{"id":121838,"date":"2021-04-22T21:16:09","date_gmt":"2021-04-23T04:16:09","guid":{"rendered":"https:\/\/lifeboat.com\/blog\/2021\/04\/basp1-labels-neural-stem-cells-in-the-neurogenic-niches-of-mammalian-brain"},"modified":"2021-04-22T21:16:09","modified_gmt":"2021-04-23T04:16:09","slug":"basp1-labels-neural-stem-cells-in-the-neurogenic-niches-of-mammalian-brain","status":"publish","type":"post","link":"https:\/\/lifeboat.com\/blog\/2021\/04\/basp1-labels-neural-stem-cells-in-the-neurogenic-niches-of-mammalian-brain","title":{"rendered":"BASP1 labels neural stem cells in the neurogenic niches of mammalian brain"},"content":{"rendered":"<p style=\"padding-right: 20px\"><a class=\"aligncenter blog-photo\" href=\"https:\/\/lifeboat.com\/blog.images\/basp1-labels-neural-stem-cells-in-the-neurogenic-niches-of-mammalian-brain.jpg\"><\/a><\/p>\n<p>In this study we aimed to generate mouse antibodies against epitopes found on NPCs. We isolated one antibody (NSC-6) and characterized it in detail. Mass spectrometry using human hippocampal tissue revealed the identity of the recognized antigen as BASP1, a signaling protein that plays a key role in neurite outgrowth and plasticity<sup><a data-track=\"click\" data-track-action=\"reference anchor\" data-track-label=\"link\" data-test=\"citation-ref\" title=\"Wiederkehr, A., Staple, J. & Caroni, P. The motility-associated proteins GAP-43, MARCKS, and CAP-23 share unique targeting and surface activity-inducing properties. Exp. Cell Res. 236103&ndash;116 (1997).\" href=\"https:\/\/www.nature.com\/articles\/s41598-021-85129-1#ref-CR14\" id=\"ref-link-section-d11810e1599\">14<\/a>,<a data-track=\"click\" data-track-action=\"reference anchor\" data-track-label=\"link\" data-test=\"citation-ref\" title=\"Maekawa, S. et al. Cholesterol-dependent localization of NAP-22 on a neuronal membrane microdomain (raft). J. Biol. Chem. 274, 21369&ndash;21374 (1999).\" href=\"https:\/\/www.nature.com\/articles\/s41598-021-85129-1#ref-CR15\" id=\"ref-link-section-d11810e1599_1\">15<\/a>,<a data-track=\"click\" data-track-action=\"reference anchor\" data-track-label=\"link\" data-test=\"citation-ref\" title=\"Laux, T. et al. GAP43, MARCKS, and CAP23 modulate PI (4, 5) P  at plasmalemmal rafts, and regulate cell cortex actin dynamics through a common mechanism. J. Cell Biol. 149, 1455&ndash;1472 (2000).\" href=\"https:\/\/www.nature.com\/articles\/s41598-021-85129-1#ref-CR16\" id=\"ref-link-section-d11810e1599_2\">16<\/a>,<a data-track=\"click\" data-track-action=\"reference anchor\" data-track-label=\"link\" data-test=\"citation-ref\" title=\"Caroni, P., Aigner, L. & Schneider, C. Intrinsic neuronal determinants locally regulate extrasynaptic and synaptic growth at the adult neuromuscular junction. J. Cell Biol. 136679&ndash;692 (1997).\" href=\"https:\/\/www.nature.com\/articles\/s41598-021-85129-1#ref-CR17\" id=\"ref-link-section-d11810e1599_3\">17<\/a>,<a data-track=\"click\" data-track-action=\"reference anchor\" data-track-label=\"link\" data-test=\"citation-ref\" title=\"Frey, D., Laux, T., Xu, L., Schneider, C. & Caroni, P. Shared and unique roles of CAP23 and GAP43 in actin regulation, neurite outgrowth, and anatomical plasticity. J. Cell Biol. 149, 1443&ndash;1454 (2000).\" href=\"https:\/\/www.nature.com\/articles\/s41598-021-85129-1#ref-CR18\" id=\"ref-link-section-d11810e1599_4\">18<\/a>,<a data-track=\"click\" data-track-action=\"reference anchor\" data-track-label=\"link\" data-test=\"citation-ref\" aria-label=\"Reference 19\" title=\"Bomze, H. M., Bulsara, K. R., Iskandar, B. J., Caroni, P. & Skene, J. H. Spinal axon regeneration evoked by replacing two growth cone proteins in adult neurons. Nat. Neurosci. 4, 38&ndash;43 (2001).\" href=\"https:\/\/www.nature.com\/articles\/s41598-021-85129-1#ref-CR19\" id=\"ref-link-section-d11810e1602\">19<\/a><\/sup>, but here, we demonstrate that it might be utilized as a marker of NSCs in the adult brain.<\/p>\n<p>Similar approaches to developing antibodies against mouse embryonic stem cells have been attempted in the past utilizing mice<sup><a data-track=\"click\" data-track-action=\"reference anchor\" data-track-label=\"link\" data-test=\"citation-ref\" aria-label=\"Reference 46\" title=\"Choo, A. B. et al. Selection against undifferentiated human embryonic stem cells by a cytotoxic antibody recognizing podocalyxin-like protein-1. Stem Cells 26, 1454&ndash;1463 (2008).\" href=\"https:\/\/www.nature.com\/articles\/s41598-021-85129-1#ref-CR46\" id=\"ref-link-section-d11810e1609\">46<\/a>,<a data-track=\"click\" data-track-action=\"reference anchor\" data-track-label=\"link\" data-test=\"citation-ref\" aria-label=\"Reference 47\" title=\"Son, Y. S. et al. Heat shock 70-kDa protein 8 isoform 1 is expressed on the surface of human embryonic stem cells and downregulated upon differentiation. Stem Cells 23, 1502&ndash;1513 (2005).\" href=\"https:\/\/www.nature.com\/articles\/s41598-021-85129-1#ref-CR47\" id=\"ref-link-section-d11810e1612\">47<\/a><\/sup> and rabbits<sup><a data-track=\"click\" data-track-action=\"reference anchor\" data-track-label=\"link\" data-test=\"citation-ref\" aria-label=\"Reference 48\" title=\"Tan, Z. et al. Production of rabbit monoclonal antibodies against mouse embryonic stem cells and identification of pluripotency-associated surface antigens. J. Immunol. Methods 365149&ndash;157 (2011).\" href=\"https:\/\/www.nature.com\/articles\/s41598-021-85129-1#ref-CR48\" id=\"ref-link-section-d11810e1616\">48<\/a><\/sup>. Major drawbacks in mice include immune tolerance to mouse embryonic stem cell surface antigens leading to low antibody production, which could be overcome by immunizing rabbits instead. Regardless of the animal used as a host, a significant number of antibodies are typically generated against intracellular epitopes when animals are immunized with whole cells as was observed in our study.<\/p>\n<p>We found that NSC-6-labeled BASP1 localizes to all radial glia at the E12 stage of brain development, while postnatally, it restricts to the neurogenic areas of the mouse brain but not the cortex. This expression pattern contrasts previous study using DAB-based immunolabeling for <i>NAP-22<\/i> (BASP1 alias) in the adult rat brain, which demonstrated robust labeling of cerebral cortex<sup><a data-track=\"click\" data-track-action=\"reference anchor\" data-track-label=\"link\" data-test=\"citation-ref\" aria-label=\"Reference 27\" title=\"Iino, S., Kobayashi, S. & Maekawa, S. Immunohistochemical localization of a novel acidic calmodulin-binding protein, NAP-22, in the rat brain. Neuroscience 91, 1435&ndash;1444 (1999).\" href=\"https:\/\/www.nature.com\/articles\/s41598-021-85129-1#ref-CR27\" id=\"ref-link-section-d11810e1623\">27<\/a><\/sup>. While we do not know the basis of this difference in immunolabeling of cortex, possibilities include species variations between rat and mouse expression of BASP1, or differences in epitope recognition between the two antibodies used that could yield distinct patterns of immunoreactivity. Indeed, the two commercial BASP1 polyclonal antibodies did not immunolabel NSCs and in general, exhibited poor staining of the mouse brain tissue.<\/p>\n","protected":false},"excerpt":{"rendered":"<p>In this study we aimed to generate mouse antibodies against epitopes found on NPCs. We isolated one antibody (NSC-6) and characterized it in detail. Mass spectrometry using human hippocampal tissue revealed the identity of the recognized antigen as BASP1, a signaling protein that plays a key role in neurite outgrowth and plasticity14,15,16,17,18,19, but here, we [\u2026]<\/p>\n","protected":false},"author":427,"featured_media":0,"comment_status":"open","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[11,47],"tags":[],"class_list":["post-121838","post","type-post","status-publish","format-standard","hentry","category-biotech-medical","category-neuroscience"],"_links":{"self":[{"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/posts\/121838","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/users\/427"}],"replies":[{"embeddable":true,"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/comments?post=121838"}],"version-history":[{"count":0,"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/posts\/121838\/revisions"}],"wp:attachment":[{"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/media?parent=121838"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/categories?post=121838"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/tags?post=121838"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}