Neurofilament in feline primary afferent neurones: a quantitative immunocytochemical study

Two populations of cat L4 dorsal root ganglion (DRG) neurones were apparent from their Nissl staining with Toluidine blue. One had neurones of all sizes and the other had predominantly small neurones. The size distribution of neuronal profiles in the two populations overlapped and both were approximately normal. They corresponded to the light (L) and small dark (SD) cell populations previously described in rat DRGs. These neurones were examined with four antibodies to neurofilament: RT97, NFH, 155 and anti-68kD. RT97 is specific for the phosphorylated form of the 200 kDa subunit; NFH recognises both phosphorylated and non-phosphorylated forms of this subunit; 155 and anti-68 kDa recognise the 155 kDa and 68 kDa subunits respectively. The clearest differential labelling was seen with NFH and RT97 and this labelling was compared with cell size. High intensity NFH labelling was in population of neuronal profiles of all sizes and low intensity labelling in a population of predominantly small neuronal profiles. These populations corresponded respectively to the L and SD populations seen with toluidine blue staining. In the rat, these populations can be demonstrated by both NFH and RT97. In contrast in the cat, high intensity RT97 labelling was seen in only 75% of the L neuronal profiles defined with NFH and was also seen in some SD neuronal profiles defined with NFH. It is thus proposed that L and SD cell types are present in the cat DRG and can be demonstrated using the anti-neurofilament marker, NFH.     

Desmoplastic ganglioglioma: report of two non-infantile cases

Summary Two supratentorial desmoplastic gangliogliomas arising in a 15-year-old boy and a 25-year-old man are reported. Both tumors reached the brain surface and exhibited large cysts. They showed intense desmoplasia and tumor cells of astrocytic and ganglionic differentiation. In one case the ganglionic nature was only demonstrable by immunohistochemistry. Such neoplasms can no longer be regarded as exclusively infantile brain tumors.     

A subpopulation of chicken primary sensory neurons defined by complete co-localization of Peripherin- and ovalbumin-immunoreactivities

In a previous study, we have demonstrated that an ovalbumin-like antigen is present within approximately one-half of all neurons of chicken spinal ganglia. The current study demonstrates this antigen co-localizes absolutely with neural intermediate filament protein (Peripherin) in small to medium-sized neurons of spinal ganglia. While the function of ovalbumin in neurons is unknown, its precise co-localization with Peripherin suggests a functional role restricted to neurons of a defined phenotype.     

Neurofilament distribution and organization in the myelinated axons of the peripheral nervous system

The nature of neurofilament organization within the axonal cytoskeleton has been the subject of controversy for many years. Previous reports have suggested that neurofilaments are randomly distributed in the radial dimension of the myelinated axon. Randomness of distribution implies that there is no interaction between neurofilaments, while order in distribution suggest the presence of forces between neurofilaments. To address the issue of randomness vs. order, we evaluated neurofilament distribution by two different statistical approaches—nearest-neighbor distance and the Poisson tile-counting method. Neurofilament nearest-neighbor distances in a myelinated axon differ from nearest-neighbor distances of a set of random points with similar density (40.6 ± 7.0nm vs.30.7 ± 12.9nm, P < 0.0001). The Poisson tile-counting method also indicated that neurofilament distribution is different from a random distribution, under conditions of appropriate tile size and masking of other organelles. To further characterize the distribution of neurofilaments, we compared the relationship between nearest-neighbor distance and density for three sets of data: evenly spaced points, randomly distributed points and measured neurofilament coordinates. Neurofilaments do not conform to either evenly spaced or random distribution models. Instead, neurofilament distribution falls into an intermediate position between evenly spaced and random distributions. This study also demonstrates that the nearest-neighbor distance method of assessing neurofilament distribution offers several technical and theoretical advantages to the Poisson tile-counting method.     

Mature sensory neuron-derived hybrid cell line expressing NGF-dependent neurite extension

We established a hybrid cell line which extended neurites in the presence of nerve growth factor by fusion between adult mouse sensory neurons and neuroblastoma C1300 cells using emetine and actinomycin D. The serine-phosphorylated 200 and 160 kDa neurofilament proteins were detected in the hybrid cells. In comparison, C1300 cells expressed both subunit of non-phosphorylated neurofilaments at serine residues.     

Neuron-specific enolase and neurofilament protein as markers of differentiation in medulloblastoma

Immunocytochemical localization of neuron-specific enolase was performed attempting evaluation for neuronal cell differentiation in medulloblastoma. Twenty-seven cases of human medulloblastomas were stained with anti-neuron-specific enolase and antineurofilament protein serum using the peroxidase-antiperoxidase technique. All medulloblastomas showed neuron-specific enolase immunoreaction but only few had neurofilament protein-positive cells. These results suggest that a practically universal tendency towards neuronal cell differentiation occurs in medulloblastomas and that synthesis of neuron-specific enolase takes place before sufficient amounts of neurofilament protein are produced to become immunocytochemically detectable.     

Increased calpain content and progressive degradation of neurofilament protein in spinal cord injury

Spinal cord injury was induced in rat by weight drop. The extent of degradation of neurofilament proteins in the lesion following trauma was examined and served as a measure of calpain activity. Calpain was identified in the samples by myelin mealpain antibody and the content was estimated from the immunoblot. There was progressive degradation of both 68 kDa and 200 kDa neurofilament proteins in the cord lesion at intervals after injury. At 30 min after injury there was 20% degradation of both neurofilament proteins while the breakdown of 68 kDa and 200 kDa NFRs amounted to more than 60% at 24 h and beyond. Calpain content progressively increased in the lesion by 22% at 30 min to 91% at 4 h after trauma compared to control and then decreased but remained elevated for up to 72 h following injury. These results suggest that calpain is a primary responder synthesized early in injury and involved initially in the breakdown of cytoskeletal proteins in spinal cord trauma. Later in the injury cascade, increased calpain activity is derived from inflammatory as well as endogenous cells supporting a pivotal role for calpain throughout the process of secondary and evolving tissue damage in spinal cord trauma.     

Age-related alterations in immunoreactivity of the midsized neurofilament subunit in the brainstem reticular formation of the cat

In the present study, we compared the immunoreactivity of the midsized subunit of neurofilaments (NF-M) in the brainstem reticular formation of adult and old cats. There was a dramatic decrease in immunoreactivity in most reticular nuclei in the old cats. The most obvious reduction in these regions occurred in dendritic arborizations. In contrast, a small number of nuclei showed a slight increase in immunoreactivity in the aged animals. The age-related changes in immunoreactivity indicate that there is an alteration of NF-M content in reticular neurons and their processes in old age. Such changes in NF-M content may be the basis for the alterations in the morphology of reticular neurons in aged animals.     

A new case of Pick's disease

Summary The fine structure of a cortical frontal biopsy of Pick's disease is described. Pick bodies appear made of unbranched 120 Å neurofilaments, sometimes clustered in geometrical pattern. Post-mortem examination, performed 8 years later, reveals typical lesions. The characteristics of Pick bodies are discussed.Part of this study was presented at the VIIIth International Congress of Neuropathology (Washington, Sept. 1978)This work was supported by INSERM, grant no. 76.5-206-6     

The axon reaction in motor and sensory neurones of mice studied by a monoclonal antibody marker of neurofilament protein

Following unilateral sciatic nerve crush in mice, changes in the neurofilament content of neuronal perikarya were studied, using a monoclonal antibody to neurofilament protein (RT97). In the spinal cord, anterior horn motor neurones, normally unstained, showed a positive staining reaction with immunoperoxidase on the operated side. This reaction was short lived and maximal on the 11th post-operative day. In spinal ganglia, the proportion of positively staining sensory neurones showed an earlier but otherwise similar increase. In both cases, the response was well defined and contrasted with the changes on Nissl staining, which were markedly different in the two populations of neurones. In the nerve crush region, although regenerating axons were visible with silver staining only 5 days post-operatively, neurofilament protein was not demonstrated in these axons until several days later, after the peak perikaryal increase. These results suggest that an increase in perikaryal neurofilament protein is a consistent and quantifiable event following distal axon trauma, possibly indicating either synthesis of protein subunits or repolymerization of neurofilaments prior to their transport distally down the regenerating axons. The findings may be useful in identifying neurones with distal axon lesions in experimental and other neuropathological material.