Sections of DscamFD/FD retina were stained with antibodies to DSCAM and Syt2, to label type 2 and type 6 bipolar cells (N=2 retinas). Some Syt2 immunopositive cells contained DSCAM immunoreactivity while others did not. J, Sections of DscamFD/FD retina were stained with antibodies to DSCAM and PKCα, to label rod bipolar cells. PKCαimmunopositive cells did not contain DSCAM. K and L, Sections of DscamFD/FD retina were stained with antibodies to Calbindin to label horizontal cells (K) and a subset of amacrine cells (L) (N=4 retinas). K, Horizontal cells, identified by their location close to the outer plexiform layer, did not contain DSCAM. L, Dimly staining calbindin-positive (ChAT-negative) amacrine cells located within the inner nuclear layer contained DSCAM protein. M and N, Sections of DscamFD/FD retina were stained with antibodies to calsenilin to label type 4 cone bipolar cells (M) and a subset of amacrine cells (N) (N=4 retinas). M, DSCAM immunoreactivity was observed in calsenilin positive bipolar cells. N, DSCAM protein was observed in some calsenilin positive amacrine cells, identified by their location in the inner nuclear layer. O, Sections of DscamFD/FD retina were stained with antibodies to DSCAM and NK3R, to label type 1 and 2 cone bipolar cells. NK3Rimmunopositive cells did not contain DSCAM. P, Sections of DscamFD/FD retina were stained with antibodies to DSCAM and recoverin, to label type 2 cone bipolar cells. Recoverinimmunopositive cells did not contain DSCAM. The scale bar in (A) is equivalent to 5 μm in A, F, H and I. The scale bar in (B) is equivalent to 5 μm in B and C. The scale bar in (D) is equivalent to 5 μm in D. The scale bar in (E) is equivalent to 5 μm in E. The scale bar in (G) is equivalent to 5 μm in G, J and L. The scale bar in (M) is equivalent to 5 μm in M–P.

Article Snippet: 1:2,000 HCN4 Amino acids 119–155 of human HCN4 (HGHLHDSAEERRLIAEGDASPGEDRTPPGLAAEPERP) Alomone, Jerusalem, Israel.

Techniques: Staining

Journal: The Journal of comparative neurology

Article Title: DSCAM Localization and Function at the Mouse Cone Synapse

doi: 10.1002/cne.23552

Figure Lengend Snippet: Dscam expressing cell types in the inner nuclear layer

Article Snippet: 1:2,000 HCN4 Amino acids 119–155 of human HCN4 (HGHLHDSAEERRLIAEGDASPGEDRTPPGLAAEPERP) Alomone, Jerusalem, Israel.

Techniques: Expressing, Marker

Journal: The Journal of comparative neurology

Article Title: DSCAM Localization and Function at the Mouse Cone Synapse

doi: 10.1002/cne.23552

Figure Lengend Snippet: A–C, Sections of wild type adult (>P42) retina stained with antibodies to DSCAM and PKARIIβ, calsenilin, HCN4 or Syt2 and PNA (N=4 retinas). DSCAM immunoreactivity was observed at the ends of some cone bipolar cell dendrite tips that terminated at the cone pedicle. The percent of DSCAM puncta that were observed within the dendrites of the respective bipolar cell types was counted for twenty pedicles and is reported in the upper corner of the left most panel. The result of this count when the channel containing bipolar cell staining was flipped along the horizontal axis is reported in the lower left hand corner. The scale bar in (A) is equivalent to 5 μm.

Article Snippet: 1:2,000 HCN4 Amino acids 119–155 of human HCN4 (HGHLHDSAEERRLIAEGDASPGEDRTPPGLAAEPERP) Alomone, Jerusalem, Israel.

Techniques: Staining

Journal: The Journal of comparative neurology

Article Title: DSCAM Localization and Function at the Mouse Cone Synapse

doi: 10.1002/cne.23552

Figure Lengend Snippet: Primary Antibodies

Article Snippet: HCN4 , Amino acids 119–155 of human HCN4 (HGHLHDSAEERRLIAEGDASPGEDRTPPGLAAEPERP) , Alomone, Jerusalem, Israel. Catalog Number: APC-052. Rabbit Polyclonal. , 1:500.

Techniques: Recombinant, Purification, Transduction

Journal: The Journal of comparative neurology

Article Title: DSCAM Localization and Function at the Mouse Cone Synapse

doi: 10.1002/cne.23552

Figure Lengend Snippet: A–D, Sections of adult (>P42) wild type and Dscam2J/2J retina were stained with antibodies to NK3R to label type 1 and type 2 cone bipolar cells, and PNA (N=4 retinas). Differences when comparing wild type and Dscam loss of function retina were not detected. E–F, Sections of adult (>P42) wild type and Dscam2J/2J retina were stained with antibodies to recoverin to label type 2 cone bipolar cells and rods, and PNA (N=4 retinas). Differences when comparing wild type and Dscam loss of function retina were not detected. I–L, Sections of adult (>P42) wild type and Dscam2J/2J retina were stained with HCN4 to label type 3a cone bipolar cells and PNA (N=4 retinas). Lamination of HCN4 positive neurites in the ON half of the inner plexiform layer are longer in the Dscam2J/2J retina compared to wild type controls (I versus J; arrow). No differences were detected in the outer plexiform layer. M–T, Sections of adult (>P42) wild type and Dscam2J/2J retina were stained with antibodies to PKARIIβ to label type 3b cone bipolar cells and PNA (N>4 retinas). PKARIIβ immunopositive bipolar cells have clumped dendrite arbors proximal to cone pedicles, compared to wild type (P, S and T; mutant vs. O, Q and R; wild type). The scale bar in (A) is equivalent to 100 μm in A, B, E, F, I, J, M and N. The scale bar in (C) is equivalent to 20 μm in C, D, G, H, K, L, O and P. The scale bar in (Q) is equivalent to 10 μm in Q–T.

Article Snippet: HCN4 , Amino acids 119–155 of human HCN4 (HGHLHDSAEERRLIAEGDASPGEDRTPPGLAAEPERP) , Alomone, Jerusalem, Israel. Catalog Number: APC-052. Rabbit Polyclonal. , 1:500.

Techniques: Staining, Mutagenesis

Journal: The Journal of comparative neurology

Article Title: DSCAM Localization and Function at the Mouse Cone Synapse

doi: 10.1002/cne.23552

Figure Lengend Snippet: A, Section of DscamFD/FD retina stained with antibodies to DSCAM and Brn3A (N=2). DSCAM immunoreactivity was observed in the soma of Brn3A positive cells in the retinal ganglion cell layer. B, Section of DscamFD/FD retina stained with antibodies to DSCAM and disabled (Dab) to stain AII amacrine cells (N=2). AII amacrine cells did not contain DSCAM in their soma. C, Section of DscamFD/FD retina stained with antibodies to DSCAM and ChAT, to label cholinergic amacrine cells (N=2). Cholinergic amacrine cell soma in the inner nuclear layer (top) or retinal ganglion layer (bottom) did not contain DSCAM protein. D, Section of DscamFD/FD retina stained with antibodies to DSCAM and bNOS (N=2). bNOS positive amacrine cells in both the inner nuclear layer (top) and retinal ganglion cell layer (bottom) contained DSCAM protein in their soma. E, Section of DscamFD/FD retina stained with antibodies to DSCAM and Chx10, to label bipolar cells (N=4). DSCAM immunoreactivity was observed in the soma surrounding some Chx10 positive cells. F and G, Section of DscamFD/FD retina stained with antibodies to HCN4 to label a subset of amacrine cells (G) and type 3a cone bipolar cells (F) (N=2 retinas). F, HCN4-positive bipolar cells, distinguished from amacrine cells by projection of dendrites to the outer plexiform layer and location, contained DSCAM protein in their soma. G, HCN4-positive amacrine cells, identified by their location proximal to the inner plexiform layer, did not contain DSCAM protein. H, Sections of DscamFD/FD retina were stained with antibodies to DSCAM and PKARIIβ to label type 3b bipolar cells. DSCAM immunoreactivity was observed in PKARIIβ positive cells. I, Sections of DscamFD/FD retina were stained with antibodies to DSCAM and Syt2, to label type 2 and type 6 bipolar cells (N=2 retinas). Some Syt2 immunopositive cells contained DSCAM immunoreactivity while others did not. J, Sections of DscamFD/FD retina were stained with antibodies to DSCAM and PKCα, to label rod bipolar cells. PKCαimmunopositive cells did not contain DSCAM. K and L, Sections of DscamFD/FD retina were stained with antibodies to Calbindin to label horizontal cells (K) and a subset of amacrine cells (L) (N=4 retinas). K, Horizontal cells, identified by their location close to the outer plexiform layer, did not contain DSCAM. L, Dimly staining calbindin-positive (ChAT-negative) amacrine cells located within the inner nuclear layer contained DSCAM protein. M and N, Sections of DscamFD/FD retina were stained with antibodies to calsenilin to label type 4 cone bipolar cells (M) and a subset of amacrine cells (N) (N=4 retinas). M, DSCAM immunoreactivity was observed in calsenilin positive bipolar cells. N, DSCAM protein was observed in some calsenilin positive amacrine cells, identified by their location in the inner nuclear layer. O, Sections of DscamFD/FD retina were stained with antibodies to DSCAM and NK3R, to label type 1 and 2 cone bipolar cells. NK3Rimmunopositive cells did not contain DSCAM. P, Sections of DscamFD/FD retina were stained with antibodies to DSCAM and recoverin, to label type 2 cone bipolar cells. Recoverinimmunopositive cells did not contain DSCAM. The scale bar in (A) is equivalent to 5 μm in A, F, H and I. The scale bar in (B) is equivalent to 5 μm in B and C. The scale bar in (D) is equivalent to 5 μm in D. The scale bar in (E) is equivalent to 5 μm in E. The scale bar in (G) is equivalent to 5 μm in G, J and L. The scale bar in (M) is equivalent to 5 μm in M–P.

Article Snippet: HCN4 , Amino acids 119–155 of human HCN4 (HGHLHDSAEERRLIAEGDASPGEDRTPPGLAAEPERP) , Alomone, Jerusalem, Israel. Catalog Number: APC-052. Rabbit Polyclonal. , 1:500.

Techniques: Staining

Journal: The Journal of comparative neurology

Article Title: DSCAM Localization and Function at the Mouse Cone Synapse

doi: 10.1002/cne.23552

Figure Lengend Snippet: Dscam expressing cell types in the inner nuclear layer

Article Snippet: HCN4 , Amino acids 119–155 of human HCN4 (HGHLHDSAEERRLIAEGDASPGEDRTPPGLAAEPERP) , Alomone, Jerusalem, Israel. Catalog Number: APC-052. Rabbit Polyclonal. , 1:500.

Techniques: Expressing, Marker

Journal: The Journal of comparative neurology

Article Title: DSCAM Localization and Function at the Mouse Cone Synapse

doi: 10.1002/cne.23552

Figure Lengend Snippet: A–C, Sections of wild type adult (>P42) retina stained with antibodies to DSCAM and PKARIIβ, calsenilin, HCN4 or Syt2 and PNA (N=4 retinas). DSCAM immunoreactivity was observed at the ends of some cone bipolar cell dendrite tips that terminated at the cone pedicle. The percent of DSCAM puncta that were observed within the dendrites of the respective bipolar cell types was counted for twenty pedicles and is reported in the upper corner of the left most panel. The result of this count when the channel containing bipolar cell staining was flipped along the horizontal axis is reported in the lower left hand corner. The scale bar in (A) is equivalent to 5 μm.

Article Snippet: HCN4 , Amino acids 119–155 of human HCN4 (HGHLHDSAEERRLIAEGDASPGEDRTPPGLAAEPERP) , Alomone, Jerusalem, Israel. Catalog Number: APC-052. Rabbit Polyclonal. , 1:500.

Techniques: Staining

Journal: Cell and Tissue Research

Article Title: Hyperpolarization-activated and cyclic nucleotide-gated channels are differentially expressed in juxtaglomerular cells in the olfactory bulb of mice

doi: 10.1007/s00441-009-0904-9

Figure Lengend Snippet: Primary antibodies. If no source is stated, cyclic-nucleotide-gated ( HCN ) antibodies were generated as described in Materials and methods

Article Snippet: HCN4alo , Rabbit polyclonal , 1:500 , Western blot, immunohistochemistry; results identical to independent antibody , Human HCN4 aa119–155/anti-HCN4, Alomone, Jerusalem, Israel, prod. no. APC-052, LOT AN02 , Feigenspan et al. ( ) .

Techniques: Generated, Western Blot, Immunohistochemistry, Immunoprecipitation, Purification, Immunocytochemistry, Immunofluorescence, Sequencing, Transduction, RIA Assay, Recombinant

Journal: Cell and Tissue Research

Article Title: Hyperpolarization-activated and cyclic nucleotide-gated channels are differentially expressed in juxtaglomerular cells in the olfactory bulb of mice

doi: 10.1007/s00441-009-0904-9

Figure Lengend Snippet: Immunohistochemical fingerprints ( IF ) in juxtaglomerular cells. IF1–IF9 ( arrowheads ) are found in PG-like cells, IF10 and IF11 ( barred arrows ) are found in SA-like cells, and IF12–IF17 ( arrows ) are found in ET-like cells ( + to ++++ level of staining intensity, +/- weak staining detected in some cells of the respective cell population probably reflecting expression at the detection threshold, - no detected staining, M the indicated protein usually found within the cytosol but antibody staining appeared as plasma membrane staining in cells with IF1). Color and arrow code applies to figures, including those in the Electronic Supplementary Material

Article Snippet: HCN4alo , Rabbit polyclonal , 1:500 , Western blot, immunohistochemistry; results identical to independent antibody , Human HCN4 aa119–155/anti-HCN4, Alomone, Jerusalem, Israel, prod. no. APC-052, LOT AN02 , Feigenspan et al. ( ) .

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: Cell and Tissue Research

Article Title: Hyperpolarization-activated and cyclic nucleotide-gated channels are differentially expressed in juxtaglomerular cells in the olfactory bulb of mice

doi: 10.1007/s00441-009-0904-9

Figure Lengend Snippet: Survey of HCN channel expression in the olfactory bulb. a Antibodies against HCN1–HCN4 ( HCN1 RTQ-7C3, HCN2 HCN2α, HCN3 HCN3 Shigemoto, HCN4 HCN4γ) recognize the glycosylated and degylcosylated HCN isoforms in Western blots of proteins from mouse olfactory bulb. The enzyme PNGaseF was used to deglycosylate HCN channels ( + lanes ). Molecular weights of deglycosylated HCN channels were 102 kDa (HCN1), 94 kDa (HCN2), 86 kDa (HCN3), and 130 kDa (HCN4). Comparison of molecular weights with ( + lanes ) and without ( − lanes ) PNGaseF treatment shows that all HCN isoforms are glycosylated ( numbers right molecular weight markers in kDa). b , c Representative example of double-labeling with two antibodies ( b QQA-1A6, c HCN2α) that recognize different epitopes of one and the same HCN isoform (HCN2). The staining patterns produced by the two antibodies are identical. Field of view reaches from the glomerular layer ( top ) to the external plexiform layer ( bottom ). Bar 20 μm. d–g Immunohistochemical staining shows the distribution of HCN1 ( d ), HCN2 ( e ), HCN3 ( f ), and HCN4 ( e ) within the various layers of the olfactory bulb in 20-μm-thick coronal sections ( EPL external plexiform layer, GL glomerular layer, GrL granule cell layer, IPL internal plexiform layer, ML mitral cell layer, AL olfactory nerve layer). Antibodies used were: HCN1α ( d ), QQA-1A6 ( e ), HCN3α4 ( f ), PG2-1A4 ( g ). Bar 100 μm

Article Snippet: HCN4alo , Rabbit polyclonal , 1:500 , Western blot, immunohistochemistry; results identical to independent antibody , Human HCN4 aa119–155/anti-HCN4, Alomone, Jerusalem, Israel, prod. no. APC-052, LOT AN02 , Feigenspan et al. ( ) .

Techniques: Expressing, Western Blot, Molecular Weight, Labeling, Staining, Produced, Immunohistochemical staining

Journal: Cell and Tissue Research

Article Title: Hyperpolarization-activated and cyclic nucleotide-gated channels are differentially expressed in juxtaglomerular cells in the olfactory bulb of mice

doi: 10.1007/s00441-009-0904-9

Figure Lengend Snippet: Antibody staining in PG-like cell populations. a–d Example of marker staining ( green CR, red NOS, blue vilip1), which partially overlaps in PG-like cells ( arrowheads ). Antibodies used were: CAL, NOSmono, and vilip1. e–h Different co-localizations of HCN isoforms ( green HCN1, red HCN4) and marker staining ( blue CB) in PG-like cells ( arrowheads ). Antibodies used were: HCN1α, PG2-1A4, and CabPneu. Further staining is shown in Electronic Supplementary Material, Fig. . All staining patterns are summarized in Table . Color codes of arrowheads as in Table . Bars 10 μm

Article Snippet: HCN4alo , Rabbit polyclonal , 1:500 , Western blot, immunohistochemistry; results identical to independent antibody , Human HCN4 aa119–155/anti-HCN4, Alomone, Jerusalem, Israel, prod. no. APC-052, LOT AN02 , Feigenspan et al. ( ) .

Techniques: Staining, Marker

Journal: Cell and Tissue Research

Article Title: Hyperpolarization-activated and cyclic nucleotide-gated channels are differentially expressed in juxtaglomerular cells in the olfactory bulb of mice

doi: 10.1007/s00441-009-0904-9

Figure Lengend Snippet: Antibody staining in two SA-like cell populations. a–h Immunohistochemical identification of SA-like cells with IF10. Cells with IF10 ( green barred arrow ) are characterized by the co-localization of HCN1, HCN3, HCN4, and NOS. a A cell with IF10 is labeled by antibodies directed against NOS (NOSpoly). A long (>100 μm) dendrite is visible. b–e Triple-staining for HCN1 ( blue HCN1α), NOS ( green NOSpoly), and HCN4 ( red PG2-1A4). Cells with IF10 show strong expression of NOS, weaker expression of HCN4, and expression around the detection threshold of HCN1. f–h Double-staining for NOS ( green NOSpoly) and HCN3 ( red HCN3 Shigemoto). i–k Immunohistochemical identification of cells with IF11 ( blue barred arrow ). These cells express HCN2 ( red QQA-1A6). Projection of a confocal LSM section series reveals that dendrites of cells with IF11 do not reach into glomeruli. Glomeruli are visible because of the ramification of vilip1-stained PG-like cells ( green vilip1) within glomeruli. Color codes of barred arrows as in Table . Bars 10 μm ( e , h , k ), 50 μm ( a )

Article Snippet: HCN4alo , Rabbit polyclonal , 1:500 , Western blot, immunohistochemistry; results identical to independent antibody , Human HCN4 aa119–155/anti-HCN4, Alomone, Jerusalem, Israel, prod. no. APC-052, LOT AN02 , Feigenspan et al. ( ) .

Techniques: Staining, Immunohistochemical staining, Labeling, Expressing, Double Staining

Journal: Cell and Tissue Research

Article Title: Hyperpolarization-activated and cyclic nucleotide-gated channels are differentially expressed in juxtaglomerular cells in the olfactory bulb of mice

doi: 10.1007/s00441-009-0904-9

Figure Lengend Snippet: Localization pattern of HCN channel isoforms in the glomerular layer. a–d Low magnification of triple-labeling for HCN1 ( green , RTQ-7C3), HCN3 ( red , HCN3 Shigemoto), and HCN4 ( blue , PG2-1A4) in the external plexiform layer ( EPL ) and glomerular layer ( GL ). e–o Example of various populations of PG- and SA-like cells. Three cell populations express only a single HCN isoform ( overlays in h , k , o : green arrowhead HCN1, blue barred arrow HCN2, dark blue arrowhead HCN4). This is demonstrated with triple-labeling ( e–h ) for HCN1 ( green HCN1α), HCN2 ( blue HCN2α), and HCN4 ( red PG2-1A4), double-labeling ( i–k ) for HCN2 ( green QQA-1A6) and HCN3 ( red HCN3 Shigemoto), and triple-labeling ( l–o ) for HCN1 ( green HCN1α), HCN3 ( red HCN3α4), and HCN4 ( blue PG2-1A4). The cell population expressing only HCN4 is also visible in s ( dark blue arrowheads ). Co-localization of either HCN1 ( green HCN1α) and HCN3 ( red HCN3 Shigemoto) or HCN1 ( green HCN1α) and HCN4 ( blue PG2-1A4) in two additional cell populations ( orange arrowhead , red arrowhead , respectively) is shown in o . p–s Three ET-like cell populations with different repertoires of HCN isoforms ( white arrow HCN1, HCN3, and HCN4, orange arrow HCN1 and HCN3, purple arrow HCN3 and HCN4) can be detected in the triple-staining for HCN1 ( green HCN1α), HCN3 ( red HCN3 Shigemoto), and HCN4 ( blue PG2-1A4). PG-like cells expressing only HCN4 are indicated ( dark blue arrowheads ). From these stainings, it is not apparent which of the cell populations identified in later experiments (Figs. , , ) are stained. However, by using the results of all staining performed, one can unequivocally identify the cell types here. For convenience, cell populations in this and the following figures are labeled with the uniform color and arrow code shown in Table . Glomeruli are indicated by an asterisk and delineated by a dashed white line . Bars 50 μm ( d ), 10 μm ( h , k , o , s )

Article Snippet: HCN4alo , Rabbit polyclonal , 1:500 , Western blot, immunohistochemistry; results identical to independent antibody , Human HCN4 aa119–155/anti-HCN4, Alomone, Jerusalem, Israel, prod. no. APC-052, LOT AN02 , Feigenspan et al. ( ) .

Techniques: Labeling, Expressing, Staining

Journal: Cell and Tissue Research

Article Title: Hyperpolarization-activated and cyclic nucleotide-gated channels are differentially expressed in juxtaglomerular cells in the olfactory bulb of mice

doi: 10.1007/s00441-009-0904-9

Figure Lengend Snippet: Do our antibodies label all populations of juxtaglomerular cells? Practically all juxtaglomerular cells have been stained with at least one of the antibodies against ( a ) markers ( green TH, NOS, CB, vilip1, CR, GFAP) or ( b ) HCN channels ( red HCN1, HCN2, HCN4). c Nuclei are stained by TOPRO-3 ( blue ). d Overlay of a–c . Confocal LSM sections rarely show unstained cells ( arrows ). Note: most unstained cells are positioned at the borders of the GL. Antibodies used: TH, CabPneu, NOSpoly, vilip1, CAL, PG2-1A4, QQA-1A6, HCN1α, and GFAP ( EPL external plexiform layer, GL glomerular layer, AL olfactory nerve layer). Bar 10 μm

Article Snippet: HCN4alo , Rabbit polyclonal , 1:500 , Western blot, immunohistochemistry; results identical to independent antibody , Human HCN4 aa119–155/anti-HCN4, Alomone, Jerusalem, Israel, prod. no. APC-052, LOT AN02 , Feigenspan et al. ( ) .

Techniques: Staining