Tubular aggregates (TAs) which have been recently observed in a few mouse myopathies are identical to those described in human diseases. In this study we show that TAs are also found in the skeletal muscle of almost all normal inbred mice strains. In these inbred strains of mice the presence of TAs is shown to be related to both age and sex. Nine different muscles were stained with the modified Gomori trichrome method to reveal the general morphology of the muscles. Anti-SERCA1 ATPase was used to confirm that the TAs were in fact accumulations of sarcoplasmic reticulum and anti-MyHC IIB to demonstrate that these accumulations were found exclusively in the type IIB muscle fibers. An ultrastructural study confirmed the observations revealed by light microscopy that the TAs were derived from the sarcoplasmic reticulum. TAs were never observed in female inbred mice and were only found in type IIB glycolytic muscle fibers of male inbred mice. Therefore when analyzing the effect of genetic knock out and knock in experiments on the muscle phenotype of transgenic mice one should be aware that the presence of these aggregates is a non-specific phenomenon induced by inbreeding.

The aim of the present study was to investigate whether ultrastructural features can be used as a guide to identify alpha- and gamma-motoneurons among the intermediate-size neurons of the peroneal motor nuclei. The peroneus brevis and peroneus tertius muscles of adult cats were injected with horseradish peroxidase, and motoneurons labeled by retrograde axonal transport were examined by electron microscopy. In both nuclei, the distributions of cell-body diameters, measured in the light microscope, were bimodal covering the range of 28-84 microns, with a trough around 50 microns. The sample of 25 motoneurons selected for the ultrastructural study included not only large (presumed alpha) and small (presumed gamma) neurons but also intermediate-size cell bodies with diameters in the 40-60 microns range. For each motoneuron, 2-5 profiles were reconstructed from ultrathin sections taken at 6-8 microns intervals. Synaptic boutons were counted and their lengths of apposition were measured. On the basis of three criteria, namely: (1) bouton types present on the membrane, (2) percentage of membrane length covered by synapses, and (3) the aspect of the nucleolus, all the examined motoneurons, including those with intermediate sizes, fell into one of two categories. Fourteen motoneurons, with cell-body diameters in a range of 55-84 microns, were contacted by all types of boutons (mainly S-type with spherical vesicles, F-type with flattened vesicles, and C-type with subsynaptic cistern); the synaptic covering of the somatic membrane was over 40% and the nucleus contained a vacuolated nucleolus. These were considered alpha-motoneurons. Eleven motoneurons, with only S and F boutons, a synaptic covering under 30%, a compact nucleolus and a cell-body diameter ranging between 28 and 50 microns, were considered gamma-motoneurons. No other combination of the three criteria was observed. These results show that unequivocal distinction of alpha- and gamma-motoneurons is possible in the peroneal nuclei, on the basis of morphological differences independent of cell-body size.

Poliovirus (PV) is the etiological agent of human paralytic poliomyelitis. Paralysis results from the destruction of motoneurons, a consequence of PV replication. However, the PV-induced process leading to the death of motoneurons is not well known. We investigated whether PV-induced central nervous system (CNS) injury is associated with apoptosis by using mice as animal models. Transgenic mice expressing the human PV receptor were infected intracerebrally with either the neurovirulent PV-1 Mahoney strain or a paralytogenic dose of the attenuated PV-1 Sabin strain. Nontransgenic mice were infected with a mouse-adapted PV-1 Mahoney mutant. DNA fragmentation was demonstrated in CNS tissue from paralyzed mice by visualization of DNA oligonucleosomal laddering and by enzyme-linked immunosorbent assay. Viral antigens and DNA fragmentation detected by the in situ terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end-labeling technique were colocalized in neurons of spinal cords from paralyzed mice. In addition, morphological changes characteristic of cells undergoing apoptosis were observed in motoneurons by electron microscopy. Thus, we show that PV multiplication and CNS injury during paralytic poliomyelitis are associated with apoptosis.

Poliovirus (PV) is the causal agent of paralytic poliomyelitis. Many survivors of the acute disease, after decades of clinical stability, develop new muscular symptoms called postpolio syndrome. It has been hypothesized that the persistence of PV in the spinal cord is involved in the etiology of this syndrome. To investigate the ability of PV to persist in the spinal cord after the onset of paralysis, we exploited a mouse model in which most animals inoculated with a mouse-adapted mutant survived after the onset of paralysis. Light microscopy and ultrastructural immunohistochemical studies and reverse transcription followed by nested PCR performed on spinal cord from paralyzed mice demonstrated that PV persisted in the mouse spinal cord for at least 12 months after the onset of paralysis. This mouse model provides a new tool for studying poliomyelitis evolution after the onset of paralysis.

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