A Light and Electron Microscope Study of Long-Term Organized Cultures of Rat Dorsal Root Ganglia

Abstract

Dorsal root ganglia from fetal rats were explanted on collagen-coated coverslips and carried in Maximow double-coverslip assemblies for periods up to 3 months. These cultured ganglia were studied in the living state, in stained whole mounts, and in sections after OsO4 fixation and Epon embedment. From the central cluster of nerve cell bodies, neurites emerge to form a rich network of fascicles which often reach the edge of the carrying coverslip. The neurons resemble their in vivo counterparts in nuclear and cytoplasmic content and organization; e.g., they appear as "light" or "dark" cells, depending on the amount of cytoplasmic neurofilaments. Satellite cells form a complete investment around the neuronal soma and are themselves everywhere covered by basement membrane. The neuron-satellite cell boundary is complicated by spinelike processes arising from the neuronal soma. Neuron size, myelinated fiber diameter, and internode length in the cultures do not reach the larger of the values known for ganglion and peripheral nerve in situ (30). Unmyelinated and myelinated nerve fibers and associated Schwann cells and endoneurial and perineurial components are organized into typical fascicles. The relationship of the Schwann cell and its single myelinated fiber or numerous unmyelinated fibers and the properties of myelin, such as lamellar spacing, mesaxons, Schmidt-Lanterman clefts, nodes of Ranvier, and protuberances, mimic the in vivo pattern. It is concluded that cultivation of fetal rat dorsal root ganglia by this technique fosters maturation and long-term maintenance of all the elements that comprise this cellular community in vivo (except vascular components) and, furthermore, allows these various components to relate faithfully to one another to produce an organotypic model of sensory ganglion tissue.

Satellite glial cells in sensory ganglia: from form to function

“…We used this method for patch clamp recordings from SGCs in mouse trigeminal ganglia [182]. Culturing intact ganglia (organ culture) is a useful model, as the relations among cells appears to be preserved over periods of months [22], and therefore this method might offer advantages for investigating SGCs. Isolated intact sensory ganglia should be the first choice for studying SGCneuron interactions as there is minimal tissue disruption.…”

Fine Structure of Neurons in the Spinal Ganglion of Slow Loris (<i>Nycticebus coucang coucang</i>)

“…Furthermore, the evidence with regards to the presence of such membrane-particle complexes in the nervous tissue is very limited. The only available reports are of BUNGE et al (1967) andPANNESE (1969) in which the authors have described the presence of such complexes in vitro and in vivo respectively.…”

“…Several studies have been made on the fine structure of spinal ganglia in various mammals (BEAMS et al, 1952;DAWSON, HOSSACK and WYBURN, 1955;PALAY and PALADE, 1955;DUNCAN and NAIL, 1960;HIRAOKA and BREEMEN, 1963;TENNYSON, 1964;BUNGE et al, 1967;PANNESE, 1969) but there is no reference to the ultrastructure of the spinal ganglion cells of the slow loris (Nycticebus coucang coucang), a prosimian primate.…”

Label-free Imaging of Schwann Cell Myelination by Third Harmonic Generation Microscopy

“…3D). The classic funnel shape, familiar from previous electron and light microscopy (13,14), had an inclination angle of ∼10° (Fig. 3E).…”