Table 2. Summary of IHC parameters of the herein utilized antibodies, and their roles in the body depending on subcellular location.
| Antibody | Dilution | + CTRL | CNS cell type | Significance of cellular pattern |
|---|---|---|---|---|
| polyclonal rabbit anti-Apaf-1 | 1:500 | Lung with lymphoma, rat | • Neurons | Cytoplasmic—up-regulated in the process of intrinsic apoptosis (including that in the developing brain); |
| • Glia (incl. pericytes) | Nuclear—associated to acute hypoxic conditions; | |||
| • Endothelia | Acellular perivascular leakage—mild hypoxia (likely a common phenomenon in diving mammals) may result in transient, physiological reduction of BBB integrity | |||
| polyclonal rabbit anti-DGK-ζ | 1:100 | Cerebellum, rat | • Neurons | Nuclear—common in morphologically healthy neurons; |
| • Glia | Nucleolar—potential interaction with Mdm2 and p53 proteins, which are implicated in nucleolar stress; | |||
| Cytoplasmic—association with pre-apoptotic state following ischemic-hypoxic episodes and auditory insults, may be neuroprotective and can also be associated with neuroplasticity | ||||
| polyclonal rabbit anti-Bcl-2 | 1:150 | Lymphonode, Canine | • Neurons | Cytoplasmic—low amounts expected in neonatal neurons and as an anti-apoptotic response to cell injury |
| • Glia | ||||
| monoclonal recombinant rabbit anti-Aβ | 1:1000 | Neocortex, canine neonate | • Neurons | Cytoplasmic—neurotoxic effect through disruption of calcium homeostasis, organelle and synaptic function; |
| • Glia | Nuclear—presumed regulation of apoptosis and potential indicator of neuroprotection against neurodegeneration; | |||
| • Endothelia | Extracellular plaques—presence positively correlated with cognitive decline in Alzheimer’s disease | |||
| Monoclonal mouse anti-NF200 | 1:400 | Cerebellum, rat | • Neurons | Cytoskeletal—reduced in cases of traumatic brain injury and hypoxia |
+CTRL: Positive control tissue. BBB: Blood-brain barrier.