Confocal analysis of 0.2 mm optical sections (n = 104 cells). |
To distinguish MDMs (RFP+) from MiDMs (GFP+). |
MDMs and MiDMs can be distinguished by cell volume and primary processes. |
SBF-SEM inspection in 0.2 mm sections from 14 lesions, 7 mice at EAE onset. |
To detect MDMs and MiDMs in SBF-SEM using cell volume and process criteria. |
Using criteria detected in the previous step, it is possible to distinguish MDMs and MiDMs in SBF-SEM images. |
SBF-SEM inspection of ultrastructure of MDMs and MiDMs. |
To detect ultrastructural characteristics of MDMs and MiDMs. |
MDMs and MiDMs show characteristic ultrastructural differences regarding their mitochondria, nuclei, osmiophilic granules and microvilli. |
Quantification of relation of MDMs (n = 169) and MiDMs (n = 86) to axoglial units (n = 29 intact axons, 46 demyelinated axons). |
To determine relationship of MDMs and MiDMs to axoglial units and characterize presence of myelin debris. |
Most (55/75; 73%) axoglial units are contacted in limited fashion by MDMs and MiDMs. If one cell type is present (20/75 cells), it’s nearly always (18/20 segments) MDMs. |
Reconstruction of 3D shape of four representative MDMs at axoglial units. |
To detect relationship of MDMs with axoglial units at EAE onset. |
In all, 49 MDMs interacting with axoglial units in absence of nearby MiDMs, 2-3 MDMs were attached to each (n = 18) axoglial unit. MDMs have close relationship with nodes of Ranvier (7 MDMs /75 axoglial units). 3D reconstructions showed four representative MDMs at axoglial units: show one carrying out active demyelination, three at nodes of Ranvier. |