3-3-1- Cell and Tissue Response to Injury in the CNS 1. When provided with a histologic section, identify neurons, astrocytes, oligodendroglial cells, microglial cells and ependymal cells. Neurons are located in gray matter regions (unmeylinated cell bodies) and have that typical morphology of alarge nucleus, prominent nucleous, Nissl granules, and neuritis. In contrast, oligodendroglial cells are present in white matter regions and have small, round nuclei with no apparent cytoplasm Special stains can be used to definitively ID neurons of cells showing neuronal differentiation Bielchowsky silver stain identifies neurofilaments and inclusions, while Golgi silver stain identifies dendritic arborization. Immunoperoxidase stain IDs neurofilaments or synaptophysin (neuronal-specific synaptic protein). Luxol fast blue stains white matter blue. Immunhistochem (HC) Abs go against neurofilametnts and synaptophysin. H&E is good for staining meninges 2. Define four ways that neurons respond to injury (ischemic cell change, central chromatolysis, Wallerian degeneration, distal axonopathy). Ischemic cell change in neurons occurs in the form of red neurons, which is an active process that occurs 8-24 hours after the insult (thus, a person who drowns will not immediately have red neurons). The neuron shrinks, the cytoplasm becomes eosinophilic, and the nucleus shrinks to become darkly stained and is eventually lost. Intracellular changes include depletion of ATP, intracellular acidosis, impaired reuptake of glutamate by glial cells with resultant excitatory neurotransmitter damage in the nuron, accululation of intracellular calcium, and generation of free radicals. The process of cell death is irreversible Central chromatolysis occurs in neuronal cell bodies after sever injury to the axon (like a transection), most characteristically seen in large neurons, it presents as a swelling of the cell body, dissolution of Nissl granules, and migration ofthe nucleus ot the periphery. Unlike red neurons, central chromatolysis IS reversible and will revert to normalcy within a few months. ‘Wallerian degerneration occurs when the axon is transected. Distal to the transaction site, the axon and myelin sheath degenerated. The degenerating axon has impaired axonal transport, disappearance of neurofibrils, and breaking up of the axon into short fragments that will be phagocytosed. The process occurs much faster in the PNS (weeks) than in the CNS (months), and sprouting may occur in the PNS to regain function, while there is no regeneration in the CNS. Distal axonopathy occurs with neuronal toxicity or metabolic disorders in neuronal cells Ge. chemotherapy, alcoholism, diabetes, malnutrition). The most distal portion of the axon degenerates first, advancing towards the cell body. Regeneration may be possible with removal of the cause. Sensory, motor, DTRs, and autonomics are lost. Distal axonopathy often presents bilaterally and symmetrically 3. Describe the appearance and indicate a disease associated with each of five neuronalinclusions. ‘Types of inclusion bodies ‘© Neurofibrillary tangle. cytoplasmic argyrophilic inclusion found in Alcheimer's © Lewy body. cytoplasmic eosinphilic with “halo” inclusion found in Parkinson's. © Pick body: cytoplasmic argyrophilic inclusion found in Pick Disease (frontotemporal neuronal degeneration caused by dysfunctional tau protein accumulation) ‘© Negri body. cytoplasmic eosinophilic inclusion found in Rabies © Cowdry Type A nuclear eosinophilic inclusion found in herpes and CMV. 4. List three mechanisms by which myelin is reduced in amount in the CNS. IN which demyelinating disease are inclusions found in the oligodendroglia*? Myelin is formed in the CNS by oligodendroglial cells in the white matter (where each oligodendroglial cell can myelinate several axons), while myelin in the PNS is formed by Schwann cells (where each Schwann cell myelinates one axon) Myelin is visualized by Luxol Fast Blue (LFB) stain. It can result from 1. multiple sclerosis (MS): normal myelin is damaged by immunological mechanisms, which convert myelin into component lipids. Axons themselves are not damaged. 2. progressive multifocal leukoencephalopathy (PML): small plaques of demyelination develop due to death of oligodendroghtal cells. a. *The death of oligodendroglial cells is caused by papova virus infection of oligodendroglial cells, which results in homogenous, glassy, nuclear inclusions. 3. Leukodystrophies: genetic abnormalities resulting in the production of unstable myelin that breaks down, 5. Define astrogliosis (gliosis) and describe its appearance on histologic section. Astrocytes are present in both gray and white matter and can be IDed by GFAP (glial fibrillary acidic protein) staining via immunoperoxidase, They function in being a part of the BEB, having metabolic activity, and regulating the ionic environment of the brain. Astrogliosis refers to the reactive response of astrocytes to brain injury. It includes both proliferation and hypertrophy. Hypertrophied astrocytes, known as gemistocytes, have plump eocinophilic cytoplasm with accumulation of GFAP. Astrogliosis does not result in fibrosis Side note: astrocytes may develop inclusion bodies (called Rosenthal fibers) with infection of CMV. Rosenthal fibers appear eosinophilic and beaded, and are made of crystalline. This is similar to oligodendroglial cells being infected with papova virus, resulting in PML 6. Indicate the derivation of microglial cells in the CNS. All glial cells EXCEPT FOR MICROGLIAL CELLS are derived from the neuroectoderm. Microgliel cells are mesodermally derived from the bone marrow, then infiltrate into the developing brain along blood vessels.Once in the brain, they have slow tumover during life, appearing as small, elongated, dark- staining nuclei. 7. Define four ways in which microglial cells respond to injury (reactive microglial cell, macrophage response, microglial nodule, multinucleated giant cell). Microglial responses to injury: +, reactive microglial cell: activated microglial cells in response to brain injury or local immune response. Reactive cells are rod shaped with upregulated expression of MHC. molecules and inflammatory cytokines 2, macrophage response microglial cells différentiate into macrophages wien there is brain necrosis. Macrophages help phagocytose tissue debris, when they become lipid filled, they are called gitter cells Macrophages that hang around during brain injury come from monocytes. 3, microglial nodule these are formed by microglia encircling a single damaged neuron in encephalitis in order to phagocytose it. The process is known as neurophagia and results in the formation of a microglial nodule. 4. multinucleaded giant cell: occurs with HIV encephalitis, when groups of microglial cells accumulate in white matter and fuse to form the multinucleated giant cells. Changes are especially seen in AIDS dementia 8. Describe the typical response of ependymal cells to disruption. Ependymal cells line the ventricular system as a monolayer. They are joined by tight junctions and are ciliated ‘When disrupted, ependymal cells form rosettes that result from the proliferation of subependymal astrocytes adjacent to a site of ependymal cell loss. They begin as small protuberances known as ependymal granulations, which may progress to rosettes. 9. List three ways in which vasogenic edema differs from cytotoxic edema. Two types of cerebral edema ‘© Vasogenic edema: caused by loss of integrity of the BBB, which results in the etrance of excess water and accompanying solutes into the extracellular space. Expansion of the extracellular space, especially white matter, results in increased intracranial pressure © Cytotoxic edema: caused by toxic or metabolic events that disturb neuronal and glial cell membranes, resulting in intracellular accumulation of fuid in gray matter. There is usually NO MASS EFFECT.Learning Objectives: Cell and Tissue Response to Injury in the CNS 1. When provided with a histologic section, identify neurons, astrocytes, aligadendroglial cells, microglial cells and ependymal cells. « Neurons—large nucleus, prominent nucleolus, Niss! granules (RER), processes, arranged in layers. Pyramidal shaped cortical neurons. Myelin stains with Luxol Fast Blue. Immunohistochemistry can label neurons for neurofilament and synaptophysin. Golgi shows dendritic arborization. * Oligodendrocytes—small round nuclei with peri-nuclear clearing. Little cytoplasm on H&E. * Astrocytes—oval nuclei, many processes visible with GFAP stain (glial fibrillary acid protein) ‘* Microglia—small, elongated, dark staining nuclei. May differentiate into macrophages, eat lipids, and become glitter cells. May form nodules from neurophagia. May form multinucleated giant cells in HIV/AIDS. Ependymal Cells—monolayer lining ventricles. Tight junctions. Ciliated. Form rosettes after injury. ntral 2. _ Define four ways that neurons respond to injury (ischemic cell change, chromatalysis, Wallerian degeneration, distal axonopathy) * Ischemic Cell Change causes “Red Neurons” due to anoxia and loss of oxidative metabolism. 8-24 hours after injury, neuron shrinks, cytosol becomes eosinophilic, nucleus shrinks then darkens then disappears. Irreversible after several minutes of anoxia. Loss of ATP, acidosis, low glutamate uptake by glia with excitatory damage to neuron, intracellular calcium rises, free radicals rise. * Wallerian Degeneration is loss of distal axon due to axonal transection injury; degeneration of myelin sheath and axon distal to cut; atrophied muscle; proliferation of schwann cells eventually and regeneration of axon to renervate muscle. Faster in PNS with regeneration; slower in CNS without regeneration of function. * Central Chromatolysis is a central response to axonal damage, in large motor neurons. Swelling of cell body, dissolution of Niss! granules, migration of nucleus to periphery. Reversible; may be normal after a few months. « Distal Axonopathy is death of axon and myelin starting in most distal axon, but progresses toward cell body. Usually due to toxic chemicals like acrylamide, pesticide, solvents or from metabolic disease like diabetes, renal failure, alcoholism. Cell body cannot maintain metabolic needs of the axon. Occurs over time. 3. Describe the appearance and indicate a disease associated with each of five neuronal inclusions. 1, Neurofibrillary tangles—cytoplasmic, argyrophilic, Alzheimer’s disease. Bielschowsky silver stain, yellow with blackbrown