Quantitative enzyme histochemistry of rat foetal brain and trigeminal ganglion

Summary The increasing concern and the efforts in determining neurological effects in offsprings resulting from maternal exposure to xenobiotics are faced with several difficulties in monitoring damage to the central nervous system. In this paper, the efficiency of several enzyme histochemical reactions for analysing the forebrain and the trigeminal ganglia of rat foetuses are reported. Brains of 20-day-old Sprague-Dawley rat foetuses were frozen and analysed for 18 enzymes that had previously been used to monitor initial injury caused by toxic compounds in liver and other organs. Eight enzymes appeared suitable as histochemical markers for the functional integrity of different areas in brain and ganglia of rats exposed to xenobiotics. They were lactate, malate, glycerophosphate (NAD-linked), succinate, aldehyde and glucose 6-phosphate dehydrogenases, -glycerophosphate-menadione oxidoreductase and cytochromec oxidase. The activities of the enzymes were determined by microphotometry and the arrangement of absorbances of the enzyme final reaction products into appropriate analytical tables is proposed as an efficient procedure for data analysis.Abbreviations AcChE acetylcholinesterase - AldDH aldehyde dehydrogenase - ALKPase alkaline phosphatase - 5AMPase adenosine monophosphatase - ATPase Mg²⁺ dependent adenosine triphosphatase - CytOx cytochromec oxidase - GAPDH glyceraldehyde phosphate dehydrogenase - GIDH glutamate dehydrogenase - GLPDH glycerophosphate: NAD oxidoreductase - CPODH glycerophosphate:menadione oxidoreductase - G6Pase glucose-6-phosphatase - G6PDH glucose-6-phosphate dehydrogenase - IDH lactate dehydrogenase - MaDH malate dehydrogenase - MAO monoamine oxidase - NADPH, DH, NADPH tetrazolium oxidoreductase - SuDH succinate dehydrogenase - 6PGDH 6-phosphogluconate dehydrogenase     

Light optical and electron microscopic study of the nodose ganglion neurons in emotional stress

Cell bodies of cardiovascular receptors localized in the ganglion nodosum of rabbits exposed to experimental emotional stress were studied with the light and electron microscope. Two groups of animals were selected for investigation. Under emotional stress rabbits of one group demonstrated almost unchanged arterial pressure and only a small increase in heart rate, while animals of the other group displayed strongly marked disturbances of their blood circulation leading to the lethal outcome at the end of experiment. In the first group rabbits, the microscopic anatomy and ultrastructure of the nodose ganglion neurons indicated an increased activity in the nerve cell. At the same time, morphological evidences of exhaustion were revealed in neurons of the nodose ganglion of the second group rabbits. A possible role of the distortion of the afferent information in pathogenesis of cardiovascular disorders under emotional stress is discussed.     

Light and electron microscopic histochemical observations on cholinesterase-containing sympathetic nerve fibres in the pineal body of the rat

Synopsis Pineal glands of adult albino rats were examined histochemically using, first, formaldehyde-induced fluorescence to study monoamines and, second, copper thiocholine or copper ferrocyanide methods to study acetylcholinesterase and non-specific cholinesterase by light and electron microscopy. Cholinesterase was determined quantitatively by a constant pH titration assay.Fluorescent and acetylcholinesterase-positive nerve nets formed identical patterns. Nonspecific cholinesterase was observed only in nerve trunks outside the pineal. Bilateral removal of superior cervical ganglia resulted in complete disappearance of fluorescence and acetylcholinesterase from nerve fibres. Electron microscopically, acetylcholinesterase was found on sympathetic axons containing small granular vesicles. Quantitative cholinesterase determinations suggested that the pineal activity was mainly due to acetylcholinesterase. Comparison of the incubation times required for equal histochemical acetylcholinesterase reactions suggested that the activity of the sympathetic nerve fibres in the pineal is of the same order of magnitude as that in the nerve fibres of the iris.     

Acid phosphatase of osteoclasts demonstrated by electron microscopic histochemistry

Summary Location of acid phosphatase outside and inside the osteoclast was studied by electron microscopic histochemistry. Osteoclasts with a ruffled border apposed to the bone showed enzyme activity in a) membrane-limited cytoplasmic bodies of different dimensions, b) some Golgi vesicles and inner Golgi cisternae, c) vacuoles and vacuole-like profiles, d) extracellular channels and channel expansions in the ruffled border, e) cell-bone interspace. The possibility of bone degradation by lysosomal enzymes both in the cytoplasmic vacuoles and in the cell-bone interspace is discussed.This research was supported by the Danish Medical Research Council. Grant. no. 512-819.I am indebted to Professor Arvid B. Maunsbach for valuable discussions and suggestions and to Mrs. Ruth Nielsen for technical assistance.     

Light and electron microscopic localization of substance P-like immunoreactivity in the cerebral ganglion of locust with a monoclonal antibody

Summary The occurrence of substance P-like immunoreactivity was studied in the locust brain at light and electron microscopic level using monoclonal IgG fraction to substance P. Small immunoreactive perikarya have been found beside the medial neurosecretory cells in horizontal brain sections. Widespread immunoreactivity was also observed in the protocerebral neuropil notably in the central body and bordering on the corpora pedunculata. The reaction endproduct appeared as fine, more or less round particles in the central body, and as coarse varicosities and wavy fibres bordering the peduncles. The roundish particles probably represent nerve terminals, while the wavy fibers correspond to neural processes. In the vicinity of the lobe immunoreactivity was not observed. Electron microscopically, a number of immunoreactive terminals were found in the protocerebral neuropil. The reaction endproduct was accumulated mostly in large dense core granules/average diameter 80 nm/however reaction endproduct was also observed on the external surface membranes of clear vesicles and mitochondria. Our results suggest the widespread occurrence of a substance-P immunoreactive neuropeptide in the cerebral ganglia of the migratory locust.     

The incubation of unfixed tissues for electron microscopic histochemistry

Summary Small tissue blocks of various organs of the rat were incubated for gradually increasing times in a neutral buffer-sucrose medium modelling the main parameters of the histochemical incubations. Following incubation the blocks were fixed in paraformaldehydeosmium and embedded in Durcupan. The electron microscopic study revealed that even after 40 minutes of incubation prior to fixation, fine structure is preserved satisfactorily from the histochemical point of view. With consideration to further advantages of such a proceeding, the incubation of unfixed tissue blocks for electron-histochemistry is recommended.     

Light and electron microscopic studies of the ventricular wall

Summary Electron microscopic data confirm the results gained with rapid Golgi preparations of adult rodent brains that tanycytes occur in clusters along the lateral wall of the third ventricle. The cytoplasmic matrix of these cells is considerably denser than that of typical ependymal cells. They have filaments and microtubules throughout their cytoplasm along with mitochondria and polysomes. At the surface is a compact group of microvilli which suggest that tanycytes might selectively absorb material from the ventricle.The tanycytes are segregated from neuropil by other tanycyte processes, by neighboring ependymal cells and by astrocytes. Yet there are gaps in this sheath. At these points tanycytes either abut upon or surround nonglial components of the neural fabric.Their cytological features and relations with the neuropil suggest that tanycytes selectively absorb material from the ventricle and release it along the basal process, primarily affecting those segments of neurons immediately adjacent to the tanycyte.Supported by: NINDS Grants 5 R01 NS 09001-02 NEUA, 5T01 NB 5309, and GM 00958, and by the Eleanor Roosevelt Cancer Foundation Research Institute.Acknowledgements: This work was initiated in the Anatomy Department of the Harvard Medical School with facilities provided by Prof. S. L. Palay (U.S. Public Health Service Grant No. NB 05591). Dr. R. B. Wuerker kindly and patiently provided the instruction and orientation to electron microscopy. The major portion of the study was completed in the Neurology Department of the University of Utah with the extremely competent, challenging assistance of Dee Lerdahl, Nina Belgarian, Keith Johnson and Lynn Kendricks.     

Light and electron microscopic study of the mitochondria in human embryonic nervous tissue cells in vitro

After a 5-min heating of isolated Tradescantia leaves at 49 degrees C the granular component in the nucleoli of parenchyma cells disappears and the nucleoli grow more compact and exclusively fibrillar in structure. Simultaneously, the ability of chloroplasts of these cells for phototaxis is completely inhibited. In leaves placed in a moist chamber at room temperature 48 h after heating a normal ultrastructure of nucleoli and the ability of chloroplasts to respond by phototaxis to change in illuminations are restored almost synchronously.     

Light and electron microscopic studies of crayfish hemocytes

Using light and electron microscopy, three hemocyte types are described in the hemolymph of the crayfish. The coagulocyte comprises 65% of the total hemocyte number and contains medium-sized cytoplasmic granules, abundant dilated rough endoplasmic reticulum, and a highly developed Golgi complex. It rapidly undergoes cytolysis in vitro and participates in coagulation by releasing the contents of its granules to the hemolymph. The granulocyte comprises 31% of the total hemocyte number and is capable of phagocytosis. It contains large, irregularly shaped cytoplasmic granules, a moderately developed Golgi complex, and moderate amounts of non-dilated rough endoplasmic reticulum. During coagulation in vitro, the cell attaches and spreads onto the substratum; this is followed by a slow intracellular granule breakdown and cytolysis. The amebocyte comprises 4% of the total hemocyte number and it is also capable of phagocytosis. It possesses small cytoplasmic granules, many vacuoles, a moderately developed Golgi complex, and large amounts of smooth endoplasmic reticulum. It is distinguished from the other two cell types by being stable and motile in vitro.     

VIP like immunoreactive nerves in human respiratory tract. Light and electron microscopic study

The present study provides light and electron microscopical evidence of Vasoactive Intestinal Peptide - (VIP) like immunoreactive nerves in human lower respiratory tract. Peroxidase antiperoxidase (PAP) technique was used to localize VIP-like immunoreactivity light microscopically and ultrastructurally. Under light microscopy, VIP-like immunoreactive nerves were observed in the smooth muscle layer of secondary bronchi to small bronchioli, and in bronchial glands. In addition, positive immunoreactive nervous network to VIP was found around nerve cell bodies in small microganglia. The bronchial epithelium of airway tract did not receive any VIP positive nerve fibers. Ultrastructurally VIP-like positive immunoreaction was localized in large granular vesicles ranging from 90 to 210 nm. Usually VIP-like positive immunoreactive nerve profiles contained several immunoreactive large vesicles (100-210). However, nerve profiles containing only a few positive large vesicles (80-150) were also observed. Under electron microscopy VIP-positive nerve profiles corresponded ultrastructurally to nerve profiles containing large granular vesicles observed in conventional electronmicroscopy. The present study provides new information about the innervation of human lower airway tract and widens the concept of their functional regulation on the anatomical basis reported here.