X-Message-Number: 33316 Date: Sun, 6 Feb 2011 13:42:44 -0800 (PST) From: Subject: an odd idea - fixation for cryonics? We assume that brain cells must be alive in order to store memories. Formaldehyde fixation is indeed highly destructive of the molecular basis of memory. However ethanol fixation apparently is not. Ethanol fixation may be compatible with cryonics, particularly since ethanol is also an effective cryoprotectant. Ethanol fixation would only be the treatment of last resort for political jurisdictions that ban or are otherwise hostile to cryonics. It would be superior to a straight freeze with no perfusion at all, since a straight freeze destroys the viability AND the morphology of brain cells, resulting in complete obliteration of the physical basis of memory. Brain Pathol. 2004 Apr;14(2):175-82. Comparison of ethanol versus formalin fixation on preservation of histology and RNA in laser capture microdissected brain tissues. Su JM, Perlaky L, Li XN, Leung HC, Antalffy B, Armstrong D, Lau CC. Laboratory of Molecular Neuro-oncology, Cancer Genomics Program, Texas Children's Cancer Center, Houston, Tex., USA. Abstract Although RNA can be retrieved from formalin-fixed, paraffin-embedded (FFPE) tissues, the yield is low, and the RNA is fragmented. Recent advances in gene expression profiling underscore the importance of identifying a fixative that preserves histology and mRNA. We demonstrated that, for immersion fixation of brains, 70% ethanol is superior to formalin for mRNA preservation. RNA yield from ethanol-fixed tissues was 70% of the yield from fresh frozen specimens, but only a negligible quantity was recovered from formalin-fixed tissues. RNA from ethanol-fixed brains showed integrity comparable to RNA from fresh frozen tissues, and RT-PCR using RNA from ethanol-fixed tissues was consistently successful. RNA from FFPE tissues composed of low-molecular weight fragments, and their use in RT-PCR failed repeatedly. The yield and quality of RNA from ethanol-fixed brains were unaffected after immersion at 4 degrees C for 2 weeks. In a blinded comparison to FFPE tissues, ethanol-fixed specimens were judged to show comparable histology and superior immunostaining. After laser capture microdissection (LCM), we failed to recover mRNA from FFPE tissues but retrieved mRNA from ethanol-fixed tissues for RT-PCR and cDNA microarray analysis. We conclude that 70% ethanol preserves RNA integrity and is suitable for expression profiling of brain tissues by LCM and cDNA microarray. PMID: 15193030 Hippocampus. 2010 May;20(5):584-95. >From learning to forgetting: behavioral, circuitry, and molecular properties define the different functional states of the recognition memory trace. Romero-Granados R, Fontan-Lozano A, Delgado-Garcia JM, Carrion AM. Division de Neurociencias, Universidad Pablo de Olavide de Sevilla, Carretera de Utrera, Km. 1, 41013-Sevilla, Spain. Abstract Neuropsychological analyses of amnesic patients, as well as lesion experiments, indicate that the temporal lobe is essential for the encoding, storage, and expression of object recognition memory (ORM). However, temporal lobe structures directly involved in the consolidation and reconsolidation of these memories are not yet well-defined. We report here that systemic administration of a protein synthesis inhibitor before or up to 4 h after training or reactivation sessions impairs consolidation and reconsolidation of ORM, without affecting short-term memory. We have also observed that ORM reconsolidation is sensitive to protein synthesis inhibition, independently of the ORM trace age. Using bdnf and egr-1 gene expression analysis, we defined temporal lobe areas related to consolidation and reconsolidation of ORM. Training and reactivation 21 days after ORM acquisition sessions provoked changes in bdnf mRNA in somatosensory, perirhinal, and hippocampal cortices. Reactivation 2 days after the training session elicited changes in bdnf and egr-1 mRNA in entorhinal and prefrontal cortices, while reactivation 9 days post-training provoked an increase in egr-1 transcription in somatosensory and entorhinal cortices. The differences in activated circuits and in the capacity to recall the memory trace after 9 or 21 days post-training suggest that memory trace suffers functional changes in this period of time. All these results indicate that the functional state of the recognition memory trace, from acquisition to forgetting, can be specifically defined by behavioral, circuitry, and molecular properties. 2009 Wiley-Liss, Inc. PMID: 19603520 Rate This Message: http://www.cryonet.org/cgi-bin/rate.cgi?msg=33316