rabbit polyclonal anti-dream sc-9142  (Santa Cruz Biotechnology)

Journal: International Journal of Molecular Sciences

Article Title: Cellular and Subcellular Localisation of Kv4-Associated KChIP Proteins in the Rat Cerebellum

doi: 10.3390/ijms21176403

Figure Lengend Snippet: Regional and cellular distribution of Kv channel interacting proteins (KChIPs) subunits KChIP3 and KChIP4 in the cerebellum. ( A – D ) Immunoreactivity for KChIP3 and KChIP4 in the cerebellar cortex using a pre-embedding immunoperoxidase method at the light microscopic level. Parasagittal photomicrographs of the cerebellar cortex. Immunoreactivity for both KChIP3 and KChIP4 was widely distributed in the cerebellar cortex with mostly overlapping labelling patterns. Although with some differences in intensity of labelling, strong immunoreactivity for KChIP3 and KChIP4 was found in the granule cell layer (gcl) and weaker in the molecular layer (ml) and the white matter (wm) was devoid of any staining. In the molecular layer, KChIP3 and KChIP4 was mostly neuropilar and KChIP3 labelling was also detected in cell bodies and dendrites of stellate and basket cells (black arrows). In the granule cell layer, KChIP3 and KChIP4 particularly concentrated in glomeruli (white arrows) and surrounding GCs. Scale bars: ( A ), 50 µm; ( B ), 100 µm; ( C , D ), 25 µm.

Article Snippet: Other primary antibodies used were the following: (1) rabbit anti-Kv4.2 polyclonal (APC-023; Alomone Labs., Jerusalem, Israel), (2) rabbit anti-Kv4.3 polyclonal (APC-017; Alomone Labs., Israel) and (3) rabbit anti-KChIP3 polyclonal (Rb-Af400; aa.

Techniques: Staining

Journal: International Journal of Molecular Sciences

Article Title: Cellular and Subcellular Localisation of Kv4-Associated KChIP Proteins in the Rat Cerebellum

doi: 10.3390/ijms21176403

Figure Lengend Snippet: Subcellular localisation of Kv channel interacting protein (KChIP) subunit KChIP3 in the cerebellum. Electron micrographs showing immunoparticles for KChIP3 in the cerebellar cortex, as detected using the pre-embedding immunogold technique. ( A – D ) In dendritic shafts (Den) and spines (s) of PCs, located in the molecular layer, KChIP3 immunoparticles were observed postsynaptically along the extrasynaptic plasma membrane (arrows). Less frequently, KChIP3 immunoparticles were observed intracellularly (crossed arrows). Presynaptically, KChIP3 immunoparticles were detected along the extrasynaptic membrane and at intracellular sites (arrowheads) of parallel fibres (pf). ( E – H ) In the granule cell layer, KChIP3 immunoparticles were observed along the somatic plasma membrane (arrows) and dendrites (Den, arrows) and intracellular sites (crossed arrows) of granule cells (GC). Presynaptically, KChIP3 immunoparticles were observed along the plasma membrane (arrowheads) of mossy fibre axon terminals (mf), but most of them were localised intracellularly (double arrowheads) associated with cytoplasmic membranes. Scale bars: ( A – C ), 200 nm; ( D – H ), 250 nm.

Article Snippet: Other primary antibodies used were the following: (1) rabbit anti-Kv4.2 polyclonal (APC-023; Alomone Labs., Jerusalem, Israel), (2) rabbit anti-Kv4.3 polyclonal (APC-017; Alomone Labs., Israel) and (3) rabbit anti-KChIP3 polyclonal (Rb-Af400; aa.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Cellular and Subcellular Localisation of Kv4-Associated KChIP Proteins in the Rat Cerebellum

doi: 10.3390/ijms21176403

Figure Lengend Snippet: Compartmentalisation of Kv channel interacting protein (KChIP) subunit KChIP3 in cerebellar cells. ( A ) Bar graphs showing the percentage of immunoparticles for KChIP3 at post- and pre-synaptic compartments in the molecular layer. A total of 1487 immunoparticles in the molecular layer were analysed, of which 38.1% were postsynaptic and 61.9% were presynaptic. Postsynaptically, immunoparticles were detected in dendritic spines (21.7%) and in dendritic shafts (78.35%), distributed along the plasma membrane (49.6% in spines; 27.9% in dendrites) and at cytoplasmic sites (50.4% in spines; 72.1% in dendrites). Presynaptically, immunoparticles were detected in parallel fibres of ML (61.9%) distributed along the plasma membrane (25.5%) and at cytoplasmatic sites (74.5%). ( B ) Bar graphs showing the percentage of KChIP3 immunoparticles at post- and pre-synaptic compartments, and along the plasma membrane and intracellular sites in dendritic shafts of granule cells and mossy fibres in the granule cell layer. A total of 1087 immunoparticles in the granular cell layer were analysed, of which 63.5% were postsynaptic and 36.5% were presynaptic. Postsynaptically, immunoparticles were detected in dendrites of GCs (63.5%), distributed along the plasma membrane (27%) and at cytoplasmic sites (73%). Presynaptically, immunoparticles were detected in mossy fibre terminals (36.5%), distributed mostly at cytoplasmic sites (94%) and very few along the plasma membrane (6%). ( C ) Histogram showing the radial distribution of KChIP3 immunoparticles from the plasma membrane towards cytoplasmic sites in dendrites of GCs and mossy fibres. In dendrites, immunoparticles for KChIP3 showed a skewed frequency distribution in the plasma membrane direction, but in mossy fibres they were more equally distributed across the axoplasm. ( D ) Histogram showing the distribution of immunoreactive KChIP3 in relation to glutamate release sites in dendritic spines PCs. The data show the proportion of KChIP3 immunoparticles at a given distance from the edge of the postsynaptic density. About 68.9% of immunolabelled KChIP3 are located in a 60–300 nm wide band, and then the density decreased markedly further in the spine membrane.

Article Snippet: Other primary antibodies used were the following: (1) rabbit anti-Kv4.2 polyclonal (APC-023; Alomone Labs., Jerusalem, Israel), (2) rabbit anti-Kv4.3 polyclonal (APC-017; Alomone Labs., Israel) and (3) rabbit anti-KChIP3 polyclonal (Rb-Af400; aa.

Techniques: