Biomedical subjects

J van Minnen

Publications and source records attributed to J van Minnen.

47 records · Page 3Linked to original sources

PubMed1990

Simultaneous detection of different mRNA sequences coding for neuropeptide hormones by double in situ hybridization using FITC- and biotin-labeled oligonucleotides.

Oligonucleotides labeled with FITC or biotin were applied for detection of specific mRNAs in microscopic preparations by in situ hybridization. The oligonucleotides were labeled with one FITC or biotin molecule at the 5' end or with a tail of biotin molecules at the 3' end. The target sequences were mRNAs coding for an ovulation hormone (CDCH) in the caudodorsal cells (CDC) of the pond snail Lymnaea stagnalis and a molluscan insulin-like peptide (MIP) in the light green cells (LGC) of the same organism. The hybridized oligonucleotides were detected either directly after the hybridization procedure by fluorescence microscopy or indirectly after an immunocytochemical procedure to visualize the biotin or FITC moiety. The results indicate that the detectability of the mRNA sequences is at least partially dependent on the accessibility of the target sequences for the immunocytochemical detection systems. The positive hybridization results obtained with oligonucleotides containing different labels enabled us to perform double hybridization experiments for simultaneous detection of CDCH and MIP mRNAs in one tissue section. Using FITC- and biotin-labeled oligonucleotides, we also demonstrated simultaneously different sequences on the same mRNA molecule.

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PubMed1989

Expression of the egg-laying hormone genes in peripheral neurons and exocrine cells in the reproductive tract of the mollusc Lymnaea stagnalis.

The neuroendocrine caudodorsal cells play an important role in the control of reproduction in Lymnaea stagnalis. These neurons produce at least nine neuropeptides which are encoded by caudodorsal cell hormone-I and -II genes. The role of some of these peptides in the control of reproduction has been established. The present study demonstrates that the transcription and translation of the caudodorsal cell hormone genes also proceeds abundantly in the reproductive tract of this hermaphroditic animal. In the female part of the reproductive tract neurons were found to express gene I. These neurons are most likely involved in the control of transport of the eggs and egg-masses and in the regulation of secretory activity from the female accessory sex glands. In the male part of the reproductive tract exocrine secretory cells express gene I or gene II. The gene products are secreted into the male duct and transferred to the female copulant during copulation. Furthermore, putative sensory neurons in the skin were found to express gene I. The results indicate that in L. stagnalis the complex process of reproduction is regulated--at least in part--by a set of neuropeptides which are encoded by a small multigene family, viz. the caudodorsal cell gene family.

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PubMed1989

Central and peripheral expression of genes coding for egg-laying inducing and insulin-related peptides in a snail.

Egg laying in the hermaphrodite freshwater snail Lymnaea stagnalis is a highly complex activity, including a series of internal activities (ovulation, egg and egg mass formation) which are closely correlated to a pattern of behaviours (alteration of locomotion and feeding, specific postures, oviposition). In this snail egg laying is induced by the neuroendocrine caudodorsal cells (CDCs), consisting of two homogeneous clusters at a total of 100 neurons. At egg laying these neurons release their products during a 60 min period of firing. The genes coding for these products have been cloned and characterized. There are two genes, CDCH-I and -II. Each gene codes for 12 peptides; one of these is the ovulation hormone (CDCH). The genes display over 90% homology. The most striking difference is a 17 bp deletion near the carboxy-terminal region. With immunocytochemistry and in situ hybridization both CDCH genes appeared to be expressed in the CDC and in paired groups of ectopic CDC-like neurons in the pleural ganglia, while a group of small neurons in the cerebral ganglia expresses the CDCH-I gene only. In addition, a widespread expression of the CDCH genes has been demonstrated in peripheral tissues. In the female part of the reproductive tract neurons were found to express the CDCH-I gene. In the male part of the tract exocrine secretory cells express the CDCH-I or -II gene. The gene products are secreted into the male tract and transferred to the female partner during copulation. Finally, sensory neurons in the head skin and mantle edge were found to express the CDCH-I gene. The presence of insulin-related peptides has been demonstrated in the brain as well as the digestive system of Lymnaea. The brain insulin-related peptides are produced in 4 groups of 50 giant neurons each (Light green cells, LGC). These neurons are involved in various physiological activities, related to body growth and glycogen metabolism. The major gene products expressed in the LGC have been cloned and characterized. It appeared that the predicted proteins represent three types of insulin-related molecules (MIP, molluscan insulin-related peptide). In these MIPs, those elements important in the determination of the tertiary structure, have been conserved. The MIP of the digestive system has been characterized up to now only at the peptide level. The snail gut MIP is more hydrophobic compared to bovine insulin. Cells containing MIP have been identified immunocytochemically in the gut epithelium.

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PubMed1988

Localization of ovulation hormone-like neuropeptide in the central nervous system of the snail Lymnaea stagnalis by means of immunocytochemistry and in situ hybridization.

The caudo-dorsal cells (CDC) in the cerebral ganglia of the pond snail Lymnaea stagnalis synthesize the 36-amino acid ovulation hormone (CDCH). We have used immuno-cytochemistry and in situ hybridization to reveal the localization of neurons and axons containing CDCH-like material. A monoclonal antibody to a fragment of CDCH and a cDNA probe encoding CDCH reacted with the CDC-system, with specific cell groups in the cerebral and pleural ganglia, and with individually occurring neurons throughout the central nervous system. The cells in the pleural ganglia, which were found in about 50% of the preparations studied, are considered as "ectopic" CDC. They are morphologically similar to CDC in their somal dimensions and axonal organization. By means of immuno-electron microscopy it was shown that these neurons contain secretory vesicles that are similar to those of the CDC. The neurons of the bilateral groups occurring in the cerebral ganglia in addition to the CDC are smaller and more intensely stained than the CDC. Axons of these small neurons probably have varicosities located on the CDC axons in the neuropil of the cerebral ganglion, indicating synaptic contacts. Two major axon tracts could be followed from (or toward) the neuropil of the cerebral ganglion. One tract runs from the cerebral ganglion via the pleural and parietal ganglia to the visceral ganglion, giving off branches to most nerves emanating from these ganglia. The other tract could be traced through the cerebro-pedal connective to the pedal ganglia. Only in the right pedal ganglion was extensive axonal branching observed.(ABSTRACT TRUNCATED AT 250 WORDS)

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PubMed1987

Demonstration of substances immunologically related to the identified arthropod neuropeptides AKH/RPCH in the CNS of several invertebrate species.

The occurrence of adipokinetic hormone (AKH) and red pigment concentrating hormone (RPCH)-like neuropeptides in the central nervous system of different invertebrate species other than insects was investigated immunocytochemically with polyclonal antisera to N- and C-terminal regions of the AKH molecule. Substances reacting with the C-terminal specific antiserum code 241 were present in neurons of the pond snail, Lymnaea stagnalis, the sowbug Porcellio scaber, the centiped Lithobius forficatus and the crayfish Astacus leptodactylus. Substances revealed by the N-terminal specific antiserum code 433 were demonstrated in the latter two species. The distribution of immunoreactive substances in neuropile areas of several ganglia suggests that these substances act as neurotransmitter/neuromodulator. Their presence in neurohemal organs such as the periphery of the pedal and visceral nerves in the pond snail and in the sinus gland of the crayfish suggests a neurohormonal role in these species.

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PubMed1984

Galactogen synthesis-stimulating factor in the slug, Limax maximus: cellular localization and partial purification.

Galactogen synthesis in the albumen gland of the slug Limax maximus is under direct endocrine control [J. van Minnen, J. Wijdenes, and P. G. Sokolove (1983) Gen. Comp. Endocrinol. 49, 307-314]. The present results show that dorsal body cells (DBC) present throughout the connective tissue around the circumesophageal brain are the major cellular source of a galactogen synthesis-stimulating factor (GAL-SF). The amount of GAL-SF varies during sexual maturation of L. maximus (a protandrous hermaphrodite): it is low when animals are immature and reaches a peak in the late female phase when the animals prepare to lay eggs. The morphology of DBC also changes during maturation. DBC are small and release little secretory product in immature and early male-phase animals; release is higher and DBC are larger in late female-phase animals. Gel filtration of homogenates of cerebral ganglia on Sephadex G75 yields one peak of GAL-SF activity with an estimated Mr of 4000-7000 Da. Subsequent anion exchange chromatography showed that GAL-SF could be eluted with a linear NaCl gradient; GAL-SF was found in fractions with a NaCl concentration between 0.04 and 0.07 M.

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PubMed1983

Endocrine control of galactogen synthesis in the albumen gland of the slug, Limax maximus.

An in vitro method for culturing Limax maximus albumen glands is described in which the biosynthetic activity of the slug albumen gland was monitored by measuring the incorporation of [14C]glucose into galactogen. Homogenates of the central nervous system were shown to cause a 3.5- to 12-fold increase in galactogen synthesis in albumen gland explants as compared to controls. The major sources of the galactogen-synthesis stimulating factor (Gal-SF) were found to be the cerebral ganglia and their surrounding connective tissue. Gal-SF was demonstrated to be peptidase sensitive and heat labile suggesting that it is probably a polypeptide. Autoradiographs of semithin araldite sections supported the incorporation data: in albumen gland explants cultured with cerebral ganglion homogenate considerably more label was found over secretory granules than in control-cultured explants. The possible cellular source of Gal-SF is discussed in relation to its possible origin in other investigated pulmonates.

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PubMed1980

A comparative study on neurosecretion demonstrated by the alcian blue-alcian yellow technique in three terrestrial pulmonates (Stylommatophora).

Neurosecretion was studied in the central ganglia of three terrestrial pulmonates (Deroceras reticulatum, Arion hortensis, and Helix aspersa) with the alcian blue-alcian yellow (AB/AY) staining method. A number of neurosecretory cell types were distinguished in all three species (D. reticulatum: 9, A. hortensis: 11, H. aspersa: 13) With AB/AY the cells stain different shades of green and yellow. The histochemical differences reflect ultrastructural differences: the cell types contain elementary granules of different size and appearance. Some cell types form distinct groups of up to 50 cells, while others occur in smaller groups or as single cells. Neurosecretory axons could only be traced in a few cases to the probable neurohaemal areas. Based on the following criteria: (1) staining properties of cells (2) size of cells, (3) position of cells in the ganglia, it appears that the neurosecretory systems of the three species show great similarity, although some differences are apparent.

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PubMed1980

Photoperiod-dependent neural control of the activity of the neurosecretory canopy cell in the lateral lobes of the cerebral ganglia of the freshwater pulmonate Lymnaea stagnalis (L.).

In the lateral lobes of the cerebral ganglia of the pulmonate freshwater snail Lymnaea stagnalis (L.) one large neurosecretory canopy cell (CC) and two droplet cells (DC) are present. With the ganglion-transplantation technique and with quantitative electron microscopy it is shown that photoperiodic stimuli control the neurosecretory activity of the CC via a stimulatory extrinsic neural pathway. The eyes do not play an important role in this regulatory process. The neurosecretory activity of the DC is not affected by differences in photoperiod. The significance of the photoperiodic control of CC activity is discussed in relation to the pathway for the endocrine control of reproduction which is known to be photoperiod-dependent.

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PubMed1979

An ultrastructural study of the neurosecretory canopy cell of the pond snail Lymnaea stagnalis (L.), with the use of the horseradish peroxidase tracer technique.

The paired, electrotonically coupled neurosecretory Canopy Cells (CC) of the pond snail Lymnaea stagnalis were microiontophoretically injected with horseradish peroxidase (HRP). Whole mount preparations and ultrathin sections of injected CC were studied to describe in detail the morphology of the CC, their axon tracts and neurohaemal areas. The CC release their secretory product at three different sites, viz. from the soma and from axon terminals in the intercerebral commissure and in the median lip nerve. Neural control over the CC occurs by few synapses found exclusively on the CC axon, not on the cell body. One type of "en passant" synapse was identified. Two morphologically distinct types of synapselike structures (SLS) are numerous. The site of electrotonic coupling between the two CC is most probably located in the cerebral commissure. Serial sectioning showed that the axons contact each other over more than 130 micrometers. The contact is very intimate and the axon membranes interdigitate in a complex manner. Gap junctions, which are often described as the sites of electrotonic coupling, were not found.

Animals↗

PubMedDate not supplied

Production and exocrine secretion of LHRH-like material by the male rat reproductive tract.

Luteinizing hormone-releasing hormone (LHRH)-like immunoreactivity was localized in the male reproductive system of the rat. Epithelial cells of the epididymus, seminal vesicles and coagulation gland showed a strong reaction to anti-LHRH serum. Also the epithelia of the ductus deferens and the prostate gland appeared to be immunoreactive, albeit to a lesser extent. The LHRH-like substances are most likely secreted into the male tract, as can be concluded from the observation that the secretion product in the lumina of the seminal vesicles, coagulation gland and prostate gland was also immunopositive. The functional significance of these phenomena is discussed. No immunostaining was obtained with antisera to FSH, LH or beta-hCG.

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