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Biomedical subjects

A De Loof

Publications and source records attributed to A De Loof.

At least 145 records · Page 8Linked to original sources
PubMed1988

The presence of a pregnenolone-binding factor in the copulatory organ of the migratory locust, Locusta migratoria migratorioides R. & F.

The presence of binding sites for nonecdysteroid steroids was investigated in the cytosol of several tissues of the migratory locust, Locusta migratoria migratorioides. Binding of androgens was not observed. Most tissues, however, showed nonsaturable binding of estrogens and in some tissues saturable progestin binding could be demonstrated. A pregnenolone binder, that was found to be present in the male copulatory organ, was further studied. It showed a dissociation constant of 4.4 (+/- 1.6) X 10(-8) M. This is the first report of a nonecdysteroid steroid-binding factor in an insect tissue.

Animals↗
PubMed1988

Biological, chromatographical, and radioimmunological evidence for a melanotropin-like peptide in the central nervous system of Locusta migratoria.

Our recent immunocytochemical study has demonstrated the existence of alpha-melanocyte stimulating hormone (alpha-MSH)-like material in the locust central nervous system. The aim of the present study was to further characterize alpha-MSH in the locust brain by its biological effect on frog skin and by high-pressure liquid chromatography in combination with radioimmunological and biological detection methods. Parallel radioimmunoassay (RIA) curves of crude nervous tissue extracts coupled with bioactivity in a very specific bioassay suggest similarity between the locust alpha-MSH-like substance and synthetic alpha-MSH. The highest concentration of alpha-MSH immunoreactive material in the central nervous system was found in the optic lobes, where alpha-MSH immunoreactive cell bodies are localized, as was previously shown by immunocytochemistry. High concentrations of alpha-MSH immunoreactive material were also detected in the thoracic ganglia of the locust ventral nervous system. The application of locust brain extracts to gel permeation HPLC resulted in a similar elution profile of the bioactive and immunologically active substances, both coeluting with synthetic alpha-MSH. As is the case for vertebrate brain material, reverse-phase HPLC revealed four alpha-MSH immunoreactive peaks. One of the peaks coelutes with monoacetyl-alpha-MSH and other RIA-positive material elutes at times close (but not identical) to the methionine sulfoxide forms of alpha-MSH. Peak three, however, elutes in a very different position from desacetyl-alpha-MSH. Peaks are absent in the position of desacetyl-alpha-MSH. Similarity between the locust alpha-MSH-related substance and authentic alpha-MSH is discussed.

Animals↗
PubMed1988

Concanavalin A reactivity of vitellogenin and yolk proteins of the threespined stickleback Gasterosteus aculeatus (Teleostei).

1. Concanavalin A (con A) reactive proteins have been detected in the plasma and ovaries of the oestradiol treated Gasterosteus aculeatus. 2. Concanavalin A-horseradish peroxidase (HRP) technique applied on nitrocellulose membranes reveals that vitellogenin (Vg) is the only mannose and glucose rich glycoprotein present in the plasma of oestradiol treated sticklebacks. Stickleback Vg can be purified by con A-Sepharose chromatography. 3. Con A reactivity in the ovary changes in the course of development of the oocytes. First, the yolk vesicles, which are synthesized by the oocyte itself, become con A positive. Later, the yolk granules, which contain vitellogenin synthesized in the liver and taken up from the plasma, show a clear affinity for con A. Con A staining disappears when mannopyranoside is added. 4. No con A staining is found in the periodic acid/Schiff staining chorion.

Animals↗
PubMed1987

Immunocytochemical demonstration of proopiomelanocortin- and other opioid-related substances and a CRF-like peptide in the gut of the american cockroach, Periplaneta americana L.

Using the peroxidase-antiperoxidase technique, we showed the presence of peptides which are immunologically resembling mammalian corticotropin releasing hormone (CRF)-, adrenocorticotropic hormone (ACTH)-, beta-endorphin (beta-END)-, alpha-melanocyte stimulating hormone (alpha-MSH)-, methionine-enkephalin (met-ENK)- and leucine enkephalin (leu-ENK)- like immunoreactivity in hundreds to thousands of endocrine cells and nerve fibers in the midgut of the American cockroach Periplaneta americana. In the cockroach hindgut no immunoreactive cell bodies could be observed, although nerve fibers were clearly noticed to be recognized by antisera to CRF, ACTH1-24, ACTH11-24 and beta-END. Nothing is exactly known as to the function(s) of the demonstrated materials, but one can speculate that these numerous immunoreactive cells, might have important paracrine and/or endocrine functions in the insect physiology.

Adrenocorticotropic Hormone↗
PubMed1987

Localization of melanotropin-like peptides in the central nervous system of two insect species, the migratory locust, Locusta migratoria, and the fleshfly, Sarcophaga bullata.

By use of well characterized antisera in the peroxidase-antiperoxidase method, we were able to demonstrate alpha MSH and beta MSH immunoreactive cells and nerve fibres within the nervous system of adults and larvae of Locusta migratoria and 3-, 5- and 8-day-old adult Sarcophaga bullata. In neither of these insect species, any immunoreaction was obtained with a gamma 3MSH-antiserum. Double immunohistochemical stainings revealed that alpha MSH-like and beta MSH-like substances are located in different cells. These cells show no immunoreactivity to a number of antisera against other POMC-derivatives (anti-beta lipotropin, anti beta endorphin, anti-ACTH1-24); thus they appear to contain alpha MSH- or beta MSH-like material in a specific way. The function of the immunologically detected peptides remains to be demonstrated. The distribution of the immunoreactive material suggests that, like in amphibians and other lower vertebrates, the synthesis or release of melanotropins might be under the influence of external stimuli. The present observations support the recently developed concept that even some of the smallest neuropeptides, the melanotropins, have been highly conserved during a long period of evolution.

Animals↗
PubMed1987

A comparative study of some properties of vitellogenin (Vg) and yolk proteins in a number of freshwater and marine teleost fishes.

1. Injection of oestradiol results in induction of vitellogenic proteins in the plasma of stickleback, sea bass, sea horse, pipe-fish, rainbow trout and roach. 2. Immunoblotting after SDS-PAA gradient gel electrophoresis of plasma proteins of stickleback, sea bass and rainbow trout reveals that vitellogenins from these species share common antigenic determinants. 3. Yolk proteins are recognized by antivitellogenin antisera which shows that proteolytical cleavage does not destroy all antigenic determinants. 4. Vitellogenic proteins are present only in the yolk granules and near the theca cell layer as is shown by immunocytochemical methods.

Animals↗
PubMed1987

Study of the metabolism of steroids in larvae of the fleshfly Sarcophaga bullata.

1. Larvae of the fleshfly Sarcophaga bullata were injected with several 3H C21 and C19 steroids. After different incubation times, the larvae were homogenized and the metabolites were extracted and fractionated by Sephadex LH 20-, paper- and thin-layer chromatography. The chromatographic mobility of the labeled zones was compared with that of standard steroids. 2. Progesterone and 17 alpha-hydroxypregnenolone were metabolized to 17 alpha-hydroxyprogesterone. Androstenedione, 17 alpha-hydroxyprogesterone and dehydroepiandrosterone were converted to testosterone. Transformation of pregnenolone to progesterone or 17 alpha-hydroxypregnenolone was not observed. 3. C21 or C19 steroid formation from cholesterol could not be demonstrated. 4. Sixteen metabolites, different from all our standard substances have been found. Their structure remains to be elucidated.

Androgens↗
PubMed1987

Distribution of yolk polypeptides in the follicle cells during the differentiation of the follicular epithelium in Sarcophaga bullata egg follicles.

In S. bullata, the ovaries contribute to the synthesis of yolk polypeptides. A specific antiserum for yolk polypeptides was used to visualize the presence of yolk polypeptides in the follicle cells during their differentiation. After vitellogenesis has started, all follicle cells contain yolk polypeptides. The squamous follicle cells covering the nurse cells and the border cells lose yolk polypeptides before mid-vitellogenesis, whereas the follicle cells over the oocyte contain yolk polypeptides until after late vitellogenesis. All follicle cells are immunonegative afterwards. In vitro translation of poly(A)+ RNA demonstrated that the presence of yolk polypeptide mRNA correlates well with follicle cell immunopositivity for yolk polypeptides. This suggests that the follicle cells synthesize the ovarian yolk polypeptides. Differences in cellular and nuclear morphology, total and poly(A)+ RNA synthesis and the rate of yolk polypeptide synthesis were shown to be correlated with the presence or absence of yolk polypeptides in the differentiating follicular epithelium. The possible relationship between these different aspects of follicle cell differentiation, follicle cell polyploidy and the extracellular current pattern around follicles are discussed.

Animals↗
PubMed1986

Polyploidization and localisation of poly(A)+ RNA in the different cell types of the vitellogenic meroistic ovary of the fleshfly, Sarcophaga bullata.

The degree of polyploidization, the level of transcriptional activity and the volume of the different cell types present in the meroistic ovary of Sarcophaga bullata were measured during different vitellogenic stages. The nurse cells and the germinal vesicle exhibited very pronounced differences with regard to DNA content and mRNA synthesis, even though they are genetically identical. During the 4C stage (late vitellogenesis), we observed different degrees of polyploidy in follicle cells adjacent to the oocyte and those surrounding the nurse cells. Although the chromatin of the germinal vesicle is condensed into a karyosome, in situ hybridisation revealed the presence of transcriptional activity. The volume of the germinal vesicle, which contains only 4C DNA, is big enough to contain 2048C DNA. The meroistic ovary is a highly polarized differentiating system. Our results are discussed in the light of the fact that the polytrophic ovary is a miniature electrophoresis chamber.

Animals↗
PubMed1986

Identification of nonecdysteroid steroids in hemolymph of both male and female Astacus leptodactylus (Crustacea) by gas chromatography-mass spectrometry.

The O-pentafluorobenzyloxime (OPFB)-heptafluorobutyrylester (HFB) derivatives of hemolymph steroid extracts from both male and female Astacus leptodactylus were subjected to negative ion chemical ionization and capillary gas chromatography-mass spectrometry (NCI/GC-MS). Five nonecdysteroid steroids, namely pregnenolone, 17 alpha-hydroxypregnenolone, testosterone, cholesterol, and 6 beta-hydroxyprogesterone were identified. With selected ion monitoring (SIM), indications were found for the presence of four more steroids: androstenedione, 5 alpha-dihydrotestosterone, 11-ketotestosterone, and 11 beta-hydroxytestosterone. The presence of 6 beta-hydroxyprogesterone could only be demonstrated in the female hemolymph. With the technique used, dehydroepiandrosterone, progesterone, 17 alpha-hydroxyprogesterone, and estrogens could not be detected in male or female hemolymph extracts.

17-alpha-Hydroxypregnenolone↗
PubMed1986

A new alternative for simultaneous immunohistochemical screening of 96 hybridoma clones for tissue-specific antibody productions selects a monoclonal antibody to insect corpus cardiacum.

In the current search for the elucidation of the true structure of hitherto unidentified 'new' insect neuropeptides we designed a novel screening method to facilitate the primary detection of neurone-specific antibody secreting mouse-mouse hybridoma clones obtained after immunization with neuronal tissue homogenates. The present procedure is principally adapted from a conventional immunohistological test and enables one to rapidly screen 96 (and even more) clones at one time for potential secretion of specific antibodies to different tissue compounds, without the necessity of having a purified antigen. It has proved to be sensitive, rapid, practical and reproducible. As such it promises to be very useful to discriminate amongst the wide range of antibodies to various kinds of materials produced by hybridomas by detecting monoclonal antibodies directed against factors contained in well-defined tissues in which one is interested. This paper also reports the successful application of this method to a primary screening of clones producing murine monoclonal antibodies to substances of insect corpora cardiaca (CC), after immunization with crude antigen preparations.

Animals↗
PubMed1985

Distinct localization of FMRFamide- and bovine pancreatic polypeptide-like material in the brain, retrocerebral complex and suboesophageal ganglion of the cockroach Periplaneta americana L.

One bovine pancreatic polypeptide (BPP) antiserum and two FMRFamide antisera were applied in the peroxidase-antiperoxidase (PAP) immunohistochemical technique on a complete series of sections of brains, suboesophageal ganglia (SOG), corpora cardiaca (CC) and corpora allata of Periplaneta americana L. Double immunohistochemical staining demonstrated that the same perikarya and processes were stained by both the BPP and FMRFamide antisera. This was caused by cross-reaction of the BPP and FMRFamide antisera with common antigenic determinants as was shown by a number of solid-phase absorptions. Application of a third FMRFamide antiserum, which was especially selected for its inability to react with bovine and avian pancreatic polypeptide, showed that more than half of the structures that were stained with the 'unspecific' BPP and FMRFamide antisera, contained material which was genuinely FMRFamide-like. This peptide material was located in cerebral neuronal structures, in the SOG, in the storage site of the CC and in numerous nerve fibres throughout the neuropile regions, which suggested a neurotransmitter/modulator as well as a neurohormonal role. The FMRFamide-like peptide was also found to be present in the same brain sites as an adipokinetic hormone-like peptide, but double labelling revealed that these two substances were never located in the same perikarya or fibres.

Animals↗
PubMed1985

Immunocytochemical localization of a methionine-enkephalin-resembling neuropeptide in the central nervous system of the American cockroach, Periplaneta americana L.

Using the peroxidase antiperoxidase immunocytochemical method, we were able to demonstrate within the brain and retrocerebral complex of Periplaneta americana several neuronal structures which were very specifically stained with an anti-methionine-enkephalin antiserum. From the precise localization of this immunoreactive material some speculations about its possible functions could be derived, such as a neurotransmitter- or neuromodulatorlike function and/or a neurohormonal role. These data present new evidence for the recently developed concept that opiate peptides, identical or related to those found in higher species, occur also in invertebrates.

Animals↗
PubMed1985

Substance P-like immunoreactivity in the central nervous system of the blattarian insect Periplaneta americana L. revealed by a monoclonal antibody.

Brains, retrocerebral complexes and frontal and suboesophageal ganglia of adult American cockroaches, Periplaneta americana, were immunohistochemically investigated with a specific monoclonal antibody (McAb) directed against a well characterized antigenic determinant, namely the COOH terminus of the endecapeptide substance P (SP). This resulted in the detection of several neurons and nerve fibres containing a substance antigenically closely related to this typically vertebrate neuropeptide. No difference in staining pattern could be observed between male and female insects. Related to the age of the adult specimens, however, a slight quantitative difference in SP immunoreactivity seems to occur, which probably might have functional implications. The SP-like peptide demonstrated in this study appears to be located in different neuronal structures than the ones that we earlier described as containing ACTH-, CRF-, OT-, AVP-, NP I-, NP II-, BPP-, FMRFamide-, AKH-, met-ENK-, FSH-, LH- and LHRF-like material (Verhaert et al. 1984a, b, 1985; Verhaert and De Loof 1985a, b).

Animals↗
PubMed1984

Localization of CRF immunoreactivity in the central nervous system of three vertebrate and one insect species.

By use of a specific antiserum against synthetic ovine corticotropin-releasing factor (CRF) in the peroxidase-antiperoxidase (PAP) immunocytochemical procedure (Vandesande and Dierickx 1976), CRF-like antigenic determinants were demonstrated in the central nervous system of a human fetus, the Wistar rat, the frog Rana ridibunda, and the American cockroach Periplaneta americana. The immunoreactive CRF-producing cells occur mainly in the nucleus paraventricularis of the rat, while in Rana ridibunda these cells occur in the nucleus praeopticus. Immunoreactive CRF-containing fibres were also visualized. Very clear CRF-immunoreactive products were observed in the brain as well as the corpora cardiaca (CC) and corpora allata (CA) of the cockroach Periplaneta americana. ACTH-immunoreactivity was also demonstrated in the brain-CC-CA complex of this insect. Double immunohistochemical staining (Vandesande 1983) also revealed that both the CRF- and ACTH-like substances occur in different neurosecretory neurons and nerve fibres. These results suggest that the antigenic determinants of CRF are very similar in vertebrates and insects bespeaking their very long evolutionary history.

Adrenocorticotropic Hormone↗