The morphology of the peripheral blood leukocytes of the roughtail gecko, Cyrtopodion scrabum, is carefully described in Wright-Giemsa and toluidine-blue-stained blood films, and in the living condition by phase-contrast microscopy, using supravitally stained preparations. Mature eosinophils, basophils and small lymphocytes commonly occur in the blood, while monocytes are rarely seen. In addition, macrophages are occasionally encountered, but neutrophils cannot be observed. Developmental stages in eosinophil and basophil differentiation can be seen. This study serves as a basis for the cytochemical localization of substances within these blood cells.

The morphology of the peripheral blood leukocytes of Hemidactylus turcicus is examined and carefully described in Wright-Giemsa stained blood films, and in the living condition by phase-contrast microscopy, employing supravitally-stained preparations. Mature eosinophils, basophils, and small lymphocytes occur in the blood. In addition, developmental stages in eosinophil differentiation and basophilic myelocytes and metamyelocytes are sometimes seen. Medium lymphocytes are occasionally observed. Neutrophils are not seen; very rarely are monocytes observed. This study provides a basis for the cytochemical study of these cells which has its foundation in morphology.

The morphology and cytochemistry of the blood basophil of Bufo marinus are characterized, and compared and contrasted to the basophil from several other amphibians and with the mammalian basophil. The basophil of B. marinus contains at least one type of mucopolysaccharide and an acid mu- copolysaccharide, possibly heparin, within the cytoplasmic granules. Several amino acids, but no histidine, and no lipid, occur in B. marinus basophils. Furthermore, these basophils appear devoid of hydrolytic enzymes, excepting nonspecific esterase. They do possess numerous oxidative enzymes, however, in the intergranular cytoplasm. The basophil of B. marinus is more similar cytochemically to the mammalian basophil than to the basophil of several previously studied amphibians. Considerable information regarding the morphology and cytochemistry of the mammalian blood basophil is available (Wetzel et al., 1967; Ackerman and Clark, 1971; Komiyama and Spicer, 1974; van El- ven et al., 1977). Ackerman (1963a) pre- sents an excellent review and comparison of basophil cytochemistry in several mam- malian species, including man; he also compares and contrasts the basophil with the mast cell. In contrast, although discov- ered over a century ago, the precise func- tions of the mammalian basophil remain obscure, despite its involvement to some extent in phagocytosis, allergic reactions, and delayed hypersensitivity (Stossel, 1977). Even less is known about the am- phibian basophil. Due to the fragility of the basophil, and its infrequency (1% or less) in the peripheral blood (factors which also hindered study of this cell in mam- mals) few data regarding the amphibian basophil are available. Caxton-Martins (1978) cytochemically examined the leu- kocytes in two West African anurans, Rana temporaris and Bufo regularis, but reported his findings as granulocytes, not differ- entiating among neutrophils, eosinophils, and basophils. In the newt, Notophthalmus viridescens, Cowden et al. (1964) demon- strated the presence of mucopolysaccha- ride and tyrosine in basophilic leukocytes and tissue mast cells.

The integument of Bufo marinus is surveyed. It is similar to that of other anurans and contains the three chromatophores common to anurans. The skin of these toads contains more than 28% minerals deposited as small crystals in a mucopolysaccharide‐positive amorphous layer between the stratum compactum and the stratum spongiosum. These crystals reveal a high content of calcium and phosphorus aggregated in and near membrane‐bound vesicles which have an appearance very similar to matrix vesicles. Electron microscopy gives the appearance that these vesicles are associated with flbroblasts located between alternating bundles of collagen. Histochemical studies indicate that the amorphous layer possesses many of the characteristics commonly associated with mineralized cartilage or bone.

Neuronal perikarya and arterioles of slices of mouse midbrain were examined histochemically to determine their metabolic profiles. No differences in reactivities of key metabolic enzymes were observed between fresh 400-micron tissue sections and sections undergoing in vitro incubation for 4 h at 35 degrees C. Both neurons and arterioles appear capable of aerobic and anaerobic metabolism, while fatty acid utilization is limited. An operative hexose-monophosphate shunt occurs in midbrain neurons and arterioles. These data strongly suggest that electrophysiological and neurochemical studies using the in vitro preparation yield similar data to those obtained from fresh tissue.

Blood leukocytes of Bufo alvarius were studied by light and phase-contrast microscopy and histochemical techniques for the localization of glycogen and several hydrolytic enzymes, i.e., acid and alkaline phosphatases, nonspecific esterase, beta-glucuronidase, aryl-sulfatase, and myeloperoxidase (peroxidase). Neutrophils were the only leukocytes to demonstrate alkaline phosphatase activity, while beta-glucuronidase and aryl-sulfatase were not observed in any leukocytes. Periodic acid – Schiff (PAS) positive granules and granules containing hydrolytic enzymes occurred in varying amounts in leukocytes. In eosinophils, most glycogen was associated with smaller granules, while the larger refractile granules were PAS negative. Small lymphocytes were myeloperoxidase (peroxidase) negative. The present study agrees with previous investigations in mammals which indicate that specific granules in granulocytes may be PAS positive as well as contain one or more hydrolytic enzymes. In small lymphocytes of B. alvarius, PAS positive and acid phosphatase positive granules correspond to neutral red granules seen in supravital films. Furthermore, the appearance and histochemical reactivity of acid phosphatase granules in mature neutrophils, metamyelocytes, and late myelocytes correspond closely with the appearance and number of specific neutrophilic granules seen in Wright–Giemsa preparations and with PAS positive granules.

No differences in arteriolar metabolic profiles from rat caudate nucleus, ventral tegmental area or substantia nigra were observed between saline-control and methamphetamine HCl-treated (20 mg/kg, i.p. twice daily for 10 consecutive days) animals. Arterioles from the above regions of forebrain and midbrain are metabolically active vessels with a capacity for aerobic and anaerobic metabolism. These results suggest that a high dose of methamphetamine does not alter cerebral arteriolar metabolism in the areas examined.

To determine the metabolic profiles of arterioles of the rat ventral tegmental area and zona compacta and zona reticulata of the substantia nigra (SN), the distribution of selected enzymes, or by-products, of key metabolic pathways were examined histologically. Arterioles of all three regions expressed the enzymes required for aerobic and anaerobic metabolism. However, the relative abundance of the enzymes and byproducts suggests a lower metabolic capacity for the SN than the ventral tegmentum, while lipid catabolism in both regions appears non-operative. Moreover, the larger ventral tegmental arterioles possess a greater potential for nucleic acid and protein synthesis. Together, these results suggest the larger ventral tegmental arterioles possess a greater capacity for proliferation and repair.

Using immunohistochemical methods with antibodies specific to tyrosine hydroxylase, we examined the distribution of dopaminergic cells in the dorsal and median raphe nucleus of the rat brain. Although dopamine-containing cell bodies were previously thought to be almost exclusively confined to the substantia nigra pars compacta, ventral tegmental area, and tuberoinfundibular system, we found numerous cell bodies which stained for tyrosine hydroxylase in the dorsal and median raphe nuclei.

Journal Article Dr. Black and the "Amalgam Question" Get access M. SAMUEL CANNON, M. SAMUEL CANNON Department of Anatomy, College of Medicine Texas A&M UniversityCollege Station, Texas 77843-1114 Search for other works by this author on: Oxford Academic PubMed Google Scholar EVELYN D. KAPES, EVELYN D. KAPES Department of Anatomy, College of Medicine Texas A&M UniversityCollege Station, Texas 77843-1114 Search for other works by this author on: Oxford Academic PubMed Google Scholar GABRIEL A. PALKUTI GABRIEL A. PALKUTI Department of Anatomy, College of Medicine Texas A&M UniversityCollege Station, Texas 77843-1114 Search for other works by this author on: Oxford Academic PubMed Google Scholar Journal of the History of Medicine and Allied Sciences, Volume 40, Issue 3, July 1985, Pages 309–326, https://doi.org/10.1093/jhmas/40.3.309 Published: 01 July 1985

Chronic administration of methamphetamine (20 mg/kg, IP, every 12 hours for 10 days) produced a large decrease in tyrosine hydroxylase staining axons and terminal boutons in the caudate nucleus in rats when examined 60 days following the final methamphetamine injection. This effect was quantitated using the Leitz Data Acquisition and Display System (DADS) revealing that there was a 74% decrease in tyrosine hydroxylase positive processes in the caudate nucleus. Furthermore, this treatment also produced a large decrease in the number of tyrosine hydroxylase positive staining neuronal perikarya in the pars compacta of the substantia nigra. This effect was also quantitative using the Leitz-(DADS) system, revealing a decrease of 89% in tyrosine hydroxylase positive material. These data demonstrate that chronic administration of methamphetamine produces a long-term loss of tyrosine hydroxylase enzyme in both the cell bodies of the substantia nigra and the nerve terminals in the caudate nucleus. Whether this effect is due to the degeneration of the neurons or some metabolic effect remains to be determined.

Arterioles of the rat caudate nucleus were examined histochemically to determine their metabolic profile. These microvessels appear capable of aerobic and anaerobic metabolism with a potential for nucleic acid and protein synthesis. Little intramural lipid storage occurs and any fatty acids utilized are provided via the blood supply. Likewise, glycogen is not seen in the arteriolar wall and may be rapidly turned over as a substrate for anaerobic metabolism.

Coronary arteries and arterioles from normal rats, from rats made hyperthyroid by administration of desiccated thyroid for 10 weeks, and from hyperthyroid rats which were then fed normal control diets for 10 weeks, were examined histochemically to determine the activity of key metabolic pathways. The primary aims of this study were to determine if the alterations in particular enzyme and substrate activities that occur in thyrotoxic rat myocardium, arteries and arterioles were reversible and would return to normal levels following cessation of the hyperthyroid state. Our results suggest that hyperthyroid rats, even after 10 weeks on the normal diet, still show some compromise in arteriolar aerobic metabolism in favor of anaerobic pathways, while coronary arteries still demonstrate little glucose-6-phosphate dehydrogenase activity. Myocardial metabolic activity approximates that of normal control animals by the end of the 10th week on the normal diet.

A histochemical study of the metabolism of rat renal arteries and arterioles. Rat renal arteries and arterioles were examined histochemically to determine their metabolic profiles. Succinate, malate and NAD-isocitrate dehydrogenase, cytochrome oxidase and ubiquinone were assessed to determine aerobic metabolism. Glucose-6-phosphate dehydrogenase and DPN diaphorase were evaluated to determine hexosemonophosphate-shunt activity. Anaerobic metabolism was evaluated via lactate dehydrogenase, and the substrate, glycogen. Gomori's lipase, β-hydroxybutyrate dehydrogenase and amounts of neutral fat and free fatty acids were assessed as indicators of lipid utilization. Myosin ATPase activity was evaluated as an index of ATP utilization for contraction. Deoxyribonucleic and ribonucleic acids were appraised as indicators of protein synthesis. In general, the oxidative enzymes and myosin ATPase demonstrate considerable activity in renal arteries and arterioles which suggests aerobic metabolism and ATP usage. Renal arteries and arterioles also appear capable of anaerobic metabolism as indicated by strong lactate dehydrogenase reactivity and by the presence of slight to moderate quantities of glycogen, while high levels of glucose-6-phosphate dehydrogenase and moderate amounts of deoxyribonucleic acid suggest a potential for nucleic acid and protein synthesis. In arteries and arterioles, strong reactivity for β-hydroxybutyrate dehydrogenase, minimal lipase activity, and the absence of fatty acids with substantial amounts of neutral fat, indicate limited lipid catabolism.

Recording sites from single unit electrophysiological studies in vitro can be precisely localized by first marking the recording locus either by depositing Fast Green dye (for micropipette studies) or electrolytic lesioning (for metal electrode studies). The slices are then fixed in paraformaldehyde, placed in sucrose and attached to a coverslip by the surface tension of water. The slices are attached to a base brain in a cryostat so that the sections can be cut at the proper angle. The slices are then stained using a Nissl staining protocol. This procedure provides intact sections from small tissue slices with the recording locus clearly demarcated.

Spinal cord arteries and arterioles of adult female rats were examined histochemically to determine their metabolic profiles. The metabolic pathways evaluated included those related to aerobic (oxidative phosphorylation, Kreb's cycle and respiratory chain) and anaerobic (glycolysis) capacity, hexosemonophosphate-shunt activity, beta-oxidation of fat and adenosine triphosphate utilization. The amounts of deoxyribonucleic and ribonucleic acids were determined as an indication of protein synthesis. The present findings indicate that arteries of the rat spinal cord are metabolically active with high capacities for both aerobic and anaerobic metabolism, and possess a significant potential for nucleic acid and protein synthesis. Lipid catabolism, via beta-oxidation of fat, may serve as one source of energy. The arteries also demonstrate a high capacity for utilization of adenosine triphosphate. In contrast, the spinal cord arterioles show a lower capacity for aerobic metabolism and lipid utilization, while anaerobic glycolysis may be a main source of energy. The arterioles also demonstrate a significant potential for nucleic acid and protein synthesis, in addition to a high capacity for adenosine triphosphate utilization.

The new electron carrier Meldola Blue was employed in the cytochemical demonstration of succinate, lactate, and glucose-6-phosphate dehydrogenases in leukocytes. A significant increase in dehydrogenase activity, with better localization and less diffusion of the reaction product, was observed when compared to enzyme reactivity carried out without Meldola Blue. Meldola Blue can facilitate the demonstration of both bound and soluble leukocyte dehydrogenases. Current methods which could yield better visualization of leukocyte dehydrogenases, particularly succinate, lactate, and glucose-6-phosphate dehydrogenases in neutrophils, suggest excellent use can be made of Meldola Blue in the clinical laboratory. Reliable histochemical determination of these enzymes can aid in differentiating younger from older neutrophils. In addition, glucose-6-phosphate dehydrogenase might be used in conjunction with the nitroblue tetrazolium (NBT) test in separating persons with bacterial infections from those with nonbacterial illness. Furthermore, histochemical visualization of this enzyme may also prove beneficial in identifying glucose-6-phosphate dehydrogenase-deficient individuals.

Acinar cells of extraorbital lacrimal glands from control, pilocarpine-treated, atropine-treated and atropine + pilocarpine-treated rats were studied using a potassium pyroantimonate technique and X-ray microanalysis for calcium localization at the ultrastructural level. This was done in order to identify intracellular compartmentalization of calcium and to elucidate any calcium translocation that might occur during the secretory process. Calcium-pyroantimonate complexes were identified in the mitochondria, plasma membrane and cytoplasmic vesicles of the untreated specimens and in the plasma membrane of atropine-treated specimens, these complexes decreased drastically in the actively-secreting cells. The function of calcium in lacrimal gland secretion and the action of pilocarpine and atropine on membrane calcium are discussed.

Coronary arteries and arterioles in the left ventricle from the primate Macaca fascicularis were histochemically examined to evaluate their metabolic profiles. Succinate dehydrogenase and cytochrome oxidase activities were assessed to evaluate aerobic metabolic capacity, while myosin ATPase activity was determined as an index of ATP utilization for contraction. Anaerobic capacity was evaluated from lactate dehydrogenase and glycogen reactivity. Glucose-6-phosphate dehydrogenase was examined to determine capacity of the hexose-monophosphate-shunt, while the amounts of deoxyribonucleicc and ribonuclei acids were assessed as possible indicators of protein synthesis. Succinate dehydrogenase and cytochrome oxidase demonstrated slight reactivity in both coronary arteries and arterioles indicating a low capacity for aerobic metabolism. Myosin ATPase showed strong activity in arteries and even stronger reactivity in arterioles, suggesting that arteriolar smooth muscle is more capable of utilizing ATP. Glucose-6-phosphate dehydrogenase activity was extremely low in both arteries and arterioles, while deoxyribonucleic and ribonucleic acids demonstrated only slight to moderate reactivity in both arteries and arterioles, indicating that under normal conditions the coronary vasculature appears quite stable with little cell proliferation.