RESUMEN
The Bryan Alzheimer's Disease Research Center Rapid Autopsy Program at Duke University Medical Center obtains postmortem human brain tissue for experimental investigations. We evaluated 19 brains for RNA integrity and mRNA gene expression. Nine were from patients diagnosed with Alzheimer's disease, and ten were from nondemented controls. In all cases, the following variables were recorded: postmortem procurement delay (range, 1 hour and 10 minutes to 14 hours), pH of cerebrospinal fluid, premortem fever or sepsis, provision of supplemental oxygen in the agonal period, and temporal relation to time of death (either sudden death or protracted illness). Total RNA was extracted, quantified, and evaluated by agarose gel electrophoresis and quantitative gene expression analysis of 18S rRNA and edg-1 using TaqMan technology. All samples appeared to yield intact RNA without significant degradation, and expression of the edg-1 gene was detected by the real time reverse transcriptase polymerase chain reaction in all cases. We conclude that intact RNA can be obtained from postmortem human brain tissue, even in patients with severe premortem illnesses and delayed postmortem tissue procurement intervals. However, we caution that the successful expression of certain genes from postmortem brain tissue may require enhanced procurement efforts to maximize RNA integrity.
Asunto(s)
Encéfalo/metabolismo , ARN Mensajero/metabolismo , Receptores de Superficie Celular , Receptores Acoplados a Proteínas G , Anciano , Anciano de 80 o más Años , Femenino , Regulación de la Expresión Génica , Humanos , Proteínas Inmediatas-Precoces/genética , Masculino , Persona de Mediana Edad , Cambios Post Mortem , ARN Mensajero/genética , Receptores Lisofosfolípidos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Manejo de Especímenes/normas , Factores de TiempoRESUMEN
OBJECTIVE: To develop a system for retrieving brain tissue within 1 hour after death in an effective and useful manner. DESIGN: Nurse clinicians were employed as study co-ordinators and were available to families 24 hours each day. SETTING: Autopsies were performed at Duke University Medical Center, Durham, NC, from 1985 through 1995. PARTICIPANTS: Neuropathology faculty, fellows, and residents, autopsy technicians; and brain bank staff. RESULTS: Fifty-one rapid autopsies with a postmortem interval of less than 1 hour have been performed. Four of these were normal controls, three were disease controls, and 44 represented Alzheimer's disease patients. Tissue retrieved at rapid autopsy has been distributed to 93 research teams, 30 of these located at Duke University Medical Center. Many researchers have received multiple shipments of tissue. CONCLUSIONS: The Bryan Alzheimer's Disease Research Center Rapid Autopsy Program at Duke University Medical Center has been successful in retrieving tissue from individuals with dementia and also from controls within 1 hour of death. The critical features of the success of this program have been the use of nurse clinicians who work closely with patients and their families to ensure a successful autopsy at the time of death and the maintenance of a 24-hour call schedule for nurses and neuropathology staff. Similar programs can be implemented for experimental work into the pathogenesis of a wide variety of human diseases in which the examination of human tissue is required.
Asunto(s)
Autopsia , Encéfalo/patología , Obtención de Tejidos y Órganos/organización & administración , Anciano , Enfermedad de Alzheimer/patología , Autopsia/economía , Costos y Análisis de Costo , Demencia/patología , Familia , Humanos , Registros Médicos , Examen Neurológico , North Carolina , Cuidado Terminal , Factores de Tiempo , Bancos de Tejidos , Obtención de Tejidos y Órganos/economíaRESUMEN
The frequency of the allele for apolipoprotein E type 4 (epsilon 4) is increased in late-onset familial and sporadic Alzheimer's disease (AD). We have examined epsilon 4 frequencies in four distinct, normal, elderly control groups and, most importantly, in patients with amyloid-forming diseases whose epsilon 4 distributions were not previously known (Creutzfeldt-Jakob disease, familial amyloidotic polyneuropathy, Down's syndrome). There were no differences between any of these controls and published control series, cementing the relevance of epsilon 4 for late-onset AD. The increase in late-onset AD was confirmed in two new series.