Search PubMed⌕ Search

Biomedical subjects

M Eckert

Publications and source records attributed to M Eckert.

At least 37 records · Page 2Linked to original sources
PubMed1999

Quantification of periviscerokinin-1 in the nervous system of the American cockroach, Periplaneta americana. An insect neuropeptide with unusual distribution.

This study was undertaken to reveal the quantitative distribution of the insect neuropeptide periviscerokinin-1 (Pea-PVK-1) in the central nervous system of Periplaneta americana and to demonstrate that neurons stained in a previous immunohistochemical study contain authentic Pea-PVK-1. For this, we combined ELISA, HPLC, and MALDI-TOF mass spectrometry. The high specificity of the used antiserum enabled the quantification of Pea-PVK-1 in unseparated tissue extracts. No cross-reactivities with other insect neuropeptides were detected in ELISA. Only two immunoreactive fractions, coeluting with synthetic Pea-PVK-1 in its oxidized and nonoxidized form, were found in HPLC-separated extracts of the brain, suboesophageal ganglion, metathoracic ganglion, second abdominal ganglion with or without perisympathetic organ, and terminal ganglion. By using MALDI-TOF mass spectrometry, we were able to confirm the existence of authentic Pea-PVK-1 in these fractions. The abdominal perisympathetic organs contained 6.3 pmol Pea-PVK-1 per animal; another 1.3 pmol were found in the abdominal ganglia. More than 90% of the total 8.2 pmol in the central nervous system was found in the abdominal ganglia and their perisympathetic organs. The corpora cardiaca and corpora allata did not contain immunoreactive material, suggesting that Pea-PVK-1 is not released by the cephalic neurohaemal system. The quantitative distribution of Pea-PVK-1 differs considerably from that of other known insect neuropeptides.

Animals↗
PubMed1999

Periviscerokinin-like immunoreactivity in the nervous system of the American cockroach.

A highly specific polyclonal antiserum has been raised against periviscerokinin, the first neuropeptide isolated from the perisympathetic organs of insects (Predel et al. 1995). In this study, two different neuronal systems with periviscerokinin-like immunoreactivity were distinguished in the central nervous system of the American cockroach: (1) An intrinsic neuronal network, restricted to the head-thoracic region, was formed by intersegmental projecting neurons of the brain, suboesophageal ganglion and metathoracic ganglion. In addition, groups of local interneurons occurred in the proto- and tritocerebrum. (2) A typical neurohormonal system was stained exclusively in the abdomen; it was represented by abdominal perisympathetic organs which were supplied by three cell clusters located in each unfused abdominal ganglion. As revealed by nickel backfills, most neurons with axons entering the perisympathetic organs contained a periviscerokinin-like peptide. Immunoreactive fibres left the perisympathetic organs peripherally, innervated the hyperneural muscle and ran via the link nerves/segmental nerves to the heart and segmental vessels. All visceral muscles innervated by periviscerokinin-immunoreactive fibres were shown to be sensitive to periviscerokinin, whereas the hindgut gave no specific response to this peptide.

Animals↗
PubMed1999

The unique neuropeptide pattern in abdominal perisympathetic organs of insects.

We successfully isolated and identified the abundant neuropeptides of the abdominal perisympathetic organs of the American cockroach, including all myoactive compounds. Peptide sequence analysis and mass spectrometry of abundant substances that were not bioactive in different muscle assays yielded the following sequences: TDPLWQLPGAHLEQYLS-NH2 (Pea-YLS-amide), AFLTLTPGSHVDSYVEA-OH (Pea-VEAacid), and SDLTWTYQSPGDPTNSKN-OH (Pea-SKNacid). The given structures led to the conclusion of an unique neuropeptide pattern in abdominal perisympathetic organs. We confirmed this assumption with immunocytochemical studies, using antisera raised against different myotropic neuropeptides of the abdominal perisympathetic organs. Moreover, mass spectrometric methods, developed for the investigation of single neurohemal organs, confirmed the neuropeptide pattern in these organs.

Abdomen↗
PubMed1998

Isolation of periviscerokinin-2 from the abdominal perisympathetic organs of the American cockroach, Periplaneta americana.

Using the isolated hyperneural muscle as bioassay, a novel myotropin was isolated from the abdominal perisympathetic organs of Periplaneta americana. This is the second neuropeptide identified from insect perisympathetic organs. Peptide sequence analysis and mass spectrometry yielded the following structure: Gly-Ser-Ser-Ser-Gly-Leu-Ile-Ser-Met-Pro-Arg-Val-NH2. This peptide, named periviscerokinin-2, was confirmed to be amidated by chemical synthesis, bioassay, and comparison of retention times between native and synthetic peptides. A highly specific antiserum was used to determine sites of synthesis in the abdominal ganglia. Besides periviscerokinin-1, periviscerokinin-2 is the only putative myotropic neurohormone from the abdominal perisympathetic organs that is effective in the nanomolar range. This confirms the hypothesis that the neurohormonal system of the ventral nerve cord is remarkably different from that of the brain.

Abdomen↗
PubMed1998

Species-specific action and distribution of tachykinin-related peptides in the foregut of the cockroaches Leucophaea maderae and Periplaneta americana.

Nine tachykinin-related peptides (TRPs) have been isolated from the brain and intestine of the cockroach Leucophaea maderae. In the present investigation, two of the nine TRPs, LemTRP 1 and 5, were tested for their ability to stimulate contractions in the foregut of the cockroaches L. maderae and Periplaneta americana in vitro. The two LemTRPs and the related locust peptide locustatachykinin I (LomTK I) induced contractions in the foregut of P. americana in a dose-dependent manner, but had no myostimulatory action in L. maderae. A half-maximal response for the LemTRPs and LomTK I was obtained at 5x10(-9)mol l-1. In both species, the neuropeptide proctolin stimulated foregut contractions. Using an antiserum to LomTK I, we demonstrated that in both species there are LomTK-like-immunoreactive (LomTK-LI) cell bodies and fibers within the ganglia and nerves of the stomatogastric nervous system. However, correlated with the species-specific action of the TRPs, we found efferent LomTK-LI nerve fibers supplying muscle fibers in the foregut of P. americana, but not in L. maderae. In both cockroach species, there is a rich supply of proctolin-immunoreactive fibers to the foregut muscle. Some of the LomTK-LI fibers supplying the P. americana foregut muscle contain co-localized proctolin immunoreactivity. These fibers appear to be derived from a large cell body in the frontal ganglion which also displayed co-localized immunoreactivities. Since TRP-containing neurons are restricted to the nerves and ganglia of the stomatogastric nervous system both in P. americana and L. maderae, TRPs may be involved in the control of foregut movements in both species, but in P. americana the control may be more complex with the additional peripherally projecting LomTK-LI neurons.

Animals↗
PubMed1997

Octopamine-containing neurons in the alimentary tract of the earthworm (Eisenia fetida).

Octopamine-containing nerve cells have been demonstrated in the enteric plexus of the earthworm (Eisenia fetida), applying immunocytochemistry and HPLC assay. A few octopamine-immunoreactive neurons occurred in the fore- and hindgut, whereas their number in the midgut was considerably greater. Octopamine levels detected by HPLC correlated with the distribution of octopamine-containing nerve cells. A regulatory role for these intrinsic octopaminergic neurons is suggested in the enteric plexus in the earthworm alimentary tract. This is the first report on the occurrence of octopamine-containing nerve cells in the peripheral nervous system of an invertebrate.

Animals↗
PubMed1997

[Hyperplastic polyps of the colon. An inhomogeneous group of pathologic-anatomic changes].

A total of 334 hyperplastic polyps of the colon of 266 patients were examined under epidemiologically, morphometrically and histochemically. Of these, 62 cases (23%) were associated with adenomas, 18 cases (7%) with carcinomas. Hyperplastic polyps are not entirely homogeneous, pathologically and -anatomically: Carcinoma-associated hyperplastic polyps are larger in diameter, the size and width of their crypts is greater, and they have a higher percentage of goblet cells with a higher amount of sialomucins as shown by HID/AB stain.

Adenomatous Polyps↗
PubMed1997

Review of the first 50 cases completed by the RACR mammography QA programme: phantom image quality, processor control and dose considerations.

The Mammography Quality Assurance Programme, recently established by the Royal Australasian College of Radiologists, has processed the first 50 applications. This programme, which closely follows the programme of the American College of Radiology (ACR), utilizes phantom film images, thermoluminescent dosimetry measurement of mean glandular dose, processor control charts, clinical images, equipment reports and required survey information to establish that a centre conforms to a minimum standard in mammography. The present paper describes the initial results of the first phantom images, dose measurements, processor control and survey information. A film review panel of six members has been trained in phantom film reading. Their evaluation of phantom films was compared with film readings by members of the ACR and was found to be in close agreement. Fifty films have been evaluated up to the present time with a failure rate of 26%. The major causes of failure were unacceptable film artefacts and poor contrast (as indicated by reduced fibre and mass visibility). A surprising result was the high failure in processing, where 23% of units reviewed had significant problems, including failure to keep the processor within required control limits. Only one centre recorded a mean glandular dose above 2 mGy with no centre over the 3 mGy limit. A review of the frequency of the quality control testing shows that the acceptance of quality assurance in mammography, while greater than in the initial stages of the ACR programme, is less than current US practice. These initial results for the accreditation process probably reflect an initial period of adjustment, as seen by the high pass rate achieved by centres that have resubmitted material to gain accreditation.

Accreditation↗
PubMed1996

A comparative immunocytochemical study using an antiserum against a synthetic analogue of the corpora cardiaca peptide Pea-CAH-I (MI, neurohormone D) of Periplaneta americana.

An antiserum against the octapeptide Pea-CAH-I, a member of the adipokinetic hormone/red pigment-concentrating hormone family, has been produced for immunocytochemical staining in insects and various other invertebrate species. The anti-Pea-CAH-I serum stains the glandular corpora cardiaca cells of those insect species that synthesize identical or structurally similar peptides. In the corpora cardiaca of species producing peptides with a different C-terminus, these cells remain unstained. Pea-CAH-I-like immunoreactivity has also been found in neurons of the central nervous system of all invertebrate orders studied. The antiserum recognizes the C-terminal sequence Pro-Asn-Trp-NH2 of the Pea-CAH-I molecule as established by enzyme immunoassay. The widespread Pea-CAH-I-like immunoreactivity in all nervous systems of the studied animals probably does not reflect the presence of Pea-CAH-I but the occurrence of peptides carrying similar epitopes.

Amino Acid Sequence↗
PubMed1996

Octopamine in the developing nervous system of the pond snail, Lymnaea stagnalis L.

Development of the octopaminergic system in the pond snail, Lymnaea stagnalis, was investigated by means of immunocytochemistry and radioenzymatic assay. The earliest octopamine-immunoreactive neurons appear at a late embryonic stage (E85) of development following metamorphosis. At this moment of development, the adult-like pattern of distribution and projection (arborization) characteristics of octopamine-immunoreactive neurone can already be observed. During hatching and postembryonic (juvenile) development the number of labelled neurons increases only within the ventro-medial cell groups of the cerebral ganglia, whereas the extent of varicose axon arborization of the labelled neurons increases gradually. No peripheral projections of the embryonic and postembryonic octopamine-immunoreactive neurons were observed. The postembryonic increase in number of immunoreactive neurons and development of axonal arborization is accompanied by a rapid, exponential enhancement of octopamine content of Lymnaea CNS, as detected radioenzymatically. A possible role of the octopaminergic neurons in the regulation of certain physiological function(s), active only from a late stage of embryonic development, is suggested.

Animals↗
PubMed1995

Octopamine immunoreactivity in the fruit fly Drosophila melanogaster.

Octopamine has been proposed as a neurotransmitter/modulator/hormone serving a variety of physiological functions in invertebrates. We have initiated a study of octopamine in the fruit fly Drosophila melanogaster, which provides an excellent system for genetic and molecular analysis of neuroactive molecules. As a first step, the distribution of octopamine immunoreactivity was studied by means of an octopamine-specific antiserum. We focused on the central nervous system (CNS) and on the innervation of the larval body wall muscles. The larval octopamine neuronal pattern was composed of prominent neurons along the midline of the ventral ganglion, whereas brain lobes were devoid of immunoreactive somata. However, intense immunoreactive neuropil was observed both in the ventral ganglion and in the brain lobes. Some of the immunoreactive neurons sent peripheral fibers that innervated most of the muscles of the larval body wall. Octopamine immunoreactivity was observed at neuromuscular junctions in all larval stages, being present in a well-defined subset of synaptic boutons, type II. Octopamine immunoreactivity in the adult CNS revealed many additional neurons compared to the larval CNS, indicating that at least a subset of adult octopamine neurons may differentiate during metamorphosis. Major octopamine-immunoreactive neuronal clusters and neuronal processes were observed in the subesophageal ganglion, deutocerebrum, and dorsal protocerebrum, and intense neuropil staining was detected primarily in the optic lobes and in the central complex.

Animals↗
PubMed1995

Evidence for cis interaction and cooperative signalling by the heat-stable antigen nectadrin (murine CD24) and the cell adhesion molecule L1 in neurons.

L1 is a transmembranal homophilic cell adhesion molecule of the immunoglobulin superfamily expressed by neural and lymphoid cells. The heat-stable antigen (HSA, murine CD24) nectadrin is a highly and heterogeneously glycosylated glycophosphatidylinositol-linked differentiation antigen of haematopoietic and neural cells. L1 and nectadrin have been shown to mediate cell adhesion and intracellular Ca2+ signals in neurons and B lymphoblasts, respectively. Here we show that nectadrin is co-expressed with L1 in murine cerebellar granule cell neurons and neuroblastoma N2A cells. Purified nectadrin bound to L1 with an apparent binding ratio of five nectadrin molecules to one L1 molecule at saturation. Binding between nectadrin and purified N-CAM was not observed. In co-capping experiments nectadrin co-redistributed with L1 and N-CAM. Since in these cells N-CAM and L1 cohere by cis-binding nectadrin appears to join the L1-N-CAM complex through binding to L1. Antibodies to each L1 and nectadrin evoked small increases in the intracellular Ca2+ concentration. However, when both antibodies were added together or in tandem to the cells, a strong intracellular Ca2+ signal was measured that was at least 6- and 10-fold stronger than the signal separately induced by L1 and nectadrin antibodies respectively. Such a cooperative effect was not observed in B lymphoblasts, using the same antibodies, or in neurons, using a combination of L1 and Thy-1 antibodies. Both the weak Ca2+ signal mediated by L1 alone and the enhanced signal jointly triggered by antibodies to L1 and nectadrin were inhibited by phorbol 12-myristate 13-acetate and were not significantly affected by Ni2+ and Cd2+ cations, suggesting that they are related to one another and involve recruitment of intracellular Ca2+. Nectadrin therefore appears to join a functional complex of neuronal adhesion molecules and to potentiate the signal transduction pathway of L1, possibly in response to neuron-neuron contact formation.

Animals↗
PubMed1995

Distribution and developmental expression of octopamine-immunoreactive neurons in the central nervous system of the leech.

Octopamine, a biogenic amine analogous to norepinephrine, plays an important role in the orchestration and modulation of invertebrate behavior. In the leech, the behavioral actions of octopamine have been demonstrated; however, identification of octopaminergic neurons had not been determined by using immunohistochemical techniques. Thus, we used an antibody highly specific to octopamine to examine the distribution of octopamine-immunoreactive neurons in the segmental ganglia of American and European medicinal leeches (Macrobdella decora and Hirudo medicinalis). One pair of octopamine-immunoreactive neurons was located in the dorsolateral ganglionic region of anterior ganglia 1-6 and posterior ganglia 15-21. No corresponding octopamine-immunoreactive neurons were found in midbody ganglia 7-14. Using Neutral Red staining in combination with intracellular Neurobiotin injections and octopamine immunostaining, we determined the identity of the dorsolateral octopamine-immunoreactive cells. The dorsolateral octopamine-immunoreactive neuron (the DLO) was not cell 21, the only previously reported Neutral Red staining neuron in the dorsolateral position. We also determined that the Leydig neuron was not octopamine immunoreactive in either of the two medicinal leech species. Octopamine immunostaining in the sex ganglia revealed hundreds of immunoreactive neurons in sexually mature leeches. Such neurons were not observed in juvenile leeches. The developmental time course of octopamine immunoreactivity in the dorsolateral octopamine-immunoreactive neurons was also investigated by staining embryonic Hirudo medicinalis. Octopamine expression occurred relatively late as compared with the detectable onset of serotonin expression. Octopamine expression in the dorsolateral octopamine-immunoreactive cells was not detectable at early to mid-embryonic stages, and must commence during late embryonic to early juvenile stages. The identification of octopamine-immunoreactive cells now sets the stage for further investigations into the functional role of octopamine in leech behavior and the development of behavior.

Animals↗
PubMed1995

Putative neurohemal areas in the peripheral nervous system of an insect, Gryllus bimaculatus, revealed by immunocytochemistry.

The morphology and position of putative neurohemal areas in the peripheral nervous system (ventral nerve cord and retrocerebral complex) of the cricket Gryllus bimaculatus are described. By using antisera to the amines dopamine, histamine, octopamine, and serotonin, and the neuropeptides crustacean cardioactive peptide, FMRFamide, leucokinin 1, and proctolin, an extensive system of varicose fibers has been detected throughout the nerves of all neuromeres, except for nerve 2 of the prothoracic ganglion. Immunoreactive varicose fibers occur mainly in a superficial position at the neurilemma, indicating neurosecretory storage and release of neuroactive compounds. The varicose fibers are projections from central or peripheral neurons that may extend over more than one segment. The peripheral fiber varicosities show segment-specific arrangements for each of the substances investigated. Immunoreactivity to histamine and octopamine is mainly found in the nerves of abdominal segments, whereas serotonin immunoreactivity is concentrated in subesophageal and terminal ganglion nerves. Immunoreactivity to FMRFamide and crustacean cardioactive peptide is widespread throughout all segments. Structures immunoreactive to leucokinin 1 are present in abdominal nerves, and proctolin immunostaining is found in the terminal ganglion and thoracic nerves. Codistribution of peripheral varicose fiber plexuses is regularly seen for amines and peptides, whereas the colocalization of substances in neurons has not been detected for any of the neuroactive compounds investigated. The varicose fiber system is regarded as complementary to the classical neurohemal organs.

Amines↗
PubMed1995

The role of proctolin in the antenna-heart beat acceleration of Periplaneta americana (L.).

So-called accessorial pulsatile circulatory organs in insects have developed during evolutionary reduction of the vascular system. As such a peripheral organ, the antenna-heart supplies the antennae with haemolymph. In the antenna-heart, the beat rhythm is generated by a myogenic automatism and is controlled neuronally. The electrical stimulation of the antenna-heart nerve produces an immediate heart beat inhibition brought about by octopamine release. Following this heart inhibition, an accelerating effect can be observed, which is based on a peptidergic mechanism. Proctolin-like material was found in both the dilator muscle and the antenna-heart nerve by means of immunocytochemistry. After HPLC-separation of antenna-heart tissue, proctolin was identified by sequencing the bioactive material and subsequent mass spectrometry. Proctolin proved to be extraordinarily effective producing stimulation of the antenna-heart beat rate up to 450%. The threshold concentration is 10(-10) mol.1(-1) and the value of the dissociation constant was fixed to KO = 1.3 x 10(-8) mol.1(-1). The chrono-trophic effect is caused by an increase in the steepness of the rising phase of pacemaker activity. The enhancement of the membrane resistance Rinput indicates a reduction of K(+)-conductance by proctolin. The PI-second messenger system is involved as well. Proctolin was identified and isolated from a real target organ. This fact and the physiological results furnished proof of peptidergic transmitter function of proctolin in the antenna-heart.

Animals↗
PubMed1994

Octopamine-like immunoreactivity in the brain and subesophageal ganglion of the honeybee.

The organization of putative octopaminergic pathways in the brain and subesophageal ganglion of the honeybee was investigated with a well-defined polyclonal antiserum against octopamine. Five prominent groups of just over 100 immunoreactive (IR) somata were found in the cerebral ganglion: Neurosecretory cells in the pars intercerebralis innervating the corpora cardiaca via NCC I, one cluster mediodorsal to the antennal lobe, one scattered on both sides of the midline of the protocerebrum, one between the lateral protocerebral lobes and the dorsal lobes, and a single soma on either side of the central body. With the exception of the pedunculi and beta-lobes of the mushroom bodies, varicose immunoreactive fibers penetrate all parts of the cerebral ganglion. Strong labelling was found in the central complex and the protocerebral bridge. Fine networks of labelled processes invade the antennal lobes, the calyces and a small part of the alpha-lobes of the mushroom bodies, the protocerebrum, and all three optic ganglia. In the subesophageal ganglion, one labelled cell body was found in the lateral soma layer of the mandibular segment. Each of the three neuromeres contains a group of six to ten somata in the ventral median parts. Most of the ventral median cells send their neurites dorsally through the midline tracts, whereas the neurites of a few cells follow the ventral cell body neurite tracts. Octopamine-IR was demonstrated in all neuropils that contain pathways for proboscis extension learning in honeybees. Because octopaminergic mechanisms seem to be involved in the behavioral plasticity of the proboscis extension reflex, our study provides anatomical data on the neurochemical organization of an appetitive learning paradigm.

Animals↗