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Biomedical subjects

R Huybrechts

Publications and source records attributed to R Huybrechts.

9 recordsLinked to original sources

Nucleotide sequence of a transactivating Bombyx mori nuclear polyhedrosis virus immediate early gene.

The open reading frame (ORF) of 1572 bp contained in the 3.8 kb ClaI fragment of BmNPV encodes a viral regulatory protein which transactivates the delayed early AcMNPV 39K gene. Transactivation is induced in uninfected cells following transfection with a plasmid containing only the ClaI fragment. Hitherto immediate early gene promoter activity of the included 631 bp leader sequence is evident since no other viral elements are needed for the transcription of the regulatory gene.

Animals

Immunohistochemical localisation of the hypertrehalosaemic hormone II (Cam-HrTH-II) and related peptides in the nervous system of Carausius morosus and Sarcophaga bullata.

A polyclonal antiserum was prepared against an N-terminal modified Cam-HrTH-II (Leu-Asn-Phe-...), one of the members of the large AKH/RPCH peptide family, first isolated from Carausius morosus. The localisation of this peptide was performed by means of immunocytochemical methods in the brain and corpora cardiaca-corpora allata complex of the stick insect, Carausius morosus and the grey fleshfly, Sarcophaga bullata. The distribution patterns of molecules reactive to the Cam-HrTH-II and the Lom-AKH-I antisera in both insect species were compared. In Carausius, both antisera reacted in the same cell bodies. In Sarcophaga, some neurons were stained by both, others only by one of the two antisera. By combining two different antisera, we demonstrated that there are no Lom-AKH-I-like molecules present in Carausius and that there must occur at least three different AKH-like molecules in the brain of Sarcophaga. One is similar to Cam-HrTH-II, the second to Lom-AKH-I and the third is an AKH/RPCH-like peptide, different from Lom-AKH-I and Cam-HrTH-II.

Amino Acid Sequence

A substance resembling somatomedin C in the American cockroach.

Material antigenically resembling somatomedin C (type I insulin-like growth factor, IGF-I) is demonstrated in the American cockroach Periplaneta americana by means of a monoclonal antibody immunoperoxidase technique. It was localized histochemically in neuronal cell somata and axonal fibers (probably interneurons) of the central nervous/neuroendocrine system and in 'endocrine-type' cells lining the midgut epithelium. The IGF-I-like substance is different from vertebrate insulin and also distinct from materials immunostained by different insulin antibodies in the brain and neurohaemal complex of this insect species. These findings are viewed in the light of recent reports on the presence and action of insulin-like chemicals in insects, and with respect to the existence of an insect brain-midgut system similar to the mammalian brain-gastroenteropancreatic system.

Animals

Coexistence of melanin-concentrating hormone and alpha-melanocyte-stimulating hormone immunoreactivities in the central nervous system of the locust, Locusta migratoria.

The distribution of melanin concentrating hormone (MCH) in the central nervous system of the locust Locusta migratoria was studied by the indirect immunofluorescence technique, using antibodies against salmon MCH. Most MCH-immunoreactive perikarya were found in the optic lobes at both sides of the brain, dorsally with respect to the lamina ganglionaris. The same neurons also contain alpha-melanocyte-stimulating hormone (alpha-MSH)-like material. In addition, a moderate number of MCH-like neurons, which were devoid of alpha-MSH-immunoreactive substances, was observed in the pars intercerebralis. Bright immunofluorescent fibers were visualized in various regions of the central nervous system of the locust: the optic lobes, the ocelli, the proto-and deuterocerebrum, the subesophageal connectives and the corpora cardiaca. In the ventral nerve cord and the subesophageal ganglion, where alpha-MSH-like cell bodies were encountered, MCH immunoreactive perikarya were absent and immunoreactive fibers were scarce. The coexistence of MCH and alpha-MSH-immunoreactive material within the same specific neurons might indicate an evolutionary relationship of both peptides.

Animals

Methionine-enkephalin immunoreactivity in the gonads and nervous system of two insect species: Locusta migratoria and Sarcophaga bullata.

Methionine(met)-enkephalin immunoreactivity as visualized by the peroxidase-antiperoxidase procedure, is present in spermatogonia, spermatocytes, spermatids, and young ovarian follicles of Locusta (panoistic type) and Sarcophaga (polytrophic type). Follicle cells and mature spermatozoa are always immunonegative as are locust vitellogenic follicles. In oocytes and in trophocytes, the met-enkephalin-like material first appears around the nucleus and is then dispersed throughout the cytoplasm. Later, it is present only in the periphery. In the ovary of both insects, no immunoreactivity is found with antisera against adrenocorticotrophic hormone, melanophore stimulating hormone, beta-endorphin, corticotropin releasing factor, or leucine-enkephalin. All these antisera yield a positive reaction when applied to the central nervous system as does the met-enkephalin antiserum. This study indicates that the met-enkephalin-like peptide may play a role in reproductive physiology.

Animals

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

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

NPY-like peptides occur in the nervous system and midgut of the migratory locust, Locusta migratoria and in the brain of the grey fleshfly, Sarcophaga bullata.

The distribution of the NPY-like substances in the nervous system and the midgut of the migratory locust, Locusta migratoria and in the brain of the grey fleshfly, Sarcophaga bullata was determined by immunocytochemistry using an antiserum directed against synthetic porcine NPY. The peroxidase-antiperoxidase procedure revealed that NPY immunoreactive cell bodies and nerve fibers were observed in the brain, optic lobes, corpora cardiaca, suboesophageal ganglion and ventral nerve cord of the locust and in the brain, optic lobes and suboesophageal ganglion of the fleshfly. In the locust midgut, numerous endocrine cells and nerve fibers penetrating the outer musculature contained NPY-like immunoreactivity. The concentrations of NPY immunoreactive material in acetic acid extracts of locust brain, optic lobes, thoracic ganglia, ovaries and midguts was measured using a specific radioimmunoassay technique. The dilution curves of the crude tissue extracts were parallel to the standard curve. The highest amount of NPY-like immunoreactivity was found in the locust ovary and midgut. Reverse-phase high-performance liquid chromatography (RP-HPLC) and radioimmunoassay were used to characterize the NPY-like substances in the locust brain and midgut. HPLC-analysis revealed that NPY-immunoreactivity in the locust brain eluted as three separate peaks. The major peak corresponded to a peptide less hydrophobic than synthetic porcine NPY. RP-HPLC analysis of midgut extracts revealed the presence of an additional NPY-immunoreactive peak which had a retention time similar to the porcine NPY standard. The present data show the existence of a widespread network of NPY immunoreactive neurons in the nervous system of the locust and the fleshfly. Characterization of the immunoreactive substances indicates that peptides similar but not identical to porcine NPY are present in the central nervous system and midgut of insects.

Animals