In situ demonstration of mature oligodendrocytes and their processes: An immunocytochemical study with a new monoclonal antibody, Rip

Abstract

This paper introduces "Rip" a monoclonal antibody that produces relatively complete staining of oligodendrocytes and their processes in the adult central nervous system (CNS). The distribution of Rip immunoreactivity coincides with that of myelinated axons in both the spinal cord and the cerebellum. In addition, double-immunolabeling experiments demonstrate that Rip stains processes containing myelin basic protein but does not stain processes that express glial fibrillary acidic protein. These results indicate that Rip selectively stains oligodendrocytes but not astrocytes. Moreover, individual Rip-stained oligodendrologial somata and their cytoplasmic processes were observable at both the light microscopic and electron microscopic level when the staining of myelin was reduced. This was accomplished by omitting detergents from antibody incubation steps. Rip-stained oligodendrocytes have multiple processes of varying thickness, some of which end in close proximity to myelin sheaths. These immunostained profiles, reminiscent of those observed in oligodendrocytes stained by Golgi methods, are unique to Rip and indicate that its immunoreactivity is distinct from that of existing serological markers for oligodendrocytes.

Selective temporal and regional alterations of Nogo-A and small proline-rich repeat protein 1A (SPRR1A) but not Nogo-66 receptor (NgR) occur following traumatic brain injury in the rat

“…Nogo-A is present in all stages of oligodendrocyte differentiation (Huber et al, 2002;Wang et al, 2002b) and is believed to mediate inhibition of neurite outgrowth and axonal regeneration through binding to and activation of neuronal NgR in juxtaposed axons (Wang et al, 2002b;. In the present report, RIP was used to label oligodendrocytes as previously described (Friedman et al, 1989;Nguyen and Pender, 1999;Wennstrom et al, 2004), and we observed an increased number of Nogo-A (+)/RIP (+)cells in ipsilateral white matter following FP brain injury. Although an increased number of glial cells have been observed in the ipsilateral white matter tracts following FP brain injury (Graham et al, 2000), our data are the first to suggest that the number of oligodendrocytes may increase in selected regions following TBI, as previously demonstrated in models of cerebral ischemia (Dewar et al, 2003;Zaidi et al, 2004), spinal cord injury (McTigue et al, 2001) and cortical resection brain injury (Ludwin, 1984).…”

α-Tanycytes of the adult hypothalamic third ventricle include distinct populations of FGF-responsive neural progenitors

“…5e,f). Both early-and late-passage neurospheres differentiated after growth factor reduction, as assessed by the presence of RIP þ ive oligodendrocytes 34 , GFAP þ cells with astrocytic morphology 35 , TUJ1 þ ve immature neurons 36,37 and growth hormone releasing hormone (GHRH) þ ive hypothalamic neurons 32 (Fig. 5g).…”

Stimulation of Adult Oligodendrogenesis by Myelin-Specific T Cells

“…Oligodendrocytes and myelin were visualized using monoclonal mouse anti-rat Rip antibody (Developmental Studies Hybridoma Bank, University of Iowa, Iowa City, IA) recognizing 2=, 3=-cyclic nucleotide 3=-phosphodiesterase. 32,33 The antibody was biotinylated by Karsten Skjødt, University of Southern Denmark (Odense) and used at a concentration of 0.05 mg/mL. NG2 and CNP stainings were counterstained using toluidine blue to facilitate cell counting.…”