Fig 1.
Illustration of the experimental setup.
The positions of the cranial window for laser speckle contrast imaging, the silver wire for EEG recording and the catheters for blood injection and ICP assessment are shown.
Fig 2.
A: Diagram of sample size distribution: 32 animals were randomly allocated to four different groups for blood sampling for inflammation markers (IL-6 and TNF-α) and corticosterone; after subtracting drop-outs from surgery and recording phase, a total of 21 animals were included in the final analysis. Subarachnoid hemorrhage (SAH) was induced by injection of 500μl arterial blood within 1 minute into the cisterna magna, sham animals did not receive any injection. B: Time points for blood sampling for inflammatory cytokine and corticosterone analysis: The diagram shows the time points (x) when blood was withdrawn within basic surgery (anaesthesia, tracheotomy, ventilation, A./V. femoralis cannulation), head fixation and cranial preparation (turning to prone position, fixation with ear bars in stereotactic frame, cranial window preparation, trepanations for EEG and ICP recording, cannula for icv blood injection) followed by the measurement period; red flash marks the time point of SAH induction.
Table 1.
Physiological parameters achieved from blood gas analysis, rectal temperature probe and pulse oximeter.
Fig 3.
Box plots of IL-6 and TNF-α in blood samples from SAH (black) and sham (grey) animals.
Data are presented as boxplot diagrams; the whiskers represent the minimum and maximum values, the dots show single points. Data were analyzed using repeated measures 2-way ANOVA followed by multiple comparisons tests: Sidak´s (comparisons between the groups; (A, B) IL-6 (A) and TNF-α (B) sham: n = 6; SAH: n = 5) or Tukey´s ((C, D) for IL-6, comparisons within the group; only significances in respect to baseline are depicted); * p < 0.05; ** p < 0.01; dotted line: minimum detectable dose (IL-6: 21pg/ml, TNF-α 5pg/ml).
Fig 4.
Box plots of corticosterone in blood samples from SAH (black) and sham (grey) Corticosterone was analysed at predefined sampling time points within the surgery and during the recording phase; Data are presented as boxplot diagrams; the whiskers represent the minimum and maximum values, the dots show single points. Data were analyzed using repeated measures 2-way ANOVA followed by Sidak´s multiple; SAH: n = 5, sham: n = 5; * p < 0.05; dotted line: minimum detectable dose 6.1ng/ml.
Fig 5.
Western blot analysis of TNF-α (A) and IL-6 (B) in 3 different brain regions Upper part: representative protein bands detecting TNF-α or IL-6. Lower part: box plots of quantitative analysis from SAH (black) and sham (grey); data were analysed using 2 way ANOVA followed by Tukey´s (within groups) or Sidak´s (between groups) multiple comparisons tests. brain tissue was harvested after euthanasia at the end of the recording phase at 6h. * p < 0.05; ** p < 0.01.