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Grant support

We thank Merche Rivas, Maria Marsal, Gustavo Castro, Jordi Cortes, Alina Hirschmann, and Angel Sandoval (ICFOInstitut de Ciencies Fotoniques) and Mercedes Alba, Marija Radosevic, David Soto, Xavier Gasull, Mar Guasp, and Lidia Sabater (IDIBAPS, Hospital Clinic, University of Barcelona) for their technical support and for providing of reagents, and Josep Dalmau and Myrna R. Rosenfeld (IDIBAPS, Hospital Clinic, University of Barcelona) for their critical review of the manuscript and mentorship. This study was funded by Instituto de Salud Carlos III (ISCIII) and co-financed by the European Union, FIS (PI20/00280, J.P.), Fundacio CELLEX (P.L-A.); Ministerio de Economia y Competitividad culturing and studying other developmental tissues at similar stages such as the notochord, neural plate and lateral plate mesoderm.

Analysis of institutional authors

Aguilar EAuthorPlanaguma JCorresponding Author
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Article

Hippocampal Neuronal Cultures to Detect and Study New Pathogenic Antibodies Involved in Autoimmune Encephalitis

Publicated to:Jove-Journal Of Visualized Experiments. (184): e63829- - 2022-06-01 (184), DOI: 10.3791/63829

Authors: Cunquero, Marina; Aguilar, Esther; Loza-Alvarez, Pablo; Planaguma, Jesus

Affiliations

Barcelona Inst Sci & Technol, ICFO Inst Ciencies Foton, Barcelona, Spain - Author
Univ Barcelona, Inst Invest Biomed August Pi & Sunyer IDIBAPS, Hosp Clin, Barcelona, Spain - Author

Abstract

Over the last 15 years, a new category of antibody-mediated diseases of the central nervous system (CNS) has been characterized and is now defined as "autoimmune encephalitis" (AE). There are currently 17 known AE syndromes, and all are associated with antibodies against the neuronal cell surface or synaptic proteins. The clinical syndromes are complex and vary according to the type of associated antibody. The best-known of these diseases is anti-N-methyl D-aspartate receptor (NMDAR) encephalitis, which is a prominent neuropsychiatric disorder associated with severe memory and behavioral impairments. The associated antibodies react with the GluN1 subunit of the NMDAR at the N-terminal domain. The approach most frequently used for the discovery and characterization of AE antibodies includes the culture of dissociated, fetal, rodent hippocampal neurons. During the process of antibody characterization, live neurons in culture are exposed to patients' serum or CSF, and the detection of reactivity indicates that the serum or CSF samples of the patient contain antibodies against neuronal surface antigens. Hippocampal cultures can also be used to determine whether the antibodies in patients are potentially pathogenic by examining if they cause structural or functional alterations of the neurons. The level of success of these studies depends on the quality of the cultures and on the protocols used to obtain and detect the reactivity of patient samples. This article provides an optimized protocol for primary cell culture of fetal rat hippocampal neurons combined with immunostaining to determine the presence of antibodies in the serum or CSF of patients. An example of how to examine the potential pathogenic effects of NMDAR antibodies using cultured neurons and calcium imaging is also presented.

Keywords
autoantibodiescase seriesmechanismsAnimalAnimalsAnti nmda receptor encephalitisAnti-n-methyl-d-aspartate receptor encephalitisAutoantibodiesAutoantibodyEncephalitisHashimoto diseaseHippocampusHumanHumansMetabolismNerve cellNeuronsRatRatsReceptor antibodiesSyndrome

Quality index

Bibliometric impact. Analysis of the contribution and dissemination channel

The work has been published in the journal Jove-Journal Of Visualized Experiments due to its progression and the good impact it has achieved in recent years, according to the agency Scopus (SJR), it has become a reference in its field. In the year of publication of the work, 2022, it was in position , thus managing to position itself as a Q2 (Segundo Cuartil), in the category Chemical Engineering (Miscellaneous). Notably, the journal is positioned en el Cuartil Q3 for the agency WoS (JCR) in the category Multidisciplinary Sciences.

Independientemente del impacto esperado determinado por el canal de difusión, es importante destacar el impacto real observado de la propia aportación.

Según las diferentes agencias de indexación, el número de citas acumuladas por esta publicación hasta la fecha 2025-05-23:

  • WoS: 2
  • Scopus: 2
Impact and social visibility

From the perspective of influence or social adoption, and based on metrics associated with mentions and interactions provided by agencies specializing in calculating the so-called "Alternative or Social Metrics," we can highlight as of 2025-05-23:

  • The use, from an academic perspective evidenced by the Altmetric agency indicator referring to aggregations made by the personal bibliographic manager Mendeley, gives us a total of: 10.
  • The use of this contribution in bookmarks, code forks, additions to favorite lists for recurrent reading, as well as general views, indicates that someone is using the publication as a basis for their current work. This may be a notable indicator of future more formal and academic citations. This claim is supported by the result of the "Capture" indicator, which yields a total of: 10 (PlumX).

With a more dissemination-oriented intent and targeting more general audiences, we can observe other more global scores such as:

  • The Total Score from Altmetric: 1.
  • The number of mentions on the social network X (formerly Twitter): 2 (Altmetric).

It is essential to present evidence supporting full alignment with institutional principles and guidelines on Open Science and the Conservation and Dissemination of Intellectual Heritage. A clear example of this is:

  • The work has been submitted to a journal whose editorial policy allows open Open Access publication.
Leadership analysis of institutional authors

There is a significant leadership presence as some of the institution’s authors appear as the first or last signer, detailed as follows: Last Author (Planaguma i Valls, Jesus).

the author responsible for correspondence tasks has been Planaguma i Valls, Jesus.