Vitamin A and Retinoic Acid in Cognition and Cognitive Disease.

The history of vitamin A goes back over one hundred years, but our realization of its importance for the brain and cognition is much more recent. The brain is more efficient than other target tissues at converting vitamin A to retinoic acid (RA), which activates retinoic acid receptors (RARs). RARs regulate transcription, but their function in the cytoplasm to control nongenomic actions is also crucial. Controlled synthesis of RA is essential for regulating synaptic plasticity in regions of the brain involved in learning and memory, such as the hippocampus. Vitamin A deficiency results in a deterioration of these functions, and failure of RA signaling is perhaps associated with normal cognitive decline with age as well as with Alzheimer's disease. Further, several psychiatric and developmental disorders that disrupt cognition are also linked with vitamin A and point to their possible treatment with vitamin A or RA.

Use of fixatives for immunohistochemistry and their application for detection of retinoic acid synthesizing enzymes in the central nervous system.

Retinoic acid (RA) is a lipid signaling molecule that has a crucial role in growth and survival of neurons as well as regulation of neuronal plasticity in the central nervous system. Complete understanding of the distribution of RA is necessary to identify foci of RA signaling. However, RA itself is very difficult to detect by immunohistochemistry as there are few effective antibodies to this lipid and it can be difficult to fix in place in tissue. A set of retinaldehyde dehydrogenases (RALDHs) catalyze the last step of RA synthesis and their distribution can be used as a proxy for RA distribution. This protocol uses the example of RALDH2 expression in the motor neurons of the spinal cord as a demonstration of the approach and describes methods that can improve its effectiveness. Immunodetection is impacted by protein cross linking and protein denaturation as well as antigen/epitope masking by various fixatives. Finding a suitable fixative that preserves morphology and tissue structure by linking cellular component such as proteins and lipids in an indissoluble macromolecular network while keeping functional groups, including antigens, intact is essential. Here, we discuss fixatives in general and also describe a fixation protocol that allows detection of multiple antigens in the same section with a periodate-lysine-paraformaldehyde (PLP) fixative. This keeps tissue structure and antigen well preserved in the adult spinal cord to show RALDH2 expression in motor neurons.

Tissue localization of retinoic acid receptor (RAR) active drugs.

The retinoic acid (RA) signaling pathway is crucial for the control of embryonic development and also regulates function of several organ systems in the adult, including the central nervous system. The retinoic acid receptors (RARs) that mediate the majority of the functions of RA can promote proliferation, differentiation, morphogenesis and cell survival. Dysregulation of this signaling pathway has been considered in the pathophysiology of various diseases including neurodegenerative disorders such Alzheimer's disease and amyotrophic lateral sclerosis. Thus, drugs targeted to the RARs have been proposed as treatments for such diseases. Understanding how these drugs distribute in the body is essential to determine their potential effectiveness. However measuring tissue levels of what are often lipophilic drugs can be difficult. Here we describe an indirect measurement of RAR ligand tissue distribution after intraperitoneal injection into rodents that uses a sensitive RA reporter cell line.