Histochemical studies on the morphology of the Golgi apparatus and on the distribution of some enzymes concerned with carbodydrate metabolism in the rat cerebellum.

Detailed histochemical studies have been conducted on the distribution of various enzymes, including thiamine pyrophosphatase, alpha-glucan phosphorylase, hexokinase, glucose-6-phosphate dehydrogenase, aldolase, glycerol-3-phosphate dehydrogenase; menadion oxidoreductase, lactate dehydrogenase and succinate dehydrogenase in various components of the cerebellum of healthy adult male rats of the Wistar strain. The thiamine pyrophosphatase reaction showed the morphological patterns of the GOLGI apparatus characteristic for each kind of cells. The GOLGI apparatus is a simple network in stellate cells, but it can be classified into the same 5 categories in basket cells and GOLGI type II cells. The GOLGI apparatus in the latter 2 cell types appears to undergo cyclic changes. A few GOLGI type II cells have a supranuclear form (Type II) and some cells show disintegration and "budding-off" of the GOLGI apparatus. The GOLGI apparatus in PURKINJE cells can be classified into 4 categories including a perinuclear strand form (Type III), but most of them show randomly distributed granules and vesicles. Lightly stained networks are observable in astrocytes and oligodendrocytes. They do not show polarity in astrocytes whereas they have extensions in a few oligodendrocytes. BERGMANN glia may undergo cyclic changes indicating more advance differentiation than astrocytes and oligodendrocytes. Cerebellar glomerula show lightly stained networks with many fine granules. Granule cells, stellate cells, and basket cells are all poorly equipped equally with the EMBDEN-MEYERHOF (EM) pathway and with the hexosemonophosphate (HMP) shunt. GOLGI type II cells are richly equipped almost equally with both the EM pathway and the HMP shunt. All these neurons probably derive energy mainly from glucose in the circulating blood. PURKINJE cells may belong to the category of "usual neurons", because they are moderately equipped both with the EM pathway and the HMP shunt. However, they may derive their energy from the BERGMANN glia which have intense hexokinase activity but weak succinate dehydrogenase activity. The BERGMANN glia are more richly equipped with the HMP shunt than with the EM pathway and are rich in lactate dehydrogenase suggesting an "exceptional metabolic pattern". These glia may have active synthesizing ability. Astrocytes and oligodendrocytes are equipped with all the enzymes tested, and they show a tendency to surround the glomeruli. It is suggested that the glomerula may be surrounded by the glial sheaths with strong hexokinase activity, and that they may contain alpha-glucan phosphorylase, glucose-6-phosphate dehydrogenase, and glycerol-3-phosphate dehydrogenase in addition to the succinate dehydrogenase already reported. A few PURKINJE cells showed perinuclear concentrations of the reaction product only of succinate dehydrogenase at the sites of contacts between nucleoli and nuclear membranes. It is suggested that the nucleolus may receive adenosine at the sites of contacts between nucleoli and nuclear membranes...     

Histochemical studies on the distribution of hexokinase and several enzymes related to catecholamine production in the locus coeruleus of the squirrel monkey.

Protein sulfhydryl groups are histochemically demonstrated by reacting with DDD followed by coupling with Fast blue B. The molar absorptivity of the formed azo dye is 19000 per mole SH reacted. DDD simultaneously reacts with protein-SH- and -SS-groups. However, the reaction with SH-groups is approximately 1000 times faster than that observed with SS-groups. With Ehrlich ascites tumour cells the reaction of DDD with SH-groups is completed within 7 h while the reaction of DDD with SS-groups needs 14 days for completion. Due to the extreme difference in the reaction rates protein bound SH-groups as well as reactive SS-groups can be estimated quantitatively by cytospectrophotometrical methods. The cells investigated showed an average SH-content of (1,30 +/- 0,03) X 10(-14) M SH/cell while the average content of reactive SS-groups was (1,59 +/- 0,28) X 10(-14) M SS/cell. In addition it was found that especially the amount of reactive SS-groups per cell is not constant but exhibits seasonal variations.     

Histochemical studies on the morphology of the Golgi apparatus and its relation to catecholamine biosynthesis in the locus coeruleus of Rhesus and crab-eating monkeys.

Detailed histochemical studies have been conducted on the distribution of thiamine pyrophosphatase (TPPase), L-gulonolactone oxidase (GLO), and NAD-linked xylitol dehydrogenase (XDH) in every component of the Locus coeruleus (LC) of healthy adult male Rhesus and crab-eating monkeys in order to clearify the morphology of the Golgi apparatus (GA) and its relation to biosynthesis of catechol-amines in this special nucleus of the primate (NA nucleus A 6 as defined in the Rhesus monkey by German and Bowden [1975]). Medium-sized neurons of both species of monkeys, which are considered to play an important role in the LC, were classified into 5 groups on the basis of morphological patterns of the GA. Many neurons of both species of monkeys were positive for the XDH test while some neurons of the crab-eating monkey as well as a few neurons of the Rhesus monkey were positive for the GLO reaction. The LC of both species of monkeys must be composed of metabolically one kind of identical medium-sized parasympathetic neurons whose GA may continously undergo 5 distinct phasic changes depending on the functional state of that cell. However, the GA changes its shape much more significantly even within each group of the 5 in the crab-eating monkey than in the Rhesus monkey. The GA Type IV may correspond to the catabolic phase of the GA during which biosynthesis of both catecholamines and vitamin C should be going on. Production of vitamin C may greatly help biosynthesis of catecholamines in LC. The difference in species is evident between the 2 kinds of monkeys studied in regard to the degree of their ability to synthesize these substances. The degree of the ability to synthesize vitamin C parallels the density distribution of Type IV neurons in LC whose GA often develops much more greatly in the crab-eating monkey than in the Rhesus monkey.     

镉对慢性镉中毒小鼠黑质神经元的损害及维生素C的拮抗作用

目的研究维生素C（VC）对慢性镉中毒小鼠黑质神经元的保护作用。方法将75只雄性昆明种小鼠随机分为3组,每组25只。镉中毒组皮下注射氯化镉（CdCI：）,每周2次,每次2mg／kg,VC加镉组（VC组）上述方法和剂量染镉外,VC按每次25mg稀释后灌胃,正常组同时给予生理盐水。3个月后对小鼠进行Y型迷宫学习能力检测,并通过光镜及透射电镜观察小鼠黑质神经元的细微结构。结果Y型迷宫检测显示,VC组与镉中毒组相比,小鼠的学习、记忆能力均降低,差异有统计学意义（P〈0．05）;与正常组比较,差异无统计学意义（P〉0．05）。光镜和电镜结果显示,VC组小鼠中脑黑质区结构接近正常组,与镉中毒组比较有明显差异。结论抗氧化剂VC对慢性镉中毒组小鼠黑质神经元有显著的保护作用。     

Topographical distribution of substance P in the cat substantia nigra.

The substance P (SP) content in cat substantia nigra (SN) was estimated by radioimmunoassay in microdiscs punched on successive frontal sections. Only slight differences were seen in the regional distribution of SP within the SN. The highest content was found in the pars reticulata (20.7 pmol/mg protein) which was two times higher than in the pars compacta and pars lateralis. Identical results were found in the right and left part of the SN. Out of this area, as pedunculus cerebri and zona incerta, very low concentrations were detected.     

Histochemical studies on the relationship between the morphology of the Golgi apparatus and G6PD activity in the locus coeruleus and dorsal vagal nucleus of the rat (application of karyometry)

Histochemical studies on the distribution of TPPase and G6PD in the neurons of locus coeruleus (LC) and dorsal vagal nucleus (DVN) of rats have been conducted by using the TPPase method, the G6PD method, karyometry, and statistics under normal conditions. 1. Neurons of the LC and DVN may undergo phasic changes of the GOLGI apparatus (GA) under normal condition. 2. No significant difference was found between neurons of the LC and DVN in regard to the GA morphology, but in general the latter revealed more complicated and developed GA than in the former. 3. Surprisingly, the G6PD activity of LC neurons was very weak. This is in agreement with the previous report by IIJIMA and IMAI (1975), and in disagreement with FRIEDE (1966). 4. An intimate parallel relationship was found between the GA morphology and G6PD activity in the DVN. The findings support the vigorous synthesizing activity found in the previous study of the rat supraoptic nucleus (IIJIMA 1979). In contrast to the DVN, such a parallel relationship between the Ga shape and G6PD activity was absent in the LC. Both findings strongly suggest that the role of the GA may be functionally different in the LC and DVN.     

Transmitter synthesis increases in substantia nigra neurons of the aged mouse.

Striatal dopamine concentrations are relatively well maintained with age despite extensive death of the nigrostriatal neurons whose terminals contain the dopamine. Counts of nigrostriatal dopaminergic neurons in C57BL mice identified using immunocytochemistry, Fluoro-Gold retrograde axonal transport and Nissl staining were combined with measures of striatal dopamine and DOPA after saline, pargyline or NSD-1015 treatment. On average, 68% of the dopaminergic nigrostriatal neurons died between ages 8 and 104 weeks and there was a 3-fold increase in dopamine synthesis per average neuron in the aged mice. Increased transmitter synthesis by surviving neurons may serve to compensate brain function in old age.     

Immunocytochemical studies on the basal ganglia and substantia nigra in Parkinson's disease and Huntington's chorea

The basal ganglia and substantia nigra, taken from control human brain and from patients dying with a diagnosis of Parkinson's disease or Huntington's chorea, were analysed with histochemical and biochemical techniques. The pigmented neurons of the substantia nigra pars compacta possess tyrosine hydroxylase immunoreactivity and are disposed in three major layers, alpha, beta and gamma. This pattern became obscured in choreic brains by the severe shrinkage of the nigra, but total numbers of pigmented neurons were within the normal range. In contrast, pigmented neurons were lost from all layers of the substantia nigra in Parkinson's disease, although examination of cases with minimal cell loss suggested that an internal part of the lateral alpha sub-layer was most severely and consistently affected. A dopaminergic projection between this internal part of the alpha sub-layer and the putamen was suggested by the preferential loss of catecholamines from the putamen in Parkinson's disease. The distribution of the peptides, substance P, methionine-enkephalin and dynorphin 1-17 were mapped immunohistochemically within the substantia nigra. The different patterns of immunoreactive axons and terminals were found to be extensive, at least partially overlapping, and largely avoided the region of the pigmented perikarya of the alpha sub-layer and nucleus paranigralis. All peptides were depleted in choreic substantia nigra, reflecting the degeneration of the striatonigral pathway. However, concentrations of enkephalin-like immunoreactivity were increased within the interpeduncular nucleus. In Parkinson's disease there was a loss of enkephalin- and dynorphin-like immunoreactivity from the substantia nigra but a fall in substance P-like immunoreactivity was only detected by radioimmunoassay, not by immunocytochemistry. Peptide immunoreactivity was also reduced within choreic basal ganglia. However, no gross changes were found in peptide staining of the parkinsonian basal ganglia. In summary we have reported a number of changes in peptide-containing pathways in human degenerative disorders that may reflect the degeneration of neuronal pathways either as a primary event or secondary to initial lesion. We have also emphasized the sensitivity of the alpha sub-layer of nigral neurons to damage in Parkinson's disease. We suggest that the lower density of peptidergic fibres in the area of the perikarya may contribute to the susceptibility of these neurons to damage.     

Histochemical studies in stomatal apparatus of Phaseolus mungo Linn. I. Localization of enzymes and structural material.

In the present investigations, the localization of several enzymes (Acid Phosphatase, Peroxidase, succinic dehydrogenase, Phosphorylase, alkaline phosphatase, ATP-ase) and other substances in the guard and subsidiary cells as well as trichomes of the leaves of Phaseolus mungo, was carried out. Attempts were also made to follow the sequence of developmental stages starting with meristemoids and culminating in differentiated structures. The basic information thus obtained is used in interpreting the developmental physiology of stomatal differentiation as well as their cellular organisation. Histochemical observations made in the present studies are compared with the electron microscopical observations of Whatley (1972). It is proposed that mitochondria played a basic role in the functioning of the guard cells. The present studies also demonstrated activity of acid phosphatase in the guard cells and was localized in spherosomes. The latter varied in the activity for acid phosphatase and was dependent on the turgid level of the cell. Interestingly, enough localization of phosphatase could only be observed in spherosomes when the osmotic pressure in the cell was relatively low, once the osmotic pressure increased, the activity disappeared.