Makoto Sugimura

Extracellular matrix of the superior olivary nuclei in the dog

SummaryThe extracellular matrix around nerve cell bodies in canine lateral and medial superior olivary nuclei was examined by conventional electron microscopy, Golgi impregnation and histochemical techniques. Each neuron is surrounded by a region of myelin-free neuropil embedded amongst the myelinated fibres of the trapezoid body. In the myelin-free neuropil there are astrocytes, axons, synaptic boutons and extracellular matrix. The extracellular matrix fills the spaces between slender axons near the terminals, synaptic boutons and glial processes, but not the synaptic cleft. Golgi impregnation selectively stains the perineuronal nets which cover some or all of the nerve cell bodies and dendrites. The Golgi-EM method revealed that the impregnated profiles of the nets are restricted to the extracellular matrix. Synaptic boutons are situated in the holes of the perineuronal nets. Peanut (PNA) and soybean (SBA) agglutinins bound the extracellular matrix but not the synaptic boutons, glial processes, nerve cell bodies or basal lamina of blood capillaries. Light microscopic immunohistochemistry of the glial fibrillary acidic protein (GFAP) and S-100 protein did not stain a layer corresponding to the extracellular matrix and synapses but showed an intensely positive reaction immediately outside this layer. These data suggest the existence of a unique microenvironment associated with glycoconjugates around nerve cell bodies in canine superior olivary nuclei.

Isolation and antibody prevalence of a parapoxvirus in wild Japanese serows (Capricornis crispus).

An epizootic suspected to be caused by parapoxviruses occurred in winter, 1984–85, among wild Japanese serows (Capricornis crispus) with nodular or papular lesions in Gifu Prefecture, Japan. Virus isolations were attempted on 30 animals using bovine fetal testicle cell cultures. Viral agents growing with cytopathic effects were isolated from six animals. The agents were identified as parapoxvirus from electron microscopic findings and physico-chemical characteristics. Antibody prevalence to the isolated agent, named S-1, was examined by enzyme-linked immunosorbent assay. No antibodies were detected among 153 sera obtained during two winters 1981–82 and 1982–83, but one of 189 sera collected in the winter of 1983–84 was positive. In contrast, 75 (32%) of 237 sera had antibodies to S-1 in 1984–85, when the disease was first detected. Antibody prevalences of 1984–85 were highest (39%) in December 1984, the first month of animal capture, and declined gradually to 20% in March 1985.

Horn growth and reproductive history in female japanese serow

To elucidate the relationship between horn growth and reproduction, we measured the annual horn growth increments and histologically examined the ovaries of 132 female Japanese serow ( Capricornis crispus ). The number of small growth increments observed in each horn coincided with the number of elastoid bodies, thought to be the retrograde corpora lutea of pregnancy, in the ovaries of 77.6% of females ≥4.5 years old. Therefore, horn growth measurements can be used to trace the reproductive history of females. Horn growth measurements were used to estimate age-specific and annual fertility rates in a sample of 172 females from a wild population. Age-specific fertility rate peaked at 6 years of age (81.0%). Fertility rate of females ≥3 years old averaged 71.5% for 1981–83. Age when first reproduction occurred ranged from 2 to 5 years. Annual probability of reproducing was greatest for the female group that reproduced first at 3 years old.

Polyglucosan bodies in the digestive tract of the aged dog.

SummaryIn our recent studies on aging phenomena in animals, polyglucosan bodies (PGB) were found within the smooth muscle in the digestive tract of aged dogs without neurologic signs. PGB were basophilic, round, or oval bodies which appeared to have a homogenous or concentric shape. Their histochemical properties were characterized by the presence of glucose polymers (polyglucosan). Electron microscopy revealed that PGB were composed mainly of irregularly clustered, short branching filaments measuring about 90 Å in width. PGB were histochemically and ultrastructurally identical to the previously reported Lafora-like bodies in the CNS of aged dogs. PGB were found in all aged dogs and were disseminated throughout the digestive tract, especially in the cecum.

The Preputial Gland of the Japanese Serow Capricornis crispus: infrastructure and Lectin Histochemistry

Preputial glands of the Japanese serow were found to consist of the modified sebaceous gland and apocrine gland in both sexes. In the modified sebaceous gland, cells transformed gradually toward the lumen from plentiful cytoplasm to a sponge-like network. Lipid droplets were observed in mitochondria of differentiating cells. By lectin histochemistry, differentiating cells were stained with Arachis hypogaea (PNA), Triticum vulgaris (WGA) and Canavalia ensiformis (Con A) lectins, while secretory cells of the apocrine gland revealed various stainings with PNA, Glycine max, Dolichos biflorus, WGA and Con A lectins. These findings suggest that glycoconjugates may have an active function in the preputial gland.

Lectin histochemistry of canine polyglucosan bodies

SummaryLectin histochemistry was investigated to identify sugar residues of the polyglucosan bodies of canine brain, spinal cord and caecum. The polyglucosan bodies in the brain and spinal cord stained with concanavalin A (ConA) but not with soybean agglutinin, wheat germ agglutinin, peanut agglutinin,Dolichos biflorus agglutinin,Ricinus communis agglutinin andUlex europaeus agglutinin. Caecum, polyglucosan bodies, however, did not stain with any of the seven lectins employed. After periodate oxidation, paradoxical ConA staining was observed in the polyglucosan bodies of the brain, spinal cord and caecum. These results indicate that polyglucosan bodies contain mannose and glucose residues and suggest that the component of polyglucosan bodies is partially derived from rough endoplasmic reticulum, the Golgi apparatus and hypolemmal cisternae.

Reproduction of female Japanese serow based on the morphology of ovaries and fetuses

The Japanese serow (Capricornis crispus) is the only wild bovine ruminant preserved as a special natural monument in Japan. Until the present authors reported some data on the reproduction of males (Tiba et al. 1981a,b) and females (Sugimura et al. 1981, 1983, 1984, Kita et al. 1983a,b, 1986), there was little information in the literature regarding the reproduction of the serow (Asdell 1964). In the present study, the data previously reported and further information on the reproduction of female serows are presented based on morphological studies of the ovary and the fetus.

Immunohistochemistry and effect of vinblastine on intracisternal microtubules of the canine neurons

The immuno-histochemical and drug-binding properties of intracisternal microtubules (IMs) in the RER of canine neurons were studied with antitubulin IgG and vinblastine. Specific fluorescence against antitubulin IgG was not detected in the RER in which the IMs were aggregated but was observed in the cytoplasm outside the RER. Injection of vinblastine into the sympathetic ganglion did not change the size, shape or arrangement of IMs in the RER, although typical honeycomb crystals were formed near IM-containing RER. These findings show that the IM does not possess antigenicity to antitubulin IgG and has no vinblastine-binding site. Thus it is suggested that the IM may be a defective or modified microtubule.

Non-response of rough endoplasmic reticulum containing intracisternal microtubules to axotomy in the dog

Responses of the RER containing intracisternal microtubules in the canine ganglion nodosum to axotomy were observed at 3, 7, 14, 30, 50 and 99 days after section of the vago-sympathetic trunk. At 3 days, neurons showed no changes. From 7 days to 50 days, a retrograde reaction occurred in the cell bodies; nuclei were irregular in outline and eccentric in position and flattened RER disintegrated and dispersed. On the other hand, dilated RER containing intracisternal microtubules, which was detected as cytoplasmic inclusions under the light microscope, was present at the opposite pole to the eccentric nucleus. The ratio of areas of cytoplasmic inclusion to cell body significantly decreased. However, there were no changes in the size and arrangement of intracisternal microtubules within the RER. At 99 days, axotomized cell bodies had recovered and the RER containing intracisternal microtubules was indistinguishable from the contralateral controls. These findings indicate that the RER containing intracisternal microtubules is not disrupted by axotomy, though it is compactly packed at the periphery of cell bodies during the retrograde reaction. It appears, therefore, that the RER containing intracisternal microtubules does not respond to axotomy.

Concanavalin A binding sites of rough endoplasmic reticulum containing intracisternal microtubules in canine neurones

Lectin histochemistry was used to identify sugar residues of IM-containing RER in elderly canine sympathetic ganglionic neurones. IM-inclusions stained with ConA-peroxidase conjugate, but not with soybean agglutinin (SBA), wheat germ agglutinin (WGA), peanut agglutinin (PNA), Dolichos biflorus agglutinin (DBA), Ricinus communis agglutinin (RCA I) and Ulex europaeus agglutinin (UEA I). ConA-binding sites were visualized within cisternae of RER containing IM; reaction product was localized in IM-containing RER cisternae and IM. Inhibition with specific sugars (0.1 M alpha-methyl-D-mannoside or 0.5 M D-glucose) blocked the binding of ConA to IM-inclusions and normal Nissl substance. When a low sugar concentration (5 x 10(-3) M alpha-methyl-D-mannoside or 0.2 M D-glucose) was employed, IM-inclusions were still strongly ConA-positive, but normal Nissl substance was not. These results demonstrate that IM-containing RER have an excessive amount of carbohydrates (mannose or glucose-rich sugars) which are essentially detected in flattened RER under normal conditions and further indicate that glycoproteins in IM differ from those in cytoplasmic microtubules.