Object-based analysis of astroglial reaction and astrocyte subtype morphology after ischemic brain injury

• Autorzy: Wagner D.-C. , Scheibe J. , Glocke I. , Weise G. , Deten A. , Boltze J. , Kranz A.
• Języki publikacji: EN

Abstrakty

EN

The astrocytic response to ischemic brain injury is characterized by specific alterations of glial cell morphology and function. Various studies described both beneficial and detrimental aspects of activated astrocytes, suggesting the existence of different subtypes. We investigated this issue using a novel object-based approach to study characteristics of astrogliosis after stroke. Spontaneously hypertensive rats received permanent middle cerebral artery occlusion. After 96 h, brain specimens were removed, fixed and stained for GFAP, glutaminę synthetase (GS), S100P and Musashil (Mshl). Three regions of interest were defined (contralateral hemisphere, ipsilateral remote zone and infarct border zone), and confocal stacks were acquired (n=5 biological with each n=4 technical replicates). The stacks were background-corrected and colocalization between the selected markers and GFAP was determined using an automated thresholding algorithm. The fluorescence and colocalization channels were then converted into 3D-objects using both intensity and volume as filters to ultimately determine the final volumes of marker expression and colocalization, as well as the morphological changes of astrocyte process arborisation. We found that both S100P and Mshl determined the same GFAP-positive astroglial cell population albeit the cellular compartments differed. GFAP stained most of the astrocyte processes and is hence suitable for the analysis of qualitative characteristics of astrogliosis. Due to its peri-nuclear localization, Mshl is appropriate to estimate the total number of astrocytes even in regions with severe reactive astrogliosis. GS expression in GFAP-positive astrocytes was high in the remote zone and low at the infarct border, indicating the existence of astrocyte subclasses.

• Wydawca: -
• Czasopismo: Acta Neurobiologiae Experimentalis
• Rocznik: 2013
• Tom: 73
• Numer: 1
• Opis fizyczny: p.79-87,fig.,ref.

Twórcy

autor

Wagner D.-C.

• Fraunhofer Institute for Cell Therapy and Immunology, Leipzig, Germany
• Translation Center for Regenerative Medicine, Leipzig, Germany

autor

Scheibe J.

• Fraunhofer Institute for Cell Therapy and Immunology, Leipzig, Germany

autor

Glocke I.

• Fraunhofer Institute for Cell Therapy and Immunology, Leipzig, Germany

autor

Weise G.

• Fraunhofer Institute for Cell Therapy and Immunology, Leipzig, Germany
• Translation Center for Regenerative Medicine, Leipzig, Germany

autor

Deten A.

• Fraunhofer Institute for Cell Therapy and Immunology, Leipzig, Germany
• Translation Center for Regenerative Medicine, Leipzig, Germany

autor

Boltze J.

• Fraunhofer Institute for Cell Therapy and Immunology, Leipzig, Germany
• Translation Center for Regenerative Medicine, Leipzig, Germany
• Massachusetts General Hospital and Harvard Medical School, MA, USA

autor

Kranz A.

• Fraunhofer Institute for Cell Therapy and Immunology, Leipzig, Germany
• Translation Center for Regenerative Medicine, Leipzig, Germany

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