Lipofuscin, the so-called ageing pigment, is formed by the oxidative degradation of cellular macromolecules by oxygen-derived free radicals and redox-active metal ions. Usually it accumulates in post-mitotic, long-lived cells such as neurons and cardiac muscle cells. In contrast, it is rarely seen in either normal or diseased skeletal muscle fibres. In this paper, we report that lipofuscin accumulates at an early age in both human and murine dystrophic muscles. Autofluorescent lipofuscin granules were localized, using confocal laser scanning microscopy and electron microscopy, in dystrophin-deficient skeletal muscles of X chromosome-linked young Duchenne muscular dystrophy (DMD) patients and of mdx mice at various ages after birth. Age-matched normal controls were studied similarly. Autofluorescent lipofuscin granules were observed in dystrophic biceps brachii muscles of 2-7-year-old DMD patients where degeneration and regeneration of myofibres are active, but they were rarely seen in age-matched normal controls. In normal mice, lipofuscin first appears in diaphragm muscles nearly 20 weeks after birth but in mdx muscles it occurs much earlier, 4 weeks after birth, when the primary degeneration of dystrophin-deficient myofibres is at a peak. Lipofuscin accumulation increases with age in both mdx and normal controls and is always higher in dystrophic muscles than in age-matched normal controls. At the electron microscopical level, it was confirmed that the localisation of autofluorescent granules observed by light microscopy in dystrophin-deficient skeletal muscles coincided with lipofuscin granules in myofibres and myosatellite cells, and in macrophages accumulating around myofibres and in interstitial connective tissue. Our results agree with previous biochemical and histochemical data implying increased oxidative damages in DMD and mdx muscles. They indicate that dystrophin-deficient myofibres are either more susceptible to oxidative stress, or are subjected to higher intra- or extracellular oxidative stress than normal controls, or both.

Three transgenic mouse lines, ICR-PVRTg1, ICR-PVRTg5, and ICR-PVRTg21, which are susceptible to poliovirus, have been established by introducing the human gene for poliovirus receptor (PVR) into the genome of mouse strain ICR. Genetic characterizations of the PVR gene were carried out on these mouse lines to define the approximate copy number, insertion site, and expression of the transgene in the central nervous system (CNS). The transgene was integrated in the chromosome 4, 12, and 13 of ICR-PVRTg1, ICR-PVRTg5 and ICR-PVRTg21 mice, respectively, and was stably transmitted to progeny mice. ICR-PVRTg1 appeared to have the most abundant copy numbers of the transgene and showed the highest level of PVR mRNA and membrane associated PVR protein in the CNS among the three mouse lines. Those in ICR-PVRTg21 and ICR-PVRTg5 were at intermediate and lowest levels, respectively. In the CNS, PVR mRNA was detected at high levels only in neurons of the spinal cord and brain stem where poliovirus can replicate, suggesting that the PVR mRNA expression confers cell specificity to poliovirus in the CNS. ICR-PVRTg1 and ICR-PVRTg5 showed the highest and the lowest sensitivity to poliovirus, respectively, whereas ICR-PVRTg21 was in-between. These results may suggest that poliovirus sensitivity of the mice is attributed to relative levels of PVR expression.

Abstract:We have immunocytochemically shown a significant reduction in the amount of laminin M (or merosin; a tissue-restricted basal lamina protein expressed in striated muscle, Schwann cells, and placental trophoblast) in the skeletal muscle of Fukuyama type congenital muscular dystrophy (FCMD).1) To inquire into the role of laminin M in the process of muscular dystrophies, we examined laminin M in several animal models that cause muscular dystrophy. Immunofluorescent, immunoblotting, and electron microscopic analyses have revealed that laminin M is missing from skeletal and cardiac muscles and peripheral nerve in the affected homozygous C57BL/6J-dy/dy mice, but not in the non-affected heterozygous Dy/dy and the other dystrophic animal models including mdx mice, BIO 14.6 hamsters, and line 413 chickens. In the dy/dy mice, laminin M mRNA is not detected by Northern blotting, but becomes detectable by RT-PCR amplification. Other components of the basal lamina such as laminin B, beta-integrin, type IV collagen, and fibronectin are normally expressed in all animals examined, including the dy/dy mice. These observations strongly suggest that laminin M defect is primarily responsible for the pathogenesis of muscle fiber damage and dysmyelination of the dystrophic dy/dy mice.

highlands at an altitude of 6,100 m and are typical animals adapted to high-altitude among mammals of the world [1]. These animals have a long history of habitation at high-altitudes and are considered to be "animals completely adapted to high-altitude" because of their physiological and morphological traits that are well adapted to high-altitude environments.When animals are exposed to a high-altitude environment over a long period, the pulmonary artery pressure increases because of an increase in the hematocrit (Ht) associated with an increase in the red blood cell count and hypoxic pulmonary vasoconstriction (HPV), causing pulmonary hypertension and right Japanese Journal of Physiology, 53, 377-384, 2003 Key words: blue-sheep, high-altitude, pulmonary hypertension, right ventricular hypertrophy. Abstract:The blue-sheep, pika, and yak live in the Tibetan highlands at an altitude of 6,100 m and are typical mammals adapted to high-altitudes. These animals have a long history of habitation at high-altitudes and are considered to be "animals completely adapted to high-altitudes" because of their physiological and morphological traits that are well adapted to high-altitude environments. To evaluate the physiological characteristics of high-altitude adaptation in the bluesheep, changes in the pulmonary hemodynamics during exposure to simulated-altitudes at 0, 2,300, and 4,500 m were examined by means of a climatic chamber in Qinghai Province, China (altitude 2,300 m). Seven blue-sheep inhabiting the mountains (3,000 m) of Qinghai Province, China, were compared with 5 pigs raised in the same area as controls. The primary items of measurement were the body weight (BW), systemic arterial pressure (Psa), pulmonary artery pressure (Ppa), hematocrit (Ht), left ventricular weight (LVW), right ventricular weight (RVW), and blood gas profile. The principal findings of this study are: (1) Ht, an index of right ventricular hypertrophy (RVW/LVW), and oxygen consumption (V O 2 ) were significantly lower in the blue sheep compared with the pigs; (2) When the animals were exposed to simulated-altitudes at 0, 2,300, and 4,500 m, Ppa increased significantly in tandem with altitude elevation in both species, but the increases were significantly smaller in the blue-sheep; and (3) Ppa/Psa, an index of the right ventricular load, increased with the altitude in both species, but the increases were smaller in the blue sheep. From these observations, low Ht and RVW/LVW and significant attenuation of hypoxic pulmonary vasoconstriction (HPV) in the blue-sheep is considered to be characteristics of animals completely adapted to high-altitudes, such as the pika.