Journal: Cell Reports

Article Title: Oligodendrocyte-derived LGI3 and its receptor ADAM23 organize juxtaparanodal Kv1 channel clustering for short-term synaptic plasticity

doi: 10.1016/j.celrep.2023.113634

Figure Lengend Snippet: ID-related LGI3 is uniquely expressed in oligodendrocytes (A) Domain organization of human LGI3. SP, signal peptide; LRR, leucine-rich repeat domain; EPTP, epitempin domain; N- and C-cap, N- and C-terminal cysteine caps in LRR. (B) Mapping of the ID variant on the modeled LGI3 structure. Left, Asp331 is mapped as red spheres. Right, close-up view of the Ca 2+ -binding site in the LGI3 EPTP β-propeller. Ca 2+ and water molecules are shown as gray and red spheres, respectively. Sticks, Ca 2+ -interacting residues; dotted lines, hydrogen bonds. (C) Shown are western blots (WBs) of LGIs-V5 in the conditioned medium (secreted) and the cell lysates. Bottom, quantification of the amounts of secreted LGI proteins. n = 3 experiments. Mann-Whitney U test. ∗ p < 0.05. Mean ± SD. (D) Fluorescence in situ hybridization (FISH) analysis for Lgi3 , Lgi1 , and Lgi2 expression in the mouse brain (P125). Red, Lgi3 , Lgi1 , or Lgi2 ; green, Mbp . White arrows, Mbp + / Lgi3 + cells; yellow arrows, Mbp – / Lgi3 + cells; white arrowheads, Mbp + / Lgi1 – or Mbp + / Lgi2 – cells; yellow arrowheads, Mbp – / Lgi1 + or Mbp – / Lgi2 + cells. Nuclear DNA was stained by DAPI (blue). CC, corpus callosum; Cb, cerebellum; WM, white matter; L5/6, cortical layers V and VI. Scale bars, 10 μm. (E) WB analysis of LGI family proteins in the lysates of the mouse brains and primary cultured cortical neurons. N-cadherin, a loading control. P, postnatal days; DIV, days in vitro . (F) WB analysis of Lgi3 LoxP/ΔE1 and Lgi3 LoxP/ΔE1 ; Mbp -Cre mouse brain lysates (P35). See also Figure S1 and Table S1 .

Article Snippet: Mouse monoclonal anti-N-Cadherin (Clone 32) , BD Biosciences , Cat#610920; RRID: AB_2077527.

Techniques: Variant Assay, Binding Assay, Western Blot, MANN-WHITNEY, Fluorescence, In Situ Hybridization, Expressing, Staining, Cell Culture, In Vitro

Journal: Cell Reports

Article Title: Oligodendrocyte-derived LGI3 and its receptor ADAM23 organize juxtaparanodal Kv1 channel clustering for short-term synaptic plasticity

doi: 10.1016/j.celrep.2023.113634

Figure Lengend Snippet:

Article Snippet: Mouse monoclonal anti-N-Cadherin (Clone 32) , BD Biosciences , Cat#610920; RRID: AB_2077527.

Techniques: Recombinant, Virus, Sequencing, Modification, Multiplex Assay, Positive Control, Negative Control, Mass Spectrometry, Knock-In, Knock-Out, Transgenic Assay, Software

Journal: Journal of Virology

Article Title: Hamster-Adapted Sin Nombre Virus Causes Disseminated Infection and Efficiently Replicates in Pulmonary Endothelial Cells without Signs of Disease

doi: 10.1128/JVI.03291-12

Figure Lengend Snippet: Serially passaging Sin Nombre virus results in increased viral replication in hamster lungs without histological changes associated with HPS. Histological analysis was carried out on lung samples from hamsters infected with passaged Sin Nombre virus and compared to time-matched samples from Andes virus-infected animals. In the initial passage (p1) of Sin Nombre virus, nucleoprotein antigen and RNA were undetectable by immunohistochemistry and in situ hybridization. However, by p5 and continuing through p20, viral antigen and RNA were readily detectable in pulmonary endothelial cells from hamsters infected with the passaged virus, with a distribution and intensity similar to that observed in Andes-infected hamsters. Despite increased replication, no histological lesions were noted in the lungs of hamsters infected with the passaged virus. By comparison, time-matched samples collected from Andes-infected hamsters demonstrated perivascular edema, which ultimately leads to diffuse pulmonary edema and death.

Article Snippet: To confirm the presence of virus in endothelial cells, lung specimens were costained with monoclonal antibodies targeting the nucleoprotein (anti-SNV NP clone 5F1/F7 and anti-ANDV NP clone 1A8/F6, 1:1,000 dilution each; Austral Biologicals) and polyclonal rabbit anti-CD31 antibodies (ab28364, 1:50 dilution; Abcam) with Alexa Fluor 594 (anti-mouse) and 488 (anti-rabbit) secondary antibodies (A11005 and {"type":"entrez-nucleotide","attrs":{"text":"A11034","term_id":"489250","term_text":"A11034"}} A11034 , respectively, each at a 1:200 dilution; Invitrogen).

Techniques: Passaging, Virus, Infection, Immunohistochemistry, In Situ Hybridization, Comparison

Journal: Journal of Virology

Article Title: Hamster-Adapted Sin Nombre Virus Causes Disseminated Infection and Efficiently Replicates in Pulmonary Endothelial Cells without Signs of Disease

doi: 10.1128/JVI.03291-12

Figure Lengend Snippet: Immunofluorescence staining for endothelial cell markers and hantaviral antigen in hamster lungs. Lung samples collected 9 days postinfection were costained for endothelial cell markers (CD31, green) and hantavirus nucleoprotein antigen (red). Lung specimens from hamsters infected with a hamster-adapted (passage 5) Sin Nombre virus (A) and Andes virus (B) exhibited the presence of hantaviral antigen in cells positive for CD31, supporting the notion that both viruses replicate in pulmonary endothelial cells.

Article Snippet: To confirm the presence of virus in endothelial cells, lung specimens were costained with monoclonal antibodies targeting the nucleoprotein (anti-SNV NP clone 5F1/F7 and anti-ANDV NP clone 1A8/F6, 1:1,000 dilution each; Austral Biologicals) and polyclonal rabbit anti-CD31 antibodies (ab28364, 1:50 dilution; Abcam) with Alexa Fluor 594 (anti-mouse) and 488 (anti-rabbit) secondary antibodies (A11005 and {"type":"entrez-nucleotide","attrs":{"text":"A11034","term_id":"489250","term_text":"A11034"}} A11034 , respectively, each at a 1:200 dilution; Invitrogen).

Techniques: Immunofluorescence, Staining, Infection, Virus