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I Kupfermann

Publications and source records attributed to I Kupfermann.

At least 91 records · Page 5Linked to original sources

The small cardioactive peptides A and B of Aplysia are derived from a common precursor molecule.

We have identified cells in the central nervous system of the marine mollusc Aplysia that react with antibody raised against the small cardioactive peptide B (SCPB). Antisera to this neuropeptide stained a subset of central neurons that include the large identified buccal neurons, B1 and B2. The distribution of SCP-containing neurons was used in a strategy to isolate a cDNA clone encoding the precursor protein for the peptide. RNA from neurons B1 and B2 and from cells that did not stain with SCPB antisera was used to direct the synthesis of radiolabeled cDNA probes. A cDNA clone complimentary to mRNA specifically expressed in the B1 and B2 cells was isolated by differentially screening a buccal cDNA library with these probes. The cloned cDNA segment is 1394 nucleotides in length and contains a 408-base-pair open reading frame. The predicted precursor protein is composed of 136 amino acids and has a characteristic hydrophobic leader sequence. The sizes of the precursor protein with and without this leader sequence agree with in vivo and in vitro labeling studies. The amino acid sequences for SCPB and a related peptide, SCPA, are present and are flanked by known proteolytic processing sites.

Amino Acid Sequence↗

Biochemical and immunocytological localization of molluscan small cardioactive peptides in the nervous system of Aplysia californica.

High pressure liquid chromatography (HPLC) followed by bioassay on isolated snail hearts were used to locate two related peptides, termed small cardioactive peptides A and B (SCPA and SCPB) in each of the central ganglia of Aplysia. The peptides are most concentrated in the buccal ganglia, the ganglia involved in the control of feeding movements. Immunocytology with antisera raised to conjugated SCPB stained three groups of neurons in the buccal ganglia. One group consisted of relatively small neurons that were tightly clustered. The second group was comprised of larger neurons that were more scattered. The third group was made up of several neurons including the two largest in the ganglia, identified cells B1 and B2. B1 and B2 and other neurons in this group innervate the gut by way of the esophageal nerve. HPLC-bioassay of single, individually dissected B1 or B2 neurons demonstrated that the two peptides are present in a single cell. For B2, but not B1, choline injected into the cell body was converted to the conventional transmitter, acetylcholine. This indicates that, in addition to the two peptides, B2 also contains choline acetyltransferase, and raises the possibility that acetylcholine and the SCPs may act as co-transmitters in B2. Strong immunocytological staining of fibers and varicosities was observed in the neuropilar region of the cerebral, pleural, pedal, and abdominal ganglia. In addition to the buccal ganglia, immunoreactive neurons were observed in all of the other central ganglia. The high concentration of the SCPs and the relatively large number of immunoreactive neurons in the buccal ganglion suggest a particularly important role of these peptides specifically in feeding behavior. However, the widespread occurrence of the SCPs in fibers and neuronal cell bodies throughout the nervous system suggests that these peptides also may have additional behavioral functions in Aplysia.

Aplysia↗

Internal stimuli enhance feeding behavior in the mollusc Aplysia.

The hypothesis that subsatiating levels of internal food stimuli can arouse and potentiate feeding behavior was examined in the mollusc Aplysia californica. Animals were fed a small quantity of seaweed and their latencies to show biting responses were determined after food arousal was permitted to partially decay. Control animals were stimulated with food, but were not permitted to ingest it, or were fed nonnutritive glass-fiber filter paper. Compared to controls, animals that were fed showed significantly shorter latencies to respond when tested up to 80 min after previous exposure to food. These results indicate that internal stimuli can function like external stimuli to enhance responsiveness to food and suggest the hypothesis that satiation may be viewed as an interactive process involving the interplay of excitatory and inhibitory influences arising from the alimentary system.

Animals↗

Evidence for parallel actions of a molluscan neuropeptide and serotonin in mediating arousal in Aplysia.

The neuropeptide designated SCPB (small cardioactive peptide B), the sequence of which has recently been determined, was found in the accessory radula closer muscle, a muscle involved in biting movements. The ganglia and nerves that innervate the accessory radula closer muscle also contain SCPB. At nanomolar concentrations, it enhances the contractions of the muscle. The effect of SCPB on the muscle resembles the effect of an identified serotonergic neuron that previously was shown to mediate behavioral effects that reflect a food arousal state in Aplysia. Like serotonin, SCPB enhances contractions by a postsynaptic action, which appears to involve an increase in cAMP levels in the muscle. Our findings suggest that parallel peptidergic and serotonergic pathways may mediate similar aspects of arousal in Aplysia.

Animals↗

Lesion of a serotonergic modulatory neuron in Aplysia produces a specific defect in feeding behavior.

The serotonergic metacerebral cells (MCCs) of Aplysia were destroyed by intracellular injection of proteolytic enzyme. MCC-lesioned animals showed alterations of biting responses compared to MCC-sham and B-cell-lesioned control animals, as well as to their own preoperative behavior. The alterations of biting responses included a prolongation of the duration of radula protraction and a lengthening of interbite interval. No changes were observed in non-biting feeding responses and in behaviors unrelated to feeding.

Animals↗

Activity of an identified serotonergic neuron in free moving Aplysia correlates with behavioral arousal.

Extracellular recordings of the metacerebral cell (MCC), a serotonergic neuron in Aplysia, were obtained in free moving, undrugged animals. MCC activity was evoked by exposure to food. Arousal level was manipulated by satiating the animals or exposing them to a noxious stimulus. We found that the amount of evoked MCC activity correlated with the level of arousal of the animal.

Animals↗

Ultrastructure of a histaminergic synapses in Aplysia.

The ultrastructure of histaminergic synaptic terminals was studied by the means of intrasomatic injection of horseradish peroxidase into the identified histaminergic neuron C2 of Aplysia. The axonal tree of C2 was found to consist, in part, of varicosities that display putative release sites similar in morphology to those described in other neurons in Aplysia. The varicosities contain at least two populations of vesicles: a conspicuous class of of large vesicles with an electron-dense core that almost fills the entire vesicle and a heterogeneous class of large and small electron-lucent vesicles. The small lucent vesicles preferentially cluster near active zones.

Animals↗

Modulatory actions of neurotransmitters.

A relatively shorttime ago the individual neuron was viewed as functioning by means of the simple summation of brief inhibitory or excitatory events. The complexity of the nervous system was the outcome largely of the connections between neurons. Recent studies have uncovered a new set of phenomena that indicate that relatively complex information processing may occur at the level of the individual neuron. For example, rather than producing additive effects, synaptic inputs can produce multiplicative effects that serve to alter the gain of the system. In addition, synaptic inputs may be able to alter specific classes of other inputs selectively. One could envision that each class of synaptic input to a cell could be selectively depressed or enhanced by a corresponding modulatory input. Since the modulatory actions can be transmitted intracellularly via second messengers, an extensive array of presynaptic connections may be unnecessary. It remains to be determined which of the modulatory phenomena currently reviewed are functionally important and which are only pharmacological or experimental curiosities. Are there any common attributes of the various synaptic phenomena that have been termed modulatory? The most common features of modulatory synaptic effects are long-duration of action and contingent action. Contingent action refers to the property that modulatory transmitters often have little or no effects in themselves, but instead they alter the effects of other events. Long-duration and contingent action endow modulatory effects with properties ideally suited to the control of behavioral modulations such as learning, motivational state, arousal, and sensitization. While there is no necessary connection between behavioral modulation and neural modulation, the available evidence from invertebrates suggests that there often is a connection. It is likely that examples of neuromodulation in vertebrates also will be tied to behavioral modulation.

Animals↗

Response properties and synaptic connections of mechanoafferent neurons in cerebral ganglion of Aplysia.

1. The cells of two clusters of small neurons on the ventrocaudal surface of each hemicerebral ganglion of Aplysia were found to exhibit action potentials following tactile stimuli applied to the skin of the head. These neurons appear to be mechanosensory afferents since they possess axons in the nerves innervating the skin and tactile stimulation evokes spikes with no prepotentials, even when the cell bodies are sufficiently hyperpolarized to block some spikes. The mechanosensory afferents may be primary afferents since the sensory response persists after chemical synaptic transmission is blocked by bathing the ganglion and peripheral structures in seawater with a high-Mg2+ and low-Ca2+ content. 2. The mechanosensory afferents are normally silent and are insensitive to photic, thermal, and chemical stimuli. A punctate tactile stimulus applied to a circumscribed region of skin can evoke a burst of spikes. If the stimulus is maintained at a constant forces, the mechanosensory response slowly adapts over a period of seconds. Repeated brief stimuli have little or no effect on spike frequency within a burst. 3. Approximately 81% of the mechanoafferent neurons have a single ipsilateral receptive field. The fields are located on the lips, the anterior tentacles, the dorsal portion of the head, the neck, or the perioral zone. Because many cells have collateral axons in the cerebral connectives, receptive fields elsewhere on the body are a possibility. The highest receptive-field density was associated with the lips. Within each area, receptive fields vary in size and shape. Adjacent fields overlap and larger fields frequently encompass several smaller ones. The features of some fields appear invariant from one animal to the next. A loose form of topographic organization of the mechanoafferent cells was observed. For example, cells located in the medial cluster have lip receptive fields, and most cells in the posterolateral portion of the lateral clusters have tentacle receptive fields. 4. Intracellular stimulation of individual mechanoafferents evokes short and constant-latency EPSPs in putative motor neurons comprising the identified B-cell clusters of the cerebral ganglion. On the basis of several criteria, these EPSPs appear to be several criteria, these EPSPs appear to be chemically mediated and are monosynaptic. 5. Repetitive intracellular stimulation of individual mechanoafferent neurons at low rates results in a gradual decrement in the amplitude of the EPSPs evoked in B cluster neurons. EPSP amplitude can be restored following brief periods of rest, but subsequent stimulation leads to further diminution of the response. 6. A decremented response cannot be restored by strong mechanical stimulation outside the receptive field of the mechanoafferent or by electrical stimulation of the cerebral nerves or connectives...

Animals↗

Functional role of serotonergic neuromodulation in Aplysia.

The serotonergic metacerebral cell (MCC) of the mollusk Aplysia produces slow synaptic potentials in motor neurons of the buccal muscle, and increases the rate of ongoing rhythmic burst output of the buccal ganglion. In addition, the MCC acts peripherally to enhance the strength of buccal muscle contractions that are produced by firing of motor neurons. The potentiation of contraction is not associated with any detectable changes of resting membrane potential of muscle cells. Although MCC activity produces a small enhancement of excitatory junctional potentials, several experiments clearly indicate that the MCC has a direct potentiating effect on excitation-contraction coupling. The data suggest that potentiation of contraction might be mediated by cAMP. For example, activity of the MCC enchances the rate of accumulation of cAMP in buccal muscle, application of phosphodiesterase resistant analogs of cAMP potentiates muscle contraction, and a phosphodiesterase inhibitor enhances the effect of MCC stimulation. Recordings from free-moving animals indicate that the MCC becomes activated by exposure of the animal to food stimuli, and that the activation parallels the presence of a food-arousal state. Food-arousal is characterized by enhanced strength and increased frequency of biting responses. Both these effects can result from activity of the MCC. Thus, in this system, modulatory synaptic actions function to provide the substrate for a type behavioral modulation.

Animals↗