Abstract
Background
In studies on cardiac arrest (CA)/resuscitation (R) injury, Purkinje cell degeneration was described, however, with inconsistent data concerning severity and time point of manifestation. Moreover, CA/R studies paid only limited attention to inhibitory stellate interneurons. To this aim, the hypothesis that cerebellar could be relatively resilient toward CA/R because of diverse cellular defense mechanisms including interaction with stellate cells was tested.
Methods
We examined rats with survival times of 6, 24, and 48 h, and 7 and 21 days in comparison with sham- and nonoperated animals. Thereby, we focused on the immunohistochemical expression of cfos, MnSOD, Bcl2, caspase 3, parvalbumin, calbindin D28 k, MAP2, IBA1, and GFAP, especially in the particular sensitivity to CA/R cerebellar lobule IX. Hippocampal CA1 degeneration was demonstrated by expression patterns of MAP2 and NeuN in combination with IBA1 and GFAP.
Results/Conclusions
Comparative analysis of hippocampal CA1 pyramidal cells and cerebellar Purkinje cells confirmed a relative resil-ience of Purkinje cells to CA/R. We found only a notable degeneration of Purkinje cell neuronal fiber network, which, however, not necessarily led to neuronal cell death. To induce significant Purkinje cell loss, a stronger ischemic trigger seems to be needed. As possible Purkinje cell-protecting mechanisms, we would propose: (1) activation of inhibitory stellate cells, shown by cfos, MnSOD, and Bcl2 expression, balancing out ischemia-induced excitation and inhibition of Purkinje cells; (2) translocation of the calcium-buffering system, shown by parvalbumin and calbindin D28 k expression, protecting Purkinje cells from detrimental calcium overload; (3) activation of the neuron–astrocyte cross talk, protecting Purkinje cells from over-excitation by removing potassium and neurotransmitters from the extracellular space; (4) activation of the effective and long-lasting MnSOD defense system; and (5) of the anti-apoptotic protein Bcl2 in Purkinje cells itself. Moreover, the results emphasize the limited comparability of animal CA/R studies because of the heterogeneity of the used experimental regimes.
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Abbreviations
- Bcl2:
-
B cell lymphoma 2
- CA/R:
-
Cardiac arrest/resuscitation
- DAPI:
-
6-Diamidino-2-phenylindole
- GCL:
-
Granule cell layer
- GFAP:
-
Glial fibrillary acidic protein
- HNF4A:
-
Hepatic nuclear receptor 4A
- IBA1:
-
Ionized calcium-binding adaptor molecule 1
- ID:
-
Integrated density
- IPPV:
-
Intermittent positive pressure ventilation
- LD:
-
Linear density
- MAP2:
-
Microtubule-associated protein 2
- ML:
-
Molecular layer
- MnSOD:
-
Manganese-dependent superoxide dismutase (mitochondrial SOD 2)
- NeuN:
-
Neuronal nuclei antibody
- PBS:
-
Phosphate-buffered saline
- PC:
-
Purkinje cell
- PCL:
-
Purkinje cell layer
- PFA:
-
Phosphate-buffered paraformaldehyde
- ROSC:
-
Return of spontaneous circulation
- ST:
-
Survival time
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Acknowledgements
The authors wish to thank Leona Bück for the excellent technical assistance.
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All authors have made substantive contributions to the study. GK conceived the study, was responsible for data analyses, writing the manuscript and preparing the illustration of results. TMNT carried out all immunohistochemical stainings. TE carried out surgery including post resuscitation care and tissue sampling. UE established the experimental model and supervised the animal experiments and critically reviewed the manuscript.
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This study was granted according to the requirements of the German Animal Welfare Act on the Use of Experimental Animals and the Animal Care and Use Committees of Saxony-Anhalt (permit number 42502-2-2-947 Uni MD).
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Keilhoff, G., Nguyen Thi, T.M., Esser, T. et al. Relative Resilience of Cerebellar Purkinje Cells in a Cardiac Arrest/Resuscitation Rat Model. Neurocrit Care 32, 775–789 (2020). https://doi.org/10.1007/s12028-019-00799-0
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DOI: https://doi.org/10.1007/s12028-019-00799-0