{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T13:26:25Z","timestamp":1791293185708,"version":"4.1.0"},"reference-count":28,"publisher":"F1000 Research Ltd","license":[{"start":{"date-parts":[[2013,10,4]],"date-time":"2013-10-04T00:00:00Z","timestamp":1380844800000},"content-version":"unspecified","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/3.0\/"}],"content-domain":{"domain":["f1000research.com"],"crossmark-restriction":false},"short-container-title":["F1000Res"],"abstract":"<ns4:p>\n                    Rett syndrome is a neurological disorder caused by mutations in the\n                    <ns4:italic>MECP2<\/ns4:italic>\n                    gene. \u00a0MeCP2 transcripts are alternatively spliced to generate two protein isoforms (MeCP2_e1 and MeCP2_e2) that differ at their N-termini. Whilst mRNAs for both forms are expressed ubiquitously, the one for\n                    <ns4:italic>MeCP2_e1<\/ns4:italic>\n                    is more abundant than for\n                    <ns4:italic>MeCP2_e2<\/ns4:italic>\n                    in the central nervous system. In transfected cells, both protein isoforms are nuclear and colocalize with densely methylated heterochromatic foci. With a view to understanding the physiological contribution of each isoform, and their respective roles in the pathogenesis of Rett syndrome, we set out to generate isoform-specific anti-MeCP2 antibodies. To this end, we immunized rabbits against the peptides corresponding to the short amino-terminal portions that are different between the two isoforms. The polyclonal antibodies thus obtained specifically detected their respective isoforms of MeCP2 in Neuro2a (N2A) cells transfected to express either form. Both antisera showed comparable sensitivities when used for Western blot or immunofluorescence, and were highly specific for their respective isoform. When those antibodies were used on mouse tissues, specific signals were easily detected for Mecp2_e1, whilst Mecp2_e2 was very difficult to detect by Western blot, and even more so by immunofluorescence. Our results thus suggest that brain cells express low amounts of the Mecp2-e2 isoform. Our findings are compatible with recent reports showing that MeCP2_e2 is dispensable for healthy brain function, and that it may be involved in the regulation of neuronal apoptosis and embryonic development.\n                  <\/ns4:p>","DOI":"10.12688\/f1000research.2-204.v1","type":"journal-article","created":{"date-parts":[[2013,10,4]],"date-time":"2013-10-04T09:14:00Z","timestamp":1380878040000},"page":"204","update-policy":"https:\/\/doi.org\/10.12688\/f1000research.crossmark-policy","source":"Crossref","is-referenced-by-count":11,"title":["Isoform-specific anti-MeCP2 antibodies confirm that expression of the e1 isoform strongly predominates in the brain"],"prefix":"10.12688","volume":"2","author":[{"given":"Lara","family":"Kaddoum","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicolas","family":"Panayotis","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Honor\u00e9","family":"Mazarguil","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giuseppina","family":"Giglia-Mari","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9721-8844","authenticated-orcid":false,"given":"Jean Christophe","family":"Roux","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7264-2681","authenticated-orcid":false,"given":"Etienne","family":"Joly","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"2560","published-online":{"date-parts":[[2013,10,4]]},"reference":[{"key":"ref-1","doi-asserted-by":"publisher","first-page":"422-437","DOI":"10.1016\/j.neuron.2007.10.001","article-title":"The story of Rett syndrome: from clinic to neurobiology.","volume":"56","author":"M Chahrour","year":"2007","journal-title":"Neuron."},{"key":"ref-2","doi-asserted-by":"publisher","first-page":"185-188","DOI":"10.1038\/13810","article-title":"Rett syndrome is caused by mutations in X-linked MECP2, encoding methyl-CpG-binding protein 2.","volume":"23","author":"R Amir","year":"1999","journal-title":"Nat 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Genet."},{"key":"ref-6","doi-asserted-by":"publisher","first-page":"306-321","DOI":"10.1016\/j.mcn.2004.07.006","article-title":"MECP2 is progressively expressed in post-migratory neurons and is involved in neuronal maturation rather than cell fate decisions.","volume":"27","author":"N Kishi","year":"2004","journal-title":"Mol Cell Neurosci."},{"key":"ref-7","doi-asserted-by":"publisher","first-page":"457-468","DOI":"10.1016\/j.molcel.2010.01.030","article-title":"Neuronal MeCP2 is expressed at near histone-octamer levels and globally alters the chromatin state.","volume":"37","author":"P Skene","year":"2010","journal-title":"Mol Cell."},{"key":"ref-8","doi-asserted-by":"publisher","first-page":"415-426","DOI":"10.1038\/nrg1878","article-title":"Molecular genetics of Rett syndrome: when DNA methylation goes unrecognized.","volume":"7","author":"T Bienvenu","year":"2006","journal-title":"Nat Rev Genet."},{"key":"ref-9","doi-asserted-by":"publisher","first-page":"339-341","DOI":"10.1038\/ng1327","article-title":"A previously unidentified MECP2 open reading frame defines a new protein isoform relevant to Rett syndrome.","volume":"36","author":"G Mnatzakanian","year":"2004","journal-title":"Nat Genet."},{"key":"ref-10","doi-asserted-by":"publisher","first-page":"1818-1823","DOI":"10.1093\/nar\/gkh349","article-title":"The major form of MeCP2 has a novel N-terminus generated by alternative splicing.","volume":"32","author":"S Kriaucionis","year":"2004","journal-title":"Nucleic Acids Res."},{"key":"ref-11","doi-asserted-by":"publisher","first-page":"526-542","DOI":"10.1002\/cne.21264","article-title":"Differential distribution of the MeCP2 splice variants in the postnatal mouse brain.","volume":"501","author":"J Dragich","year":"2007","journal-title":"J Comp Neurol."},{"key":"ref-12","doi-asserted-by":"publisher","first-page":"1019-1023","DOI":"10.1002\/ajmg.a.32776","article-title":"Novel exon 1 mutations in MECP2 implicate isoform MeCP2_e1 in classical Rett syndrome.","volume":"149A","author":"C Saunders","year":"2009","journal-title":"Am J Med Genet A."},{"key":"ref-13","article-title":"Isoform-selective MeCP2-e1 deficient mice recapitulate the neurologic defecits of Rett syndrome.","author":"D Yasui","year":"2012"},{"key":"ref-14","doi-asserted-by":"publisher","first-page":"210-216","DOI":"10.1002\/ajmg.b.32015","article-title":"Mutations in MECP2 exon 1 in classical Rett patients disrupt MECP2_e1 transcription, but not transcription of MECP2_e2.","volume":"159B","author":"P Gianakopoulos","year":"2012","journal-title":"Am J Med Genet B Neuropsychiatr Genet."},{"key":"ref-15","doi-asserted-by":"publisher","first-page":"13859-13867","DOI":"10.1074\/jbc.M111.309864","article-title":"Methyl CpG-binding protein isoform MeCP2_e2 is 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Neurosci."},{"key":"ref-28","doi-asserted-by":"publisher","first-page":"e49763","DOI":"10.1371\/journal.pone.0049763","article-title":"Novel MeCP2 isoform-specific antibody reveals the endogenous MeCP2E1 expression in murine brain, primary neurons and astrocytes.","volume":"7","author":"R Zachariah","year":"2012","journal-title":"PLoS 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status","group":{"name":"current-referee-status","label":"Current Referee Status"}},{"value":"10.5256\/f1000research.2104.r2161, Mario Costa, Institute of Neuroscience, National Research Council, Italy, Pisa, Italy, 23 Oct 2013, version 1, indexed","URL":"https:\/\/f1000research.com\/articles\/2-204\/v1#referee-response-2161","order":0,"name":"referee-response-2161","label":"Referee Report","group":{"name":"article-reports","label":"Article Reports"}},{"value":"10.5256\/f1000research.2104.r2346, Jean-Fran\u00e7ois Brunet, D\u00e9partement de Biologie, Institut de Biologie de l'Ecole normale sup\u00e9rieure, Paris, 75005, France, 13 Nov 2013, version 1, indexed","URL":"https:\/\/f1000research.com\/articles\/2-204\/v1#referee-response-2346","order":1,"name":"referee-response-2346","label":"Referee Report","group":{"name":"article-reports","label":"Article Reports"}},{"value":"10.5256\/f1000research.2104.r2439, Mike Johnston, Division of Neurology & Developmental Medicine, Kennedy Krieger Institute, Baltimore, USA, 14 Nov 2013, version 1, indexed","URL":"https:\/\/f1000research.com\/articles\/2-204\/v1#referee-response-2439","order":2,"name":"referee-response-2439","label":"Referee Report","group":{"name":"article-reports","label":"Article Reports"}},{"value":"<b>Anita Bandrowski<\/b>; \n<i>Posted: 23 Feb 2016<\/i>; Dear Authors,In testing a new tool for methods review, I detected the following research resources in your paper which did not contain the RRID (\n<u>R<\/u>esearch&nbsp;\n<u>R<\/u>esource&nbsp;\n<u>ID<\/u>entifier). As discussed in the instruction to authors, please consider adding these to the final version of your paper.RRIDs are universal and persistent product codes and are used for antibodies, software tools and model organisms, see instructions to authors:&nbsp;http:\/\/f1000research.com\/for-authors\/article-guidelines\/research-notesPlease consider verifying the accuracy of the RRID suggestions below:\n<i><b>Needs attention, not an exact match:<\/b><\/i>\n<b>Abcam Cat# ab2828 RRID:AB_2143853<\/b>\n<i><b>&nbsp;&nbsp;&nbsp;&nbsp; &nbsp;verify:&nbsp;<\/b><\/i>scicrunch.org\/resolver\/RRID:AB_2143853&nbsp;\n<i><b>&nbsp;&nbsp; &nbsp;or search:&nbsp;<\/b><\/i>scicrunch.org\/resources\/Any\/search?q=2828\n<i><b>NO MATCH<\/b><\/i>\n<i><b>B6.129P2(C)-Mecp2tm1.1Bird\/J &nbsp;<\/b><\/i>&nbsp; &nbsp; \n<i><b>search: <\/b><\/i>https:\/\/scicrunch.org\/resources\/Any\/search?q=mecp2%20Mouse%20JAX\n<b>Match found<\/b>\n<b>ImageJ, <\/b>Version 1.47&nbsp;\n<b>RRID:SCR_003070<\/b>\n<i><b>&nbsp; &nbsp; verify:&nbsp;<\/b><\/i>scicrunch.org\/resolver\/RRID:SCR_003070&nbsp;In the text several other antibodies were detected with no RRID. Please consider registering these via:antibodyregistry.ord\/addPlease consider verifying each antibody highlighted above against the identifier and add the syntax (vendor catalog RRID) to the final version of the paper.Kind regards,Anita","URL":"https:\/\/f1000research.com\/articles\/2-204\/v1#article-response-1829","order":3,"name":"article-response-1829","label":"Article Comment","group":{"name":"article-comments","label":"Article Comments"}},{"value":"This work was supported by Association Fran\u00e7aise du Syndrome de Rett (AFSR), the Fondation pour la Recherche Medicale (FRM) and the E-RARE EuroRETT network. JCR and EJ were supported by INSERM (Institut National pour la Sant\u00e9 et la Recherche M\u00e9dicale). NP was supported by an INSERM-R\u00e9gion Provence-Alpes-C\u00f4te d\u2019Azur fellowship, HM was supported by CNRS (Centre National pour la Recherche Scientifique).","order":4,"name":"grant-information","label":"Grant Information"},{"value":"This is an open access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.Data associated with the article are available under the terms of the Creative Commons Zero \"No rights reserved\" data waiver (CC0 1.0 Public domain dedication).","order":0,"name":"copyright-info","label":"Copyright"}]}}