Localization of monoaminergic neurons in the central nervous system of Astacus astacus Linné (Crustacea)

Summary The cellular localization of biogenic monoamines in crustaceans was studied by means of a highly specific and sensitive fluorescence method devised by Falck and Hillarp. It was found that neurons displaying specific fluorescence in the central nervous system were confined to the protocerebrum, the medulla externa and interna and the ventral nerve cord. The method allows a distinction between the fluorophores of 5-hydroxytryptamine (and 5-hydroxytryptophan), which emit the yellow light, and the fluorophores deriving from the catecholamines (and DOPA), which emit the green light. Green-fluorescent neurons occurred abundantly in the aforementioned parts of the central nervous system while yellow-fluorescent neurons were sparsely present in the same parts.The present work has been carried out at the departments of Histology and Zoology at the University of Lund. The authors take great pleasure in expressing their warmest thanks for laboratory facilities, provided by Professors Erik Dahl (Zoological Institute) and Bengt Falck (Histological Institute).The research reported in this document has been sponsored by the Air Force Office of Scientific Research under Grant AF EOAR 66-14 through the European Office of Aerospace Research (OAR), United States Air Force and by a grant from the Swedish Natural Science Research Council 99-32 (nr 5995).     

Monoaminergic mechanisms in the nervous system of Lumbricus terrestris (L.)

Summary The localization and intraneuronal distribution of the monoaminergic transmitters in the nervous system of the earthworm, Lumbricus terrestris, have been investigated in detail with the aid of the histochemical fluorescence method of Falck and Hillarp.In the ventral nerve cord, many yellow fluorescent, 5-hydroxytryptamine containing neurons are found, but only few green fluorescent noradrenaline containing cell bodies, which, however, are numerous in the peripheral nervous system. There is an abundance of both fibre types in the neuropile.The 5-hydroxytryptaminergic neurons probably have a motor (possibly inhibitor) function; the adrenergic neurons in the body segments are supposed to have a receptor (exteroceptive and possibly proprioceptive) function.In the cerebral ganglion, both 5-hydroxytryptamine and noradrenaline containing neurons are found in large numbers, and there are closely packed numerous fibres of both types in the neuropile. Their function is more obscure, though an associative function can be presumed for some adrenergic neurons; smaller 5-hydroxytryptaminergic neurons might have a motor (perhaps inhibitor) function.Adrenergic sensory cells are found in the body integument, most frequently in the clitellum segments, in the prostomium, and in the roof of the buccal cavity. These cells give off varicose fibres that form a basi-epithelial network which is in communication with the green fluorescent sensory fascicles in the ventral nerve cord via the epidermal nerves, the ring nerves, and the segmental nerves. No direct adrenergic sensory-effector innervation of either circular and/or longitudinal musculature or gland cells seems to exist. No adrenergic free nerve endings in the body integument have been observed. Instead, there must be a synaptic contact with the motoneurons, either directly in the neuropile or via an interjacent neuron.No synaptic contacts have been observed in the ventral nerve cord between adrenergic or 5-hydroxytryptaminergic fibres and either the giant fibres or fluorescent or nonfluorescent perikarya.An adrenergic innervation of the pharynx musculature has been found, and sensory cells of a different type are present in and below the epithelium; here, a direct senso-motoric innervation of the pharyngeal musculature cannot be excluded. It is established that the adrenergic neurons in the stomatogastric nervous system have an exciting function on the pharynx, whereas a direct monoaminergic influence of the muscular movements of the intestine probably does not exist.Abbreviations Used A adrenaline - CA catecholamine - DA dopamine - 5-HT 5-hydroxytryptamine - MA monoamine - NA noradrenaline The research reported in this document has been sponsored by the Air Force Office of Scientific Research under Grant AF EOAR 67-15 through the European Office of Aerospace Research (OAR), United States Air Force, by the Swedish Natural Science Research Council (99-34, 6627), and by the Swedish Medical Research Council (B67-12X-712-02A).     

Remarkable adrenergic nerves in the exocrine pancreas

Summary The adrenergic nerves in the pancreas of mice, rats, guinea-pigs, rabbits, and cats were investigated with the fluorescence method of Falck and Hillarp. The relations between the adrenergic fibres and the vessels were studied by the injection of india ink into the vessels.Besides the normal manifestation of adrenergic fibres at the large vessels, some vessels of capillary size were also accompanied by adrenergic fibres. These fibres had a very weak fluorescence, and showed up regularly only when the animal had been treated with Nialamide and L-DOPA or dopamine to increase the catecholamine content of the adrenergic fibres. The weakness of the fluorescence is perhaps due to low transmitter concentration or to small size of the nerve fibres, or to both. A rough estimate indicated that either the transmitter concentration of the nerve fibre is at least approximately 100 times below that seen in adrenergic nerves in other tissues, or that the radius of the varicosities of the nerve fibres is less than 0.2 . Neither alternative has previously been recognized.The secretory acini of the pancreas seem to lack a direct adrenergic supply. In the intrapancreatic ganglia, non-fluorescent nerve cells were reached by adrenergic terminals. No adrenergic nerve cells were detected in the pancreas of rats and cats. Small intensely fluorescent catecholamine-containing cells were observed in connexion with the intrapancreatic ganglia of rats.The research reported in this document has been sponsored by the Air Force Office of Scientific Research under grant AF EOAR 67-15 through the European Office of Aerospace Research (OAR), United States Air Force, by the United States Public Health Service (grant NB 06701-01) by the Swedish Medical Research Council (project B 67-12X-712-02A), and by the Faculty of Medicine, University of Lund, Sweden.     

Adrenergic retinal neurons

Summary Adrenergic retinal neurons have been studied in cynomolgus monkeys, cats, rabbits, guinea-pigs, rats, and mice with the fluorescence technique of Falck and Hillarp. With some species variations, three adrenergic fibre layers have been observed: an outer adrenergic fibre layer (all species) at the border between the inner nuclear and inner plexiform layers, a middle adrenergic fibre layer (rabbits, guinea-pigs, rats, and mice) in the middle of the inner plexiform layer, and an inner adrenergic fibre layer (rabbits) at the border between the inner plexiform layer and the ganglion cell layer. Similarly, three kinds of adrenergic nerve cells have been found: a somewhat heterogenous group of outer adrenergic cells (all species) situated in the innermost cell rows of the inner nuclear layer, eremite cells (rabbits, guinea-pigs, rats, and mice) within the inner plexiform layer and alloganglionic cells (all species) with a position and appearance resembling some of the ordinary non-adrenergic cells of the ganglion cell layer. All the adrenergic cells are star-shaped with slender branching processes running to the different adrenergic layers.The research reported in this document has been sponsored by the Air Force Office of Scientific Research under grant AF EOAR 66-14 through the European Office of Aerospace Research (OAR), United States Air Force, by the United States Public Health Service (grant no. NB 05236-02), by the Swedish Medical Research Council (grant no. B 66-320), and by the Faculty of Medicine, University of Lund, Sweden.     

The pineal gland of the mole-rat (Spalax ehrenbergi,Nehring)

Summary A comparative investigation of the distribution of monoaminergic neurons in non-malacostracan crustaceans was performed with the histochemical fluorescence method of Falck-Hillarp.Two fluorophores were found: the more widespread of the two emits a green fluorescence; and the more sparsely distributed emits a yellow to brown-yellow fluorescence.Specific green fluorescent areas were shown to exist in the protocerebrum. The central body and the optic ganglia of the compound eye (where present) are always fluorescent. Moreover, the centre of the nauplius eye may have a green fluorophore, as in ostracods, and a neuropile area, here called the frontal area. These neuropile centres are known from ordinary histological studies of the nervous system. In addition, there are specific monoaminergic centres, such as the so-called dorsal area of phyllopods and anostracans as well as the copepod specific areas. Specific monoaminergic areas appear in the deutocerebrum and the suboesophageal ganglion where they are particularly well developed.Presumed sensory neurons in the cavity receptor organ of Artemia salina are shown to be monoaminergic. Monoaminergic sensory neurons have not been described previously in Arthropods.Presumed motor innervation of hind-gut and trunk muscles is also found, and it is concluded that in crustaceans neurons of every type (sensory, internuncial, motor) may be monoaminergic.We have enjoyed unrestricted laboratory facilities at the Department of Histology, Faculty of Medicine, and with great pleasure express our sincere thanks to Prof. Bengt Falck. — Grants from the Swedish Natural Science Research Council (2760-007), the Swedish Medical Research Council (04X-712), the Royal Swedish Academy of Science (Hierta-Retzius), the Royal Physiographic Society of Lund, and the University of Lund supported the work.     

The effect of castration upon the ultrastructure of the rat hypothalamus

Summary The arcuate complex, comprising the nucleus and the outer zone of the median eminence, was studied under the electron microscope in control and castrated rats of both sexes. One month after castration the arcuate neurons show signs of hyperactivity characterized by dilated cisternae of the endoplasmic reticulum, a large nucleolus, situated near the nuclear envelope and fewer granulated vesicles. The surrounding neuropile shows an increase in the number of granulated vesicles above the control level. Six months after castration the changes already described are more accentuated. In the outer zone of the median eminence the axons and terminals show a considerable increase in the number of granulated vesicles which is of the order of 50 per cent above the control. A correlation between the granulated vesicles and the high content in dopamine of the arcuate complex is postulated. The ultrastructural changes observed in the arcuate complex, after castration, are discussed in relation to the current knowledge on the histophysiology of this region of the hypothalamus and specially on the probable regulatory effect of monoamines on the secretion of gonadotrophins.Supported by grants from the Consejo Nacional de Investigaciones Cientificas y Técnicas and by the Air Force Office of Scientific Research (AF-AFOSR 963-67).We are deeply indebted to Mrs. Defilippi-Novoa and Mr. Alberto Saenz for their skilful assistence.     

Monoaminergic neurons in the nervous system of crustaceans

Summary Certain neurons in the nervous system of the malacostracan crustaceans give rise to a predominantly green and a sparse yellow fluorophore in the histochemical fluorescence method of Falck-Hillarp. The same applies to the whole of Crustacea. The green fluorophore is probably a catecholamine; the yellow to brown-yellow has not yet been identified.The biogenic amine responsible for the green fluorescence, besides being found in diffusely distributed fibres, also appears in distinct areas of fibre concentrations in the central nervous system. The protocerebrum of the malacostracans contains three areas: the central body and two areas in the top of the brain, one anterior and one posterior. The latter two are not recognized as separate areas in ordinary histological preparations. In addition, the optic neuropiles are fluorescent, some with a distinct stratification of the fluorophore. The deuto and tritocerebrum and the ventral nerve cord also contain monoaminergic neurons. Of the brightly fluorescent areas in the whole of Crustacea, only the central body consistently exists in all species. The other areas of concentrated fluorescent neuropile are restricted to smaller taxonomic units and differ from each other. p The monoaminergic neurons in Crustacea are sensory, motor, and internuncial, and also belong to a fourth type which mimics the neurosecretory neurons in neurohaemal organs. Only one example of a monoaminergic sensory neuron is known (in Anemia, a non-malacostracan, Aramant and Elofsson 1976), a few motor and a few neurosecretory mimics (the latter in malacostracans). Most are internuncials. Acknowledgement. We have enjoyed the laboratory facilities at the Department of Histology, Faculty of Medicine, and express our sincere thanks to Prof. Bengt Falck.-Grants from the Swedish Natural Science Research Council (2760-007) and the Swedish Medical Research Council (04X-712) supported the work     

Quantitative fluorescence studies of the effects of catecholamines and hydrocortisone on endogenous amine levels in neurones and small intensely fluorescent cells of embryonic chick sympathetic ganglia in vivo and in vitro

Summary Chick embryo lumbar sympathetic ganglia (11 day) cultured for three days and uncultured (in vivo) ganglia of comparable age were freeze-dried and processed by the formaldehyde-induced fluorescence technique for the demonstration of biogenic monoamines. The catecholamine levels within principal neurone cell bodies and small intensely fluorescent (SIF) cells were then examined in plastic sections of the in vivo and in vitro ganglia by a quantitative fluorescence method under various experimental conditions. Culture of ganglia for three days in the presence of hydrocortisone acetate (10g/ml) resulted in an increased SIF cell fluorescence (P<0.001 compared to control) and a green to yellow colour shift in the fluorophore of SIF cells. No detectable alteration in the fluorescence level of neurones was observed. When neurones after three days in culture were incubated for 1 h in exogenous catecholamines, a significant increase in fluorescence levels (interpreted as an increase in catecholamine content) occurred with noradrenaline (2×10^–6 M; 2×10^–5 M). SIF cells in ganglia removed directly from 14-day old chicks similarly took up noradrenaline and dopamine, and also adrenaline (2×10^–5 M). Morphological results are presented which indicate that the cellular appearances and architecture of cultured ganglion explants are very similar to those in comparable ganglia in vivo.This work was supported by a grant from the Medical Research Council. We thank Mrs. G. O'Shea, Mr. T.T. Lee and Mr. P.F. Hire for their valuable technical assistance     

Direct and decarboxylation-dependent effects of neurotransmitter precursors on firing of isolated monoaminergic neurons

To elucidate mechanisms that underlie the profound physiological effects of the monoamine precursors 5-hydroxy-l-tryptophan (5-HTP) and l-3,4-dihydroxyphenylalanine (l-DOPA), we examined their action on single monoaminergic neurons isolated from the ganglia of the gastropod snail Lymnaea stagnalis. In isolated serotonergic PeA motoneurons, 5-HTP produced excitation. The effect was mimicked by serotonin at 0.5–1 μM, masked by pretreatment with serotonin at higher concentrations, and abolished by the inhibitor of aromatic amino acid decarboxylase (AAAD) m-hydroxybenzylhydrazine (NSD-1015), the inhibitor of the vesicular monoamine transporter reserpine or the serotonin receptor antagonist mianserin. Exposure of the dopaminergic interneurons RPeD1 to l-DOPA caused a biphasic effect composed of a depolarization followed by a hyperpolarization. AAAD inactivation with NSD-1015, as well as the blockade of dopamine receptors with sulpiride, resulted in the enhancement of the excitatory effect, and the abolition of the inhibitory effect. Dopamine caused hyperpolarization and masked the inhibitory phase of l-DOPA action. The results show that precursors affect the rate of firing of isolated monoaminergic neurons and that their effect is completely or partially mediated by the enhanced synthesis of the respective neurotransmitter, followed by extrasynaptic release of the latter and activation of extrasynaptic autoreceptors.     

Ultrastructure of the peptidergic and monoaminergic neurons in the hypothalamic neurosecretory system of Anuran Batracians

Summary In the toad Bufo arenarum Hensel the following regions of the hypothalamic — neurohypophyseal system were studied under the electronmicroscope: preoptic and paraventricular nuclei, median eminence and infundibular process of the neurohypophysis.Neuronal perikarya of the preoptic nucleus are loaded with typical neurosecretory granules of peptidergic nature having a mean diameter of 1660 Å. While most neurons of the winter toad are in a storage stage a few show signs of a more active synthetic activity. A distinctive feature of preoptic neurons is the presence of large lipid droplets. The paraventricular nucleus contains small neurons containing granulated vesicles with a mean diameter of 800-1000 Å. In the region extending between these two nuclei and the median eminence axons containing either neurosecretory elementary granules or granulated vesicles are observed.The inner zone of the median eminence is occupied by axons of the preoptic neurohypophyseal tract; two types of axons, according to the size and density of the neurosecretory granules, may be recognized. The outer zone of the median eminence contains mainly axons and nerve terminals containing granulated vesicles of probable monoaminergic nature and only a few with granules of peptidergic type.The neurohypophysis contains two kinds of axons: one with more dense granules of 1800 Å and the other with granules of lesser electron density and 2100 Å. At the ending proper small clear vesicles of synaptic type are found.A progressive increase in volume of the peptidergic granules along the axon is demonstrated. This is of the order of 218% from the preoptic perikarya down to the infundibular process. The physiological significance of the two neurosecretory systems — i.e. the monoaminergic and the peptidergic — and the probable nature of the two types of peptidergic axons is discussed.Supported by grants from the Consejo Nacional de Investigaciones Científicas y Técnicas and by the Air Force Office of Scientific Research (AF-AFOSR 963-67).The authors want to express their gratitude to Mrs. Defilippi-Novoa and Mr. Alberto Sáenz for their skillful assistance.     

5-Hydroxytryptamin im Gehirn der Eidechse (Lacerta viridis und Lacerta muralis)

Zusammenfassung Mit Hilfe der Methode von Falck und Hillarp wurde die Verteilung von 5-Hydroxytryptamin im Zentralnervensystem von Lacerta viridis und muralis untersucht. Mikrospektrometrische Analysen zeigen, daß sich die Gelbfluoreszenz wie formaldehyd-kondensiertes 5-Hydroxytryptamin verhält; chemische Bestimmungen ergeben hohe Werte von 5-Hydroxytryptamin im Gehirn der Eidechsen (5,2–6,4 g/g). Kerngebiete des Zwischen-, Mittel und Vorderhirns, die überwiegend in somatosensible oder sensorische Bahnen eingeschaltet sind, werden von Endaufsplitterungen 5-hydroxytryptaminhaltiger Neurone erreicht. Es wird angenommen, daß das Ursprungsgebiet dieser Fasersysteme im Tegmentum liegt. Der Nucleus reticularis mesencephali enthält zahlreiche Nervenzellen, deren Perikaryen einen hohen Gehalt an 5-Hydroxytryptamin aufweisen.

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5-Hydroxytryptamine in the brain of Lacerta viridis and Lacerta muralis

Summary The distribution of 5-hydroxytryptamine in the central nervous system of the lizards Lacerta viridis and muralis was investigated with the fluorescence method of Falck and Hillarp. Microspectrometric analyses revealed that the yellow fluorescence had the characteristics of the fluorophore of 5-hydroxytryptamine and chemical determinations on whole brains demonstrated the presence of considerable quantities of 5-hydroxytryptamine (5,2–6,4 g/g). Nuclear areas of the mesencephalon, di and telencephalon, which are mainly intercalated in sensory pathways, receive terminal ramifications of 5-hydroxytryptaminecontaining neurons. These fibres are presumed to originate from cells situated in the tegmentum. The nucleus reticularis mesencephali is shown to contain numerous perikarya of nerve cells rich in 5-hydroxytryptamine.

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Mit dankenswerter Unterstützung durch die Deutsche Forschungsgemeinschaft und die Joachim Jungius-Gesellschaft zur Förderung der Wissenschaften, Hamburg.

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Supported by grants from the Swedish Natural Science Research Council/project no. 99-35, and the Swedish Medical Research Council/project no. B 68-12x-712-03B.     

A canonical form for neural nets without circles

By “neural net” will be meant “neural net without circles.” Every neural net effects a transformation from inputs (i.e., firing patterns of the input neurons) to outputs (firing patterns of the output neurons). Two neural nets will be calledequivalent if they effect the same transformation from inputs to outputs. A canonical form is found for neural nets with respect to equivalence; i.e., a class of neural nets is defined, no two of which are equivalent, and which contains a neural net equivalent to any given neural net. This research was supported by the U.S. Air Force under Contract AF 49(638)-414 monitored by the Air Force Office of Scientific Research.     

Monoamine-containing neurons in the optic ganglia of crustaceans and insects

Summary With the fluorescence method of Falck and Hillarp, the presence and localization of monoaminergic neurons in the optic ganglia of several crustaceans and insects have been investigated. It was found that in both classes the monoaminergic terminals, when present, appeared (especially in the medullae externa and interna of the crustaceans and the medulla of the insects) in strata specific for each species. So far, the only monoamine (visualized by this technique) present in the crustacean optic ganglia is dopamine, whereas in the Insecta, the catecholamines dopamine and noradrenaline, and the indolamine, 5-hydroxytryptamine, are found in the optic lobe. But in the Insecta, different species show different content of these amines.This work was supported by grants 2760-3 and 2760-4 from the Swedish Natural Science Research Council (R.E.), by a fellowship from Deutsche Forschungsgemeinschaft, and a grant from the Swedish Medical Research Council B72-14X-712-D7B (N.K.). We are very grateful to the director of the Department of Histology, Faculty of Medicine, Lund, Professor Bengt Falck, who put all his facilities and knowledge at our disposal.     

Fluorescence methods for the histochemical demonstration of monoamines

Summary The histochemical fluorescence method of Falck and Hillarp for the demonstration of catecholamines and certain tryptamines, e.g. 5-hydroxytryptamine is based on the principle that these amines can be condensed with formaldehyde to yield strongly fluorescent 6,7-dihydroxy-3,4-dihydroisoquinolines and 6-hydroxy-3,4-dihydro--carbolines respectively. The investigation here reported presents the fluorescence characteristics and relative fluorescence yields for formaldehyde treated biogenic monoamines and certain related compounds enclosed in a dried protein layer. The fluorescence properties of some synthetic 6,7-substituted-3,4-dihydroisoquinolines and 3,4-dihydro--carbolines are given, and the fluorescence characteristics in relation to the molecular structure are discussed.Abbreviations used A adrenaline - DA dopamine - DOPA 3,4-dihydroxyphenylalanine - DOPS 3,4-dihydroxyphenyl-serine - 5-HT 5-hydroxytryptamine - 5-HTP 5-hydroxytryptophan - 5-MT 5-methoxytryptamine - -m-DA -methyl-dopamine - -m-DOPA -methyl-3,4-dihydroxyphenylalanine - -m-NA -methyl-noradrenahne - MTA 3-methoxy-tyramine - NA noradrenaline - NM normetanephrine - T Tryptamine - Try Tryptophan     

Monoamines in the liquor-contacting nerve cells in the hypothalamus of the lamprey,Lampetra fluviatilis L.

Summary The hypothalamus of adult lampreys (Lampetra fluviatilis L.) was studied by means of light and fluorescence microscopy (Falck's technique). Some single liquorcontacting nerve cells (LCNC) showing a weak green fluorescence were demonstrated in the ventral part of the third ventricle, above the preoptic recess. Caudally numerous fluorescent LCNC occur in the ventral part of the third ventricle, in the infundibular and in the posterior recess. The LCNC are to be observed between or below the ependymal cells lining the ventricular wall. These cells appear to be of the bipolar type. One process with a club-like protrusion is directed into the ventricular lumen, the other one into the opposite direction. Two types of fluorescent LCNC were distinguished: yellowish green cells, containing catecholamines, and yellowish orange cells, containing 5-hydroxytryptamine. Some similarity between the hypothalamic monoaminergic LCNC in lampreys and LCNC of the paraventricular organ of the other vertebrates was found. The localization, structure and monoaminergic nature of the hypothalamic LCNC in lampreys suggest the possibility, that their monoamines are released into the cerebrospinal fluid.I am very obliged to Prof. A.L. Polenov for his continuous help and advice. The skilful technical assistance of Mrs. G.N. Yakshina is gratefully acknowledged.     

Electron microscopic observations on the reactional changes occurring in insect nerve fibers after transection

Summary An electron microscopic study of the interganglionic connectives of an orthopteran (Laplatacris dispar) demonstrated that a reaction starts in the sectioned fibers very soon after transection (30 minutes). This reaction is closely similar to that observed in sectioned nerves of vertebrates and consists in the appearance of microvesicles and the proliferation of mitochondria. Sectioned connectives were studied from 30 minutes to 88 hours after section. The reaction mentioned increases progressively during the indicated lapse of time.This research was supported by the United States Air Force under Contract No AF 49 (638) 585 and Grant Af 61-64, monitored by the Air Force Office of Scientific Research, Air Force Research Division of the Air Research and Development Command.     

Decision equation for binary systems. Application to neuronal behavior

Summary The exact mathematical treatment is given for a non linear equation describing the delayed yes-or-no response to a binary system to external stimulations, in some typical cases of interest. Comparison is made with neurophysiological data on the frequency rate of firings of stimulated neurons; the same equation, however, can be conceivably applied to a vast variety of phenomena. The procedures followed to solve the problems that arise in connection with this equation could be extended to more general types of non linear equations.The research reported in this document has been sponsored in part by the Air Force Office of Scientific Research under Contract no. AF EOAR 65-44 through the European Office of Aerospace Research, OAR, United States Air Force.     

Neuronal localization of dopamine,noradrenaline, and 5-hydroxytryptamine in the central and peripheral nervous system ofLumbricus terrestris (L.)

Summary Fluorescence microscopical studies with the procedure of Falck and Hillarp have confirmed previous observations concerning the appearance of neurones with green and yellow specific fluorescence in the central and peripheral nervous system ofLumbricus terrestris.Chemical estimates show that the fluorescent neurones contain the primary catecholamines dopamine and noradrenaline, in addition to an indolamine, presumably 5-hydroxytryptamine. Rude's opinion that dopamine is present in a concentration twice that of noradrenaline is confirmed.Microspectrofluorometric analyses of the neurones displaying green specific fluorescence show two types of neurones, one presumably containing dopamine (mainly the receptor cells, certain small and some of the large cells in the cerebral ganglion). Some of the large cells of the cerebral ganglion and the bipolar cells near the base of the second segmental nerve in the ventral nerve cord show characteristics compatible with the simultaneous presence of both noradrenaline and dopamine in them.This work was supported by grants from the Helge Ax:son Johnson Foundation and was carried out within a reasearch organization sponsored by the Swedish Medical Research Council (projects No. B71-14X-2321-04A, B71-14X-712-06A, and B71-14X-56-07A).     

Binding of catecholamines and related compounds to proteins after formaldehyde gas treatment

Summary Primary catecholamines e.g. dopamine and noradrenaline easily condense with formaldehyde in both tissues and model protein layers, yielding strongly fluorescent 3,4-dihydroisoquinolines. This reaction can be used for the histochemical demonstration of biogenic monoamines according to the method of Falck and Hillarp. The investigation reported presents data which suggest that the isoquinolines formed are mainly enclosed in or adsorbed on the network formed by the formaldehyde tanning of the protein. The products seem not to be chemically bound. No difference was found in the binding of the different biogenic catecholamines. The degree of binding can be reinforced by prolonged formaldehyde gas treatment and the use of more humid gas, which might prove useful when this binding method is employed for the autoradiography of catecholamines and 5-hydroxytryptamine. The possibilities of chemical binding are discussed.     

Spectral absorption by screening pigment granules in the compound eyes of a moth and a wasp

Summary The spectral absorption by single ommin containing pigment granules or clusters of granules from compound eyes was measured spectrophotometrically between 300 and 700 nm. The measurements were made on fresh and fixed slices from compound eyes of Celerio euphorbiae and Vespa spec. In the visible part of the spectrum there is an absorption maximum between 540 and 550 nm, situated nearly 30 nm more towards the red than that of pure ommin in solution. A frequently found side maximum of variable height at about 450 nm is probably caused by oxidized xanthommatin occurring additionally within the granules. The absorption increases from 350 nm towards shorter wavelengths, and gradually declines between 550 and 700 nm.This work was supported partly by the Swedish Medical Research Council, Stiftelsen Gustaf och Tyra Svenssons Minne, Reservationsanslaget, and partly by the Air Force Office of Scientific Research through the European Office of Aerospace Research, OAR, United States Air Force under grant number EOOAR-68-0036.