Ribbon synapses of the retina

Vision is a highly complex task that involves several steps of parallel information processing in various areas of the central nervous system. Complex processing of visual signals occurs as early as at the retina, the first stage in the visual system. Various aspects of visual information are transmitted in parallel from the photoreceptors (the input neurons of the retina) through their interconnecting bipolar cells to the ganglion cells (the output neurons). Photoreceptors and bipolar cells transfer information via the release of the neurotransmitter glutamate at a specialized synapse, the ribbon synapse. Although known from early days of electron microscopy, the precise functioning of ribbon synapses has yet to be explained. In this review, we highlight recent advances towards understanding the molecular composition and function of this enigmatic synapse.This study was supported by a grant from the Deutsche Forschungsgemeinschaft (BR 1643/4-1) to J.H.B.     

Evidence for ecological adaptation of circadian systems

Summary Comparative studies on the circadian activity rhythm of five neotropical bat species show, that even exclusively tropical Chiroptera yield considerable differences with respect to their light-induced variation of the period length and the time needed for re-entrainment after phase shifts of the Zeitgeber-LD (Figs. 2 and 3). Species with a wide variation range of the period length resynchronize very fast and vice versa. Both reactions suggest the circadian system to have a higher plasticity or a higher sensibility for the exogenous components respectively. The sensibility for these exogenous components is, according to the hitherto results, in general higher within insectivorous Chiroptera species than within frugivorous/nectarivorous to omnivorous species. It is discussed, whether this can be interpreted as a specific adaptation to the demands of a predatory way of life.Dedicated to Prof. Dr. F.P. Möhres on the occasion of his 65th birthdayThis work was supported by the Deutsche Forschungsgemeinschaft (Er 59/1-5)     

Immunohistochemical evidence for the presence of peptides derived from proenkephalin,prodynorphin and proopiomelanocortin in the guinea pig pineal gland

Summary By using a plethora of region-specific antisera, this light microscopic immunohistochemical study revealed that derivatives from the three opioid precursors, i.e. proenkephalin, prodynorphin and proopiomelanocortin are differentially distributed in the pineal gland of guinea pig. Various molecular forms of immunoreactive opioid peptides derived from proenkephalin or prodynorphin were prosent in a minority of pinealocytes as well as in nerves. In contrast to this dual distribution pattern of opioid-active peptides, the opioid-inactive derivative from proopiomelanocortin, alpha-melanocyte stimulating hormone, was exclusively present in a large proportion of pinealocytes. A multiple and differential origin and function of opioidergic pineal innervation involving sympathetic, parasympathetic and sensory components is suggested. -MSH is proposed as a pineal hormone which may act in concert with melatonin to regulate pineal rhythms or may function like MSH of pituitary origin.Dedicated to Professor Dr. T.H. Schiebler on the occasion of his 65th birthdaySupported by the Deutsche Forschungsgemeinschaft (grant Schr 283/2-1 and We 910/2-1) and by Stifterverband für die Deutsche Wissenschaft, Schering AG and Freunde der Universität Mainz     

Response of the cercus-to-giant interneuron system in crickets to species-specific song

Summary In addition to their high frequency stridulatory sound, crickets (Gryllus campestris) also produce low frequency airborne vibrations resulting from the strokes of the crickets' wings closing and opening during stridulation. Giant interneurons of the ventral cord, which receive inputs from cereal hairs, respond to these low frequencly components of cricket song up to a distance of some ten cm. The discharges are correlated to the time course of the acoustic stimulation, therefore allowing the transmission of the time patterns (syllables, chirps) of calling song, rivalry song and even courtship song. With simultaneous recording from both sides of the abdominal nerve cord, synchronous or alternating discharges can be detected, correlated either with syllables or intervals. The response mode depends on the position of the two individuals in relation to each other. Recording directly from singing males demonstrates interneuron response to selfgenerated signals, too. A possible communicative function of the system is discussed.Supported by the Deutsche Forschungsgemeinschaft as part of the program Neurale Mechanismen des Verhaltens (Da 61/8, 61/9)We thank Dr. F. Huber and Dr. H. Scharstein for helpful comments.     

The multifunctional sensory complex in the antennae ofAllacma fusca (Insecta)

Summary The antennal-tip sensory complex inAllacma fusca (Collembola) was reconstructed from serial ultrathin sections. The complex contains 16 sensory cells which belong to three spatially separated subunits: (1) a sensory hair; (2) a cuticular protrusion, containing two highly-specialized dendritic outer segments from a single sensory cell encapsulated by an enveloping cell and a gland cell (Figs. 4 and 5); and (3) two sensory cells with partly lamellated, double dendritic outer segments located well away from the cuticular surface (Fig. 3). A phylogenetic evaluation reveals a mosaic of apparently primitive characters, and trends toward higher structural complexity as well as toward reduction (Table 1). There is support for a hypothesis that the receptors of the sensory hair are chemosensitive. The functional interpretation of the other components of the complex is problematic due to their unusual structural properties. The sensory cell beneath the cuticular protrusion is considered to be proprioceptive.Supported by the Deutsche Forschungsgemeinschaft (Al 56/6)     

Laryngeal nerve activity during pulse emission in the CF-FM bat,Rhinolophus ferrumequinum

Summary The activity of the external (motor) branch of the superior laryngeal nerve (SLN), innervating the cricothyroid muscle, was recorded in the greater horseshoe bat,Rhinolophus ferrumequinum. The bats were induced to change the frequency of the constant frequency (CF) component of their echolocation signals by presenting artificial signals for which they Doppler shift compensated. The data show that the SLN discharge rate and the frequency of the emitted CF are correlated in a linear manner.Abbreviations SLN Superior laryngeal nerve - RLN Recurrent laryngeal nerve - DCS Doppler compensation system - CF Constant frequency - FM Frequency modulation Supported by grants of the Deutsche Forschungsgemeinschaft (DFG), Az.: Schu 390/1, /2 and SFB 45We are indebted to Dipl.-Ing. H. Zöller for providing the computer programs. We want to thank H. Hahn and A. Polotzek for technical help.     

Perceptual spatial frequency — Orientation surface: Psychophysics and line element theory

We have investigated the discriminability of gratings which simultaneously vary in spatial frequency and orientation. Thirteen and nine reference gratings were used with two observers, and bivariate discrimination probability surfaces were determined around each grating. These data were then fitted to a general bivariate Gaussian function. The results clearly demonstrate local separability in this log frequency and orientation discrimination domain. Our results also show that the factor contributing most to the non-Euclidean nature of such frequency/orientation discrimination is orientation anisotropia, although we also find some evidence for smaller changes in the associated Riemannian line-element at different frequency ranges. These results cast doubt upon claims for a pseudo line-element for frequency discrimination based upon the nonlinear outputs of a fixed set of detectors.Study supported by Fraunhofer Gesellschaft Grant INSAN I 0784-V-6385 and Guest Professorship Mu 93/103-1 for TC by Deutsche Forschungsgemeinschaft     

Improvement of the method of Karnovsky and Roots for the histochemical demonstration of acetylcholinesterase

Summary The aqueous reaction medium of Karnovsky and Roots for the demonstration of acetylcholinesterase (AChE) activity was modified to obtain, a gel-incubation medium that prevents diffusion of the reaction product and the soluble part of AChE into the incubation medium and along the plane of the section. The incubation medium contained 27% polyvinyl alcohol and suitable concentrations of the components of the medium of Karnovsky and Roots. The application of this incubation medium to sections of rat hippocampus resulted in intense staining, with the reaction products being mainly localized in fibrous structures.Supported by the Deutsche Forschungsgemeinschaft (Ku 541/2-1)     

Simultaneous tracking of flagellates in real time by image analysis

The hardware and software components are described to track many flagellates simultaneously in real time. The technique is based on on-line digitization of four frames taken at 80 ms intervals and stored in a specialized video memory. The outline and position of each organism are determined by chain coding and followed through the recorded series of images. The movement vectors of all organisms tracked are stored in the form of deviation angles from a predefined stimulus (light or gravity) direction and the distance each organism has moved in the time interval determined by the hardware clock of the computer. Subsequent programs allow one to determine circular histograms of movement directions and velocities in dependence of the movement direction. Examples of several orientation patterns are given for both photoorientation and gravitaxis in the dinoflagellate, Peridinium gatunense.Supported by grants from the Deutsche Forschungsgemeinschaft (SFB 305, Marburg and Ha 985/6-1, Erlangen)     

Innervation of the substantia nigra

This review describes inputs to neurons in the substantia nigra and contrasts them with the action of agonists for the putative receptors through which they act. Special emphasis is placed on gamma-aminobutyric acid (GABA) afferents. Dopamine released from the somato-dendritic compartment of dopamine neurons and endocannabinoids released from dopamine and GABA neurons serve as retrograde signals to modulate GABA release. The release may be fostered by Ca²⁺ release from intracellular Ca²⁺ stores, which in turn may be influenced by the inputs.The studies summarized in this review were supported by the Deutsche Forschungsgemeinschaft (FOR 302/TP-B1)     

Peripheral auditory tuning for fine frequency analysis by the CF-FM bat,Rhinolophus ferrumequinum

Summary For echolocation,Rhinolophus ferrumequinum emits orientation sounds, each of which consists of a long constant-frequency (CF) component and short frequency-modulated (FM) components. The CF component is about 83 kHz and is used for Doppler-shift compensation. In this bat, single auditory nerve fibers and cochlear nuclear neurons tuned at about 83 kHz show low threshold and very sharp filter characteristics. The slopes of their tuning curves ranged between 1,000 and 3,500 dB/octave and their Q-10 dB values were between 20 and 400, 140 on the average (Figs. 3–5). The peripheral auditory system is apparently specialized for the reception and fine frequency analysis of the CF component in orientation sounds and Doppler-shift compensated echoes. This specialization is not due to suppression or inhibition comparable to lateral inhibition, but due to the mechanical specialization of the cochlea. Peripheral auditory neurons with the best frequency between 77 and 87 kHz showed not only on-responses, but also off-responses to tonal stimuli (Figs. 1, 2, and 6). The off-responses with a latency comparable to that of N₁-off were not due to a rebound from either suppression or inhibition, but probably due to a mechanical transient occurring in the cochlea at the cessation of a tone burst.We thank Alexander von Humboldt Stiftung, Deutsche Forschungsgemeinschaft (Grant No. Ne146/6-8), Stiftung Volkswagenwerk (Grant No. 111858), and American National Science Foundation (Grant No. 40018 and BMS 75-17077) for their support for our cooperative work.     

Differential tissue distribution of the Invs gene product inversin

Nephronophthisis is a common genetic cause of end-stage renal disease in childhood. Recently, Invs was identified as the gene mutated in the infantile form of nephronophthisis. Humans with nephronophthisis develop a large number of extrarenal manifestations, including situs variations, anomalies of the hepatobiliary system, retinal degeneration and cerebellar ataxia. Mice homozygous for a mutation in the Invs gene (inv mouse) die during the first week after birth as a result of renal and liver failure. Although organ anomalies have been characterized in human nephronophthisis and the inv mouse, little is known about the tissue expression of the Invs gene product, inversin. We have used laser confocal microscopy of paraffin-embedded murine tissue sections to provide the first detailed characterization of the distribution of inversin in various organs. Our results show that inversin is localized to distal tubules in the kidney, hepatic bile ducts, acinar and ductal pancreatic cells, epithelial intestinal cells, splenic germinal centres, bronchiolar epithelial cells, dendrites of cerebellar Purkinje cells, retinal neural cells and spermatocytes and spermatids in the testis. The localization of inversin in distal tubules in the kidney and in extrarenal tissues suggests that the expression of this protein has an important function in a variety of organs. Further studies are required to understand the way in which mutations in the Invs gene lead to the multi-organ pathology of inv mouse and human nephronophthisis. J.N. acknowledges funding from the Deutsche Forschungsgemeinschaft (Nu 118/1-1, Nu 118/3-1) and the intramural research program of the University Hospital of Essen (IFORES). A. K. acknowledges funding from the Deutsche Forschungsgemeinschaft (Kr 1108/2-2).     

Interdependent development of blood vessels and organs

The cardiovascular system is the first functional organ in the vertebrate embryo, and many organs start to develop adjacent to cells of the cardiovascular system. Endothelial cells (EC) form the inner cell lining of blood vessels and represent the major cell type that interacts with developing organs. On the one hand, EC provide organs with signals. These signals determine the location, differentiation and morphology of an organ. On the other hand, EC receive signals from the organ-specific cell types. Such signals give EC organ-specific features that the organ needs to interact with the circulatory system. This review provides the reader with specific examples of an interdependent development of organs and blood vessels.Eckhard Lammert and Ganka Nikolova were supported by the Deutsche Forschungsgemeinschaft DFG (La1216/2–1)     

Cytochemical demonstration of aspartate aminotransferase in the mossy-fibre system of the rat hippocampus

Summary A lead-salt procedure was used for the cytochemical demonstration of aspartate aminotransferase (AT) in the CA3 of the rat hippocampus. Cytoplasmic- and mitochondrial-AT isoenzymes were demonstrated in different neuronal structures, but not in astrocytes. Of special interest was the localization of cytoplasmic AT in most mossy-fibre boutons. This might indicate that cytoplasmic AT is responsible for the production of releasable glutamate in the glutamatergic/aspartatergic mossy fibre system.Supported by the Deutsche Forschungsgemeinschaft (Ku 541/2-1)     

Glial cells in the pineal gland of mice and rats

Summary Antigenic markers characteristic of astrocytes and their differentiative states (i.e., glial fibrillary acidic protein (GFAP), vimentin, and M1 and C1 antigens) were investigated in the pineal gland of mouse and rat using double immunolabeling techniques. In both species the socalled interstitial cells as characterized by TEM were shown to be astrocytes, since they expressed vimentin, but neither fibronectin (a marker for fibroblasts and endothelial cells) nor the neuron-specific L1 antigen or tetanus toxin receptors. Subpopulations of vimentin-positive pineal astrocytes were also GFAP- and C1- antigen-positive. M1- antigenpositive cells were not detected.It is concluded that a considerable proportion of interstitial cells in the pineal gland of rat and mouse are immature astrocytes which, in contrast to other parts of the central nervous system, persist into adulthood.Supported in part by Deutsche Forschungsgemeinschaft (Scha 185/9-4)S.-K. Huang was a recipient of a Humboldt Foundation fellowship.     

Neural connections between the brain and the pineal gland of the golden hamster (Mesocricetus auratus)

Summary Neurons projecting from the brain to the pineal gland via the pineal stalk were investigated in the golden hamster with the use of the retrograde horseradish-peroxidase tracing method both in vivo and in vitro. Labelled perikarya were observed in the medial and lateral habenular nuclei as well as in the posterior commissure. Single cells located in the ependymal lining of the pineal- and suprapineal recesses were also retrogradely labelled. These results show that a distinct central innervation of the pineal gland exists in the golden hamster, in agreement with findings in other mammalian species investigated by means of a similar methodology. In addition, also direct signals from the cerebrospinal fluid to the parenchyma might be conducted via cells located within the ependymal layer of the pineal- and suprapineal recesses.This study was supported by grants from the Deutscher Akademischer Austauschdienst to M.M. (312/dk-4-is), the Deutsche Forschungsgemeinschaft to H.W.K. (Ko 758/2-2, 2-3), and the Carlsberg Foundation     

CSF-contacting and other somatostatin-immunoreactive neurons in the brains of Anguilla anguilla,Phoxinus phoxinus,and Salmo gairdneri (Teleostei)

Summary A system of somatostatin-immunoreactive neurons was demonstrated in the brains of the eel, Anguilla anguilla, the European minnow, Phoxinus phoxinus, and the rainbow trout, Salmo gairdneri, by means of the light-microscopic indirect immunoperoxidase technique.In the anterior periventricular nucleus, somatostatin-immunoreactive cerebrospinal fluid (CSF)-contacting neurons display intensely stained intraventricular dendritic protrusions, perikarya, and axonal processes. The latter taper into a somatostatin-immunoreactive fiber plexus extending to the infundibulum, the proximal neurohypophysis, and the lateral and mammillary recesses.In addition, somatostatin-immunoreactive neurons were demonstrated in the magnocellular preoptic, entopeduncular and dorsolateral thalamic nuclei, further in the pretectal area and the ventrolateral tegmentum. Somatostatin-immunoreactive fiber bundles project via the stria medullaris toward the habenular nucleus; they also course in the dorsomedial-ventrolateral direction at the level of the pretectal-tegmental area, and within the ventral and dorsal tegmentum.The presence of somatostatin in a variety of different neurons of the teleost brain is discussed in connection with their tentative inhibitory function. The CSF-contacting neurons of the anterior periventricular nucleus are supposed to function as sensors that pass information from the CSF to the somatostatin system of the hypothalamus and/or other components of the neuroendocrine apparatus.Dedicated to Professor Helmut Leonhardt, Kiel, on the occasion of his 65^th birthdayThis investigation was supported by grants from the Deutsche Forschungsgemeinschaft to A.O. (Ok 1/25-3) and H.W.K. (Ko 758/2-2)On leave of absence from the second Department of Anatomy, Semmelweis OTE, Budapest, Hungary     

Genetic considerations on association between HLA and disease

Summary Blood specimens from a random sample of 981 South African Negroid females were typed electrophoretically inter alia for their G-6-PD phenotypes. The allele frequency for Gd^B and Gd^nonB was found to be 0.8126 and 0.1874 respectively. Calculating the number of individuals expected for each phenotypic class, a highly significant deviation from the Hardy-Weinberg equilibrium became manifest, i.e. there was a deficit of 24.6% of heterozygotes and an excess of 12.3% of each of the two classes of homozygotes.Several possible reasons for this discrepancy e.g. the effects of pooling sub-samples, selection and misclassifications due to insufficient staining were examined and were found not to be likely explanations for the observed phenomenon. Instead, the result is interpreted as due to only 3–4 stem cells which give rise to the haematopoetic system in man.Part of this work was presented at the biennial Congress of the Deutsche Gesellschaft für Anthropologie und Humangenetik held in Vienna, September 22–25, 1975Supported by a research fellowship (1975/76) granted by the Alexander von Humboldt-Stiftung, Bonn-Bad GodesbergSupported by the Deutsche Forschungsgemeinschaft (DFG), Bonn-Bad Godesberg     

DNA fingerprinting with oligonucleotides can differentiate cell lines derived from the same tumor

Summary Oligonucleotide fingerprinting was applied to investigate the relatedness of several cell lines that were established between 1973 and 1977 from a teratocarcinoma. We were able to distinguish cell lines derived at different times. In addition, sublines from one cell line (PYS-2) could be discriminated by using a combination of different probes. Therefore multilocus fingerprinting with oligonucleotides is a useful method for monitoring changes in cell lines kept in culture for many generations. This work was supported by the Deutsche Forschungsgemeinschaft (OB 66/2-1) and by the VW-Stiftung.     

Spatial organization of transport domains and subdomain formation in the plasma membrane of Chara corallina

Pattern formation mechanisms in developing organisms determine cellular differentiation and function. However, the components that interact during the manifestation of a spatial pattern are in general unknown. Characean algae represent a model system to study pattern formation. These algae develop alternating acid and alkaline transport domains that influence the pattern of growth. In the present study, it will be demonstrated that a diffusion mechanism is implicated in acid and alkaline domain formation and this growth pattern. Experiments on the characean growth pattern were performed that resulted in pronounced, however, unpredictable modifications in the original pattern. A major component involved in this pattern-forming mechanism emerged from the nonlinear kinetics of the H⁺-ATPase that is located in the plasma membrane of these algae. Based on these kinetics, a mathematical model was developed and numerically analyzed. As a result, the contribution of a diffusional component to the characean acid/alkaline pattern appeared most likely.This work was supported by the Deutsche Forschungsgemeinschaft (grant #571 1/1) to JF.