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T K Hayes

Publications and source records attributed to T K Hayes.

45 records · Page 3Linked to original sources

Neuroendocrine and juvenile hormone effects on fat body protein synthesis during the reproductive cycle in female Blaberus discoidalis cockroaches.

Ovarian protein content and fat body protein synthesis were measured during the first gonotrophic cycle in virgin female Blaberus discoidalis cockroaches. Protein synthesis was measured for in vitro fat bodies from animals treated with combinations of a juvenile hormone analog (JHA = methoprene) and corpora cardiaca (CC) extracts. Ovarian protein content began to increase on Day 5 of adult life and reached its maximum at Days 20-22. Synthesis of proteins secreted by the in vitro fat body increased by 12-fold between emergence and its maximum on Day 18, then declined to nearly its original level by Day 33. Synthesis of nonsecreted, fat body proteins increased by 4-fold between emergence and a maximum on Day 21, then declined. CC extracts and JHA were administered to decapitated females to determine their regulative effects on fat body protein synthesis. The synthesis by the fat body of nonsecreted proteins was increased by both JHA and CC extracts. In contrast, synthesis of secreted proteins increased only in the presence of JHA. CC extracts, alone, had no effects on the synthesis of secreted proteins, but administration of CC extracts in combination with JHA increased the synthesis of the secreted proteins by 55% above that observed with JHA alone. SDS-PAGE of proteins secreted into the medium by the in vitro fat body demonstrated that JHA stimulated the synthesis of specific polypeptides, whereas CC extracts did not affect the synthesis of specific-secreted polypeptides. These results suggest that JH regulates specific protein synthesis by the fat body of B. discoidalis; neurohormones elevate the general capacity of the fat body for protein synthesis and amplify the specific effects of JH.

Age Factors↗

Preliminary characterization and purification of in vitro encapsulation promoting factor: a peptide that mediates insect haemocyte adhesion.

The granular cells and plasmatocytes (PLs) of Heliothis virescens form multicellular aggregations in vitro. This allows capsules to form around suitable targets. Prior trypsinization of the haemocytes abolishes their ability to encapsulate, and this function can be restored by adding plasma to the trypsinized cells. Trypsinized PLs were also unable to spread on a planar glass surface unless normal plasma was present. Plasma was subjected to a variety of treatments to determine the nature of the encapsulation promoting factor (EPF) using the in vitro encapsulation system as a bioassay. The data suggest EPF is a peptide; it is trypsin sensitive and moderately heat stable. Similar results were obtained when using spreading by trypsinized PLs as the bioassay. Dialysis using a 3,500 MW cut-off membrane also abolished encapsulation promoting activity. Protein-free extracts of plasma (crude EPF) has strong activity in both bioassays but does not agglutinate human erythrocytes. A single peak with strong activity in both bioassays was resolved after subjecting crude EPF to reversed-phase HPLC. This active material was purified after additional HPLC with a different solvent system.

Animals↗

Insect hypertrehalosemic hormone: isolation and primary structure from Blaberus discoidalis cockroaches.

A new neurohormone was isolated and structurally characterized that increased hemolymph carbohydrate (trehalose) levels in the cockroach, Blaberus discoidalis. The hormone was isolated in high yield by a rapid HPLC procedure. The sequence, pGlu-Val-Asn-Phe-Ser-Pro-Gly-Trp-Gly-Thr-NH2, was suggested from gas-phase Edman degradation of a peptide fragment of the natural peptide after deblocking with pyroglutamate aminopeptidase. The structure was confirmed by synthesis of the suggested sequence. The synthetic peptide had identical chromatographic and biological properties as the natural peptide.

Amino Acid Sequence↗

Properties of an in vitro bioassay for hypertrehalosemic hormone of Blaberus discoidalis cockroaches.

The ability of the fat body to respond to the hypertrehalosemic action of corpora cardiaca-allata (CC + CA) extracts was measured in vitro with tissue from adult, male Blaberus discoidalis cockroaches. Only trehalose increased in the hemolymph when gland extracts were injected in vivo; likewise, only trehalose levels responded when fat bodies were exposed to gland extracts in vitro. Although fat bodies from 0-day-old animals gave the least response to gland extracts, all animals 5 or more days old showed elevated responses. A linear dose-response occurred between 0.02 and 0.08 CC + CA pair. Fat bodies did not show an in vitro, hypertrehalosemic response to insulin, glucagon, proctolin, dopamine, epinephrine, norepinephrine, serotonin, octopamine, or tyramine. A strong hypertrehalosemic response was found to both synthetic adipokinetic hormone and red-pigment concentrating hormone; however, reversed-phase, thin-layer chromatography of CC + CA extracts and adipokinetic hormone indicated that the hypertrehalosemic effect and the adipokinetic hormone were separate. This in vitro system comprises a relatively rapid, simple, and accurate bioassay for the hypertrehalosemic neurohormone of insects.

Adipose Tissue↗

Isolation of neurosecretory granules containing cytochromogenic hormone activity from the corpora cardiaca of Blaberus discoidalis cockroaches.

Neurosecretory granules (NSG) were isolated by differential centrifugation from a homogenate of the corpora cardiaca-corpora allata complex of newly emerged, adult, female Blaberus discoidalis cockroaches. By centrifuging at 11,000 g, the procedure yielded a final pellet that was enriched in small, membrane-bound vesicles (103 nm mean diameter). This pellet contained 61% of the cytochromogenic activity present in the original homogenate at an 11-fold increase in concentration. A pellet formed at 4000 g contained a mixture of mitochondria and larger NSG (146 nm mean diameter) and most of the hindgut-stimulating hormone activity. This procedure provided a quick, simple method for purifying neurosecretory granules and separating them into two categories of size and endocrine content.

Animals↗

Isolation, identification and synthesis of locustamyotropin (Lom-MT), a novel biologically active insect peptide.

A peptide that stimulates the spontaneous contractions of the hindgut of Leucophaea maderae has been purified from extracts of brain-corpora cardiaca/corpora allata-subesophageal ganglion complexes of 9000 adult Locusta migratoria and was designated locustamyotropin or Lom-MT. The primary structure of this 12 residue peptide has been determined Gly-Ala-Val-Pro-Ala-Ala-Gln-Phe-Ser-Pro-Arg-Leu-NH2. The C-terminal sequence (Phe-Ser-Pro-Arg-Leu-NH2) is identical to the C-terminal pentapeptide of the pheromone biosynthesis activating neuropeptide, recently isolated from Heliothis zea, and is also similar to the C-terminal of leucopyrokinin of Leucophaea. Synthetic Lom-MT showed biological as well as chemical characteristics, indistinguishable from those of native Lom-MT. In locust preparations, Lom-MT provoked an increase in frequency, amplitude and tonus of contractions of the oviduct, but was inactive in the same conditions on the locust hindgut preparation.

Amino Acid Sequence↗

Structure-activity relationships for inhibitory insect myosuppressins: contrast with the stimulatory sulfakinins.

Unusual among insect neuropeptides, the decapeptide myosuppressins are capable of inhibiting contractions of visceral muscle, including the isolated cockroach hindgut. The C-terminal pentapeptide Val-Phe-Leu-Arg-Phe-NH2 has been identified as the myosuppressin active core, the minimum number of residues required to elicit hindgut myoinhibitory activity. Activity of the same magnitude as the parent neuropeptide requires the C-terminal heptapeptide fragment Asp-His-Val-Phe-Leu-Arg-Phe-NH2. Evaluation of a series of substitution analogs delineates structural features critical for myoinhibitory activity within this important fragment. The branched, hydrophobic residues in myosuppressin position 6 (Val) and particularly position 8 (Leu), their absence in the myostimulatory sulfakinins, and the different roles played by the shared Asp residue (myosuppressin position 4; leucosulfakinin position 5) in peptide-receptor interaction, account in large degree for the contrasting biological activities elicited by these otherwise structurally similar peptide families. The results may have broad significance for other invertebrate myotropic systems, such as the locust heart and the pharyngeal retractor muscle of the mollusc Helix aspersa.

Amino Acid Sequence↗

Structure-activity relationships for myotropic activity of the gastrin/cholecystokinin-like insect sulfakinins.

The sulfakinins constitute a family of real and putative peptide sequences characterized from the cockroach Leucophaea maderae (leucosulfakinin subfamily) and fruitfly Drosophila melanogaster (drosulfakinin subfamily) with homology to the sulfated mammalian hormones gastrin II and cholecystokinin (CCK). The leucosulfakinin (LSK) subfamily of neuropeptides stimulate contractions of the isolated cockroach hindgut. In this paper, we have ascertained some of the primary structural requirements of the sulfakinins for myotropic (muscle-contracting) activity. The myotropic "active core" of this family has been determined to be the C-terminal hexapeptide, though the C-terminal octapeptide (Glu-Asp-Tyr(SO3H)-Gly-His-Met-Arg-Phe-NH2) is required for full activity. The LSKs demonstrate considerable tolerance to Ala substitution in positions 7 and 9 within the active core without complete loss of activity. Conversely, Ala substitution in positions 8, 10 and 11 led to inactive compounds. Basicity is a critical feature of LSK position 10, while aromatic character is an important characteristic for positions 8 and 11 for myotropic activity. Only trace activity could be observed upon replacement of the Tyr(SO3H) residue in LSK-position 6 with a Ser(SO3H). One analog ([3MeHis8] LSK) proved more active as a contractile stimulant than the natural product, while another ([7-11,Tyr(SO3H)7] LSK), conversely, demonstrated inhibition of spontaneous contractions of the cockroach hindgut.

Amino Acid Sequence↗