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Pancreatic enzyme storage and secretion were studied in rats treated twice daily with SC injections (5 μg/kg) of CCK-8 3, 7, and 15 days. Isolated zymogen granules were analyzed by flow cytometry to determine their FSC (forward scatter), SSC (side scatter), and amylase and trypsinogen contents. DNA content, pancreatic weight, and both basal and stimulated pancreatic secretion under IV CCK infusion...
We have isolated TDVDHVFLRFamide (DMS), FDDYGHMRFamide (DSK), and DPKQDFMRFamide from Drosophila melanogaster. These peptides, structurally related by a common C-terminus -XRFamide, where X = L or M, are encoded by three different genes. To determine cellular expression, we have generated antisera to multiple antigenic peptides and performed double-label immunofluorescence using antisera raised...
The relationship between gallbladder (GB) bile concentration and motility was studied in conscious dogs. The 12-h GB bile concentrations between meals could be divided into three periods: diluting, minimum, and concentrating periods. During the diluting period, inhibition of GB contractions by a CCK A receptor antagonist, atropine or hexamethonium, resulted in concentration of GB bile, whereas...
Valorphin, an endogenous opioid-like hemoglobin fragment, is cytotoxic for L929 and K562 tumor cells in 10 -7 -10 -13 M concentration range. Because cytolytic effects induced by valorphin in K562 cells are inhibited by naloxone, opioid receptors should be involved in induction of valorphin-mediated tumor cell death. Three distinct cytolytic processes, differing in the...
Flavostatin, a novel disintegrin purified from the venom of Trimeresurus flavoviridis, consists of 68 amino acids, including an Arg-Gly-Asp sequence and 12 Cys residues at positions highly conserved among disintegrins. The N-terminal sequence of flavostatin was identical to those of triflavin and flavoridin, previously reported disintegrins from the Trimeresurus flavoviridis venom. Differences among...
The myoinhibitory peptide leucomyosuppressin (LMS) (pQDVDHVFLRFamide) has been identified and characterized at the molecular level in the cockroachDiploptera punctata through analysis of the organization of both brain cDNA and genomic DNA. Processing of the precursor predicted from DNA sequence would release a single LMS peptide. The organization of the precursor appears to be conserved in other...
Plasma endothelin-1 was measured around the clock in 72 subjects. Cosinor methods were used to assess circadian and other recurrent variation and trends, that is, the time structure (chronome) of this peptide. Multifactorial analyses of variance and linear regressions assessed chronome alterations associated with different risk factors: diabetes, obesity, high cholesterol, high blood pressure, vascular...
The first peptide identified in locusts was adipokinetic hormone I (AKH-I), a neurohormone mobilizing lipids from the fat body. No other locust peptides were isolated until 1985. From then on peptide identification started to boom at such a tremendously fast rate that even specialists in the field could hardly keep track. At this moment the total number of different insect neuropeptide sequences...
The uptake of a nonmetabolizable derivative of glucose, [ 3 H]2-deoxy-dglucose, was examined in isolated slow (soleus) and fast (extensor digitorum longus, EDL) muscles of adult mice. An analogue of β-endorphin (28-31), Ac-Lys-d-Lys-Sar-Glu, which is stable to proteolytic digestion, enhanced the uptake of glucose into the slow and fast muscles. The muscles of male mice were more sensitive...
Responses to a newly synthesized human adrenomedullin (hADM) analog, hADM(16-31), were investigated in the rat and cat. Unlike the full-sequence peptide, which has potent hypotensive activity, hADM(16-31) had pressor activity in the rat but not in the cat. Injection of hADM(16-31) in doses of 10-300 nmol/kg IV induced dose-dependent increases in systemic arterial pressure in the rat, and the peptide...
Analogues of the insect kinin family in which the Xaa 2 residue of the C-terminal pentapeptide core sequence Phe-Xaa 1 -Xaa 2 -Trp-Gly-NH 2 (Xaa 1 = Asn, His, Phe, Ser, or Tyr; Xaa 2 = Ala, Ser, or Pro) is replaced with sterically hindered aminoisobutyric acid (Aib) prove to be resistant to hydrolysis by housefly (Musca domestica) angiotensin-converting...
STKR is a 4118 bp clone from a stable fly, Stomoxys calcitrans, cDNA library which encodes a protein with significant amino acid identity to tachykinin-like peptide receptors. Ribonuclease protection assays and RT-PCR were utilized to examine the transcriptional expression of STKR from various life stages of the stable fly. STKR expression was detectable in all stages, but was most abundant in isolated...
Incubation of Dip-AST 7 (APSGAQRLYGFGLa) or Dip-AST 9 (GDGRLYAFGLa) (5 μM) with hemolymph for 30 min results in cleavage by a putative endopeptidase, yielding the C-terminal hexapeptide. This metabolic product is subsequently cleaved by an amastatin-sensitive aminopeptidase to yield the C-terminal pentapeptide, as treatment with the competitive aminoexopeptidase inhibitor, amastatin, results in a...
FMRFamide-like immunoreactivity was localized immunohistochemically in the central and stomatogastric nervous systems, visceral tissues, and the neurohemal corpora cardiaca, transverse, and segmental nerves. Each of these neurohemal areas contains one morphologically distinct type of immunoreactive neurosecretory granule. The hemolymph level of FMRFamide-like peptides, quantified by RIA, is higher...
A radioimmunoassay was developed using 125 I-labeled-[Tyr 0 ]Locusta-DH and polyclonal antibodies raised against Locusta-DH (29-46). The assay had a detection limit of 50 pM, and displayed limited cross-reactivity for other CRF-related peptides, but not for unrelated peptides. About 60% of the total immunoreactive material in locust hemolymph was attributable to Locusta-DH...
The presence in insect tissues of peptides with structural similarities to angiotensin I and to bradykinin, the two best known substrates of mammalian angiotensin-converting enzyme, has not been reported. As part of our study to identify potential substrates for insect angiotensin-converting enzyme, we have investigated the susceptibility of a number of known insect peptide hormones and neurotransmitters...
Receptors for neuropeptide Y (NPY) are widely distributed throughout the mammalian brain. Using indirect labeling methods, the human brain was reported to contain predominantly the Y 2 receptor subtype, whereas the rat brain contains a mixture of Y 1 and Y 2 receptor subtypes. To more accurately assess NPY receptors in the human brain, we used the Y 1 - and Y ...
Four isomers of the Somatostatin analogue H-D-Phe-Cys-Tyr-D-Trp-Arg-Thr-Pen-Thr-NH 2 (CTAP) were made with β-MePhe in position 1 and assayed for opioid binding in rat brain, biological activity in MVD and GPI bioassays, and antinociception in mouse warm-water tail flick assays. These analogues displayed varying potencies and biological activities including: simultaneous δ receptor agonism/μ...
We have isolated seven tachykinin-related peptides (TRPs) from an acidic extract of 1000 brains of the cockroach Leucophaea maderae. Four different reversed-phase high performance liquid chromatography (RP-HPLC) column systems were required to obtain pure peptides. During the purification the fractions were monitored in a radioimmunoassay (RIA) with an antiserum to the locust TRP locustatachykinin...
The locust oviduct bioassay was used to assess a variety of proctolin analogues as possible agonists and antagonists of the peptide proctolin. Both [α-methyl-l-tyrosine 2 ]proctolin and [N-methyl-l-tyrosine 2 ]proctolin were agonists of proctolin with thresholds of 5 10 -9 M. Interestingly, at these threshold doses the analogues were antagonists when applied along with...
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