carbocyanine-dye-diic12(3) and Disease-Models--Animal

Rett syndrome (RTT) is a neurodevelopmental disorder caused by mutations in the X-linked gene MECP2. Girls with RTT show dramatic changes in brain function, but relatively few studies have explored the structure of neural circuits. Examining two mouse models of RTT (Mecp2B and Mecp2J), we previously documented changes in brain anatomy. Herein, we use confocal microscopy to study the effects of MeCP2 deficiency on the morphology of dendrites and axons in the fascia dentata (FD), CA1 area of hippocampus, and motor cortex following Lucifer yellow microinjection or carbocyanine dye tracing. At 3 weeks of age, most (33 of 41) morphological parameters were significantly altered in Mecp2B mice; fewer (23 of 39) were abnormal in Mecp2J mice. There were striking changes in the density and size of the dendritic spines and density and orientation of axons. In Mecp2B mice, dendritic spine density was decreased in the FD (approximately 11%), CA1 (14-22%), and motor cortex (approximately 16%). A decreased spine head size (approximately 9%) and an increased spine neck length (approximately 12%) were found in Mecp2B FD. In addition, axons in the motor cortex were disorganized. In Mecp2J mice, spine density was significantly decreased in CA1 (14-26%). In both models, dendritic swelling and elongated spine necks were seen in all areas studied. Marked variation in the type and extent of changes was noted in dendrites of adjacent neurons. Electron microscopy confirmed abnormalities in dendrites and axons and showed abnormal mitochondria. Our findings document widespread abnormalities of dendrites and axons that recapitulate those seen in RTT.

Topics: Analysis of Variance; Animals; Axons; Carbocyanines; Dendritic Spines; Disease Models, Animal; Hippocampus; Isoquinolines; Male; Methyl-CpG-Binding Protein 2; Mice; Mice, Transgenic; Microinjections; Microscopy, Confocal; Microscopy, Electron; Motor Cortex; Neurons; Rett Syndrome

Removal of head and neck neoplasms, especially those of the parotid gland and those of the internal auditory canal and cerebellopontine angle, often requires microdissection of the facial nerve. Displacement or splaying of the nerve can make it difficult to identify facial nerve fibers and/or distinguish them from surrounding tissues. Here we tested a method of labeling the facial nerve with fluorescent lipophilic dyes as a method of providing intraoperative visual confirmation of nerve fibers.. The facial nerves of adult rats were retrogradely labeled with 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI), 3,3'-dioctadecyloxacarbocyanine perchlorate (DiO), or 3,3'-dilinoleyloxacarbocyanine perchlorate (Fast DiO) either by direct application to the nerve sheath or by microinjection into the facial muscles. The nerves were examined 30 days after dye application by means of a dissecting stereomicroscope equipped with epifluorescence filters.. Of the dyes tested, Fast DiO proved to be the most effective, providing labeling of the nerve sufficient to be seen with combined fluorescent and bright field stereomicroscopy. Nerve conduction studies indicated that fluorescent labeling did not adversely affect nerve function.. These results raise the possibility of using fluorescent lipophilic dyes to label nerves as a method of enhancing identification and distinguishing nerve fibers during surgery.

Topics: Animals; Carbocyanines; Disease Models, Animal; Electric Stimulation; Facial Nerve; Facial Nerve Injuries; Head and Neck Neoplasms; Humans; Monitoring, Intraoperative; Neural Conduction; Rats; Rats, Sprague-Dawley

Nerve injury represents a major cause of disability. In the peripheral nervous system, nerves have the capacity to regrow but within weeks after injury, it is impossible to clarify whether proper regeneration is under way or is failing. In this experimental study, we report on a novel tool to assess nerve outgrowth in vivo. After systemic application, the novel gadolinium-based magnetic resonance (MR) contrast agent Gadofluorine M (Gf) selectively accumulated and persisted in nerve fibers undergoing Wallerian degeneration causing bright contrast on T1-weighted MR images. Gf enhancement on MR imaging was present already at 48 hours within the entire nerve segments undergoing Wallerian degeneration, and subsequently disappeared from proximal to distal parts in parallel to regrowth of nerve fibers. Most importantly, Gf enhancement persisted in nonregenerating, permanently transected nerves. Our novel Gf-based MR imaging methodology holds promise for clinical use to bridge the diagnostic gap between nerve injury and completed nerve regeneration, and to determine the necessity for neurolysis and engraftment if spontaneous regeneration is not successful.

Topics: Animals; Carbocyanines; Disease Models, Animal; Ectodysplasins; Functional Laterality; Immunohistochemistry; Magnetic Resonance Imaging; Male; Membrane Proteins; Nerve Crush; Nerve Fibers; Nerve Regeneration; Organometallic Compounds; Rats; Rats, Wistar; Sciatic Neuropathy; Staining and Labeling; Time Factors; Tolonium Chloride; Wallerian Degeneration