a-stainless-protocol-for-high-quality-rna-isolation-from-laser

A Stainless Protocol for High Quality RNA Isolation from Laser Capture Microdissected Purkinje Cells in the Human Post-Mortem Cerebellum

Laser capture microdissection (LCM) is an advantageous tool that allows for the collection of cytologically and/or phenotypically relevant cells or ...

Columbia University

Columbia College

A review of the brain banking literature reveals a primary focus either on the factors that influence the decision to become a future donor or on the brain tissue processing that takes place after the individual has died (i.e., the front-end or back-end processes). What has not been sufficiently detailed, however, is the complex and involved process that takes place after this decision to become a future donor is made yet before post-mortem processing occurs (i.e., the large middle-ground). This generally represents a period of many years during which the brain bank is actively engaged with donors to ensure that valuable clinical information is prospectively collected and that their donation is eventually completed. For the past 15 years, the Essential Tremor Centralized Brain Repository has been actively involved in brain banking, and our experience has provided us valuable insights that may be useful for researchers interested in establishing their own brain banking efforts. In this piece, we fill a gap in the literature by detailing the processes of enrolling participants, creating individualized brain donation plans, collecting clinical information and regularly following-up with donors to update that information, and efficiently coordinating the brain harvest when death finally arrives.

Banking brains: a pre-mortem "how to" guide to successful donation.

Essential tremor (ET) patients develop more cognitive impairment and dementia than controls, although there are surprisingly few data on the neuropathological basis for cognitive changes in ET. In this postmortem study, we assessed tau and other pathologies in 26 ET cases and 73 controls (non-ET) (1:3 matching). The mean age = 88.6 years; 55% were cognitively normal, 24% had mild cognitive impairment (MCI), and 20% had dementia. We found similar burdens of pathology using Braak, β-amyloid and Lewy body assessments in ET and controls. In contrast, among cognitively normal subjects, ET cases had a higher number of NFT-positive neurons in the neocortex than controls (p < 0.001); the number of NFT-positive neurons in the medial temporal lobe was similar in these 2 groups (p = 0.22). Among subjects with MCI, ET cases also had higher numbers of NFT-positive neurons in the neocortex than controls (p < 0.001) but again, not in the medial temporal lobe (p = 0.55). Among subjects with dementia, the number of NFT-positive neurons was similar in ET cases and controls. Cognitive function correlated with quantitative neurofibrillary tangle counts in ET cases and controls. In the context of ET, pre-dementia tau burden is higher than in the absence of ET, suggesting a predisposition to tau pathology.

Quantitative Assessment of Pathological Tau Burden in Essential Tremor: A Postmortem Study.

The underlying biology of essential tremor (ET) is poorly understood. Purkinje cell (PC) loss has been observed in some studies, although this finding remains somewhat controversial. Basket cells are interneurons whose axonal collaterals form a plexus around PC soma. When there is PC loss, this basket plexus appears empty. We used dual immunohistochemical staining for calbindin D28k and glutamic acid decarboxylase to quantify "empty baskets" as an indirect and alternative method of detecting PC loss. Microscopic analyses on 127 brains included ET and a spectrum of motor neurodegenerative diseases (50 ET, 27 spinocerebellar ataxias [SCAs], 25 Parkinson disease, 25 controls). The median percentage of empty baskets in ET patients was 1.5 times higher than controls (48.8% vs 33.5%, p < 0.001) but lower in ET than in SCA1 (59.7%, p = 0.011), SCA2 (77.5%, p = 0.003), and SCA6 (87.0%, p < 0.001). PC loss is not a feature of SCA3, and the median percentage of empty baskets (30.1%) was similar to controls (p = 0.303). These data provide support for PC loss in ET and are consistent with the notion that ET could represent a mild form of cerebellar degeneration with an intermediate degree of PC loss.

Article	Year
Banking brains: a pre-mortem "how to" guide to successful donation. Cell and tissue banking\| PubMed ID: 30220002	2018
Quantitative Assessment of Pathological Tau Burden in Essential Tremor: A Postmortem Study. Journal of neuropathology and experimental neurology\| PubMed ID: 30476290	2019
A Quantitative Study of Empty Baskets in Essential Tremor and Other Motor Neurodegenerative Diseases. Journal of neuropathology and experimental neurology\| PubMed ID: 30590599	2019

Phyllis L. Faust

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