PubMed Health⌕ Search

Biomedical subjects

B G Jenks

Publications and source records attributed to B G Jenks.

At least 109 records · Page 6Linked to original sources
PubMed1986

The pituitary adrenocorticotropes originate from neural ridge tissue in Xenopus laevis.

A series of grafting experiments was conducted to determine pituitary origins prior to brain tube closure in Xenopus laevis. Extirpation experiments indicated that the ventral neural ridge (VNR) tissue of stage-18+ embryos was essential for pituitary development. Bolton-Hunter reagent was used to label stage-18+ VNR tissue with 125I, and this tissue was then returned to the donor and its subsequent ontogenesis followed. Labelled tissue was ultimately found in the ventral hypothalamus, the ventral retina, and the anterior pituitary. Using immunocytochemical techniques with antisera to adrenocorticotropin (ACTH), it was found that some of the VNR-derived cells were corticotropes. A region of the nucleus infundibularis which was radioactive labelled also gave ACTH-positive immunoreaction. This might indicate that some ACTH-containing neurones of the hypothalamus are VNR in origin. We suggest that stage-18+ VNR is the site of attachment of brain and anterior pituitary ectoderm. Part of this adherence point is eventually incorporated into the anterior pituitary and will form corticotropes. It is concluded that the ventral retina, the preoptic region of the hypothalamus, some hypothalamic ACTH-immunoreactive cells, and the most anterior portion of the adenohypophysis are all ventral neural ridge in origin.

Adrenocorticotropic Hormone↗
PubMed1985

The functional significance of glycosylation of pro-opiomelanocortin in melanotrophs of the mouse pituitary gland.

Pro-opiomelanocortin (POMC) is a glycoprotein precursor for a number of neuropeptides and peptide hormones. The functional significance of the glycosylation of POMC has never been established. Using the antibiotic tunicamycin to block glycosylation of the prohormone in the mouse pars intermedia, we have compared processing of non-glycosylated prohormone with that of glycosylated prohormone in pulse-chase experiments. The peptides produced from non-glycosylated prohormone were shown to be correct cleavage products. Therefore it was concluded that, with the possible exception of peptides from the N-terminal region of the prohormone, the carbohydrate on POMC plays no role in directing cleavage or in protecting the prohormone from random proteolysis. Tunicamycin treatment retarded N-terminal acetylation of melanotrophin but had no apparent effect on acetylation of beta-endorphin. The mouse pars intermedia synthesizes two forms of POMC which differ in their degree of glycosylation. Our results indicated that, during secretion, the melanotrophs make no distinction between peptides derived from the two prohormones.

Adrenocorticotropic Hormone↗
PubMed1985

[Intermediate lobe of the amphibian pituitary gland: an endocrine gland with multiple secretions and under multi-hormonal control].

The cells of the frog pars intermedia synthesize a 36 000 (36K) protein called proopiomelanocortin (POMC). After [3H]glucosamine incorporation, separation of newly synthesized products by SDS-polyacrylamide gel electrophoresis showed that this 36K protein was glycosylated. Tryptic mapping revealed only one site of glycosylation and showed that the carbohydrate side-chain was located in the N-terminal region of POMC. The 36K protein was not released by the melanotrophs, but it generated, through specific intracellular proteolytic cleavage, a number of smaller peptides which were subsequently released. These peptides were identified by various methods including selective amino-acid incorporation, HPLC purification, acid-urea gel electrophoresis, tryptic and chymotryptic mapping, assay of melanotropic activity, radioimmunoassays and immunoprecipitations. Some of the newly synthesized N-terminal (18K) fragment of the POMC was secreted intact while a portion of it was further processed, via an intermediate peptide, to give mature gamma-MSH. All three of these peptides were glycosylated. In addition, the mature peptide (gamma-MSH) exhibited a low but significant melanotropic activity. The C-terminal portion of the prohormone was very rapidly processed to give des N alpha-acetyl alpha-MSH, corticotropin-like-intermediate lobe peptide (CLIP) and beta-endorphin. Authentic alpha-MSH was always absent in cellular extracts: acetylation to give rise to alpha-MSH was a late enzymatic process strictly linked to hormonal release. Since acetylation of alpha-MSH is required for full biological activity of this peptide, it is possible to conceive that this later step could be under neuroendocrine control. Using the perifusion technique we have been able to show the complexity of the control mechanisms regulating amphibian melanotrophs. It is generally accepted that the aminergic innervation of the intermediate lobe of the pituitary is involved in the hypothalamic control of melanotropin release. We have demonstrated that, in amphibians, dopamine inhibits alpha-MSH secretion through D2-type dopaminergic receptors whereas norepinephrine and (or) epinephrine stimulate alpha-MSH secretion via beta-adrenergic receptors. The existence of peptidergic fibers within parenchymal cells of the pars intermedia has been demonstrated. Evidence for TRH-containing fibers has been obtained by immunohistochemistry. Using a specific radioimmunoassay for TRH, we have confirmed the presence of TRH in the neurointermediate lobe of the frog. We have shown that TRH is a powerful MSH-releasing factor in these animals.(ABSTRACT TRUNCATED AT 400 WORDS)

Amphibians↗
PubMed1984

The development of the pars intermedia and its role in the regulation of dermal melanophores in the larvae of the amphibian Xenopus laevis.

The ontogenesis of biosynthesis of pro-opiomelanocortin (POMC)-related peptides in the pars intermedia of Xenopus laevis tadpoles was studied. The results were related to the capacity of the animal to adapt to background color through regulation of pigment dispersion in dermal melanophores. Using immunocytochemical techniques with antisera to alpha-melanophore-stimulating hormone (alpha-MSH), it was revealed that this peptide first appeared at developmental stage 37/38, just prior to the animal's ability to adapt to background. It was shown that pigment dispersion in melanophores between stages 33 and 39 was not dependent on melanotropins of pituitary origin. Using in vitro biosynthetic studies it was possible to follow POMC biosynthetic activity, its processing and the release of peptides from stage 48 onward. Among the newly synthesized peptides observed were a gamma 3-MSH-like peptide, des-N-alpha-acetyl-alpha-MSH, alpha-MSH, and two endorphin-like peptides. By stage 57 a biosynthetic pattern almost identical to that of the adult pars intermedia had evolved. It was concluded that stage 39/40 is a critical stage in the simultaneous development of a number of the components involved in the neuroendocrine control of background adaptation.

Adaptation, Physiological↗
PubMed1983

Hydroxylamine cleavage of proteins in polyacrylamide gels.

A modification of the hydroxylamine cleavage of proteins is presented in which proteins were cleaved while immobilized in the matrix of a polyacrylamide gel. The reaction under these conditions retains its high specificity for Asn-Gly bonds and has the advantage that the gel matrix, acting as a carrier, facilitates simultaneous treatment of many samples, and contributes to a high recovery efficiency (60-90%) of the cleavage products. The cleavage is performed with individual protein bands excised from dried slab gels after detection by staining, autoradiography, or fluorography. The procedure can be easily combined with other techniques to further characterize the cleavage fragments. Also a two-dimensional version of the cleavage method was developed, which allows rapid recognition of interrelationships between proteins in a complicated mixture. The versatility of the procedure is demonstrated in a number of applications. Highly related strains of murine leukemia viruses were easily distinguished from one another by the unique cleavage patterns of their gag- and env-precursor polypeptides. Comparing the env-precursor gPr82env synthesized in the presence or absence of tunicamycin with its cell-free synthesized counterpart, revealed the presence of an amino-terminal signal sequence. Cleavage patterns of pro-opiomelanocortin (POMC) from three different species revealed a high degree of homology between rat and mouse POMC, whereas Xenopus POMC was very different. Regions to which carbohydrates are attached could be identified by comparing glycosylated and unglycosylated forms of POMC. Combining the hydroxylamine cleavage procedure with immunological characterization of the fragments showed a small but significant difference between the amino-terminal sequences of the recombinant transforming protein P120 of Abelson murine leukemia virus and of its parent molecule Pr65gag of Moloney murine leukemia virus.

Animals↗
PubMed1983

In vivo biosynthesis of melanotropins and related peptides in the pars intermedia of Xenopus laevis.

To study in vivo biosynthesis of pars intermedia peptides in Xenopus laevis, [3H]lysine was administered by an osmotic minipump via a cannula inserted near the pituitary gland. Following extraction of the neurointermediate lobe, high-performance liquid chromatography was used to separate the newly synthesized peptides. In black-background adapted animals, [3H]lysine was incorporated into a number of peptides. The elution characteristics of these peptides corresponded exactly with those of peptides synthesized during in vitro incubation of neurointermediate lobes, and which were identified as des-N alpha-acetyl-alpha-MSH, a gamma-MSH-like peptide, two corticotropin-like intermediate lobe peptides, and two forms of endorphin. In white-background adapted Xenopus, practically no synthesis of pars intermedia peptides occurred. Transfer of black-adapted toads to a white background at the beginning of infusion led to storage of newly synthesized peptides. When such animals were maintained on a white background for 10 days, des-N alpha-acetyl-alpha-MSH, but not alpha-MSH, was present in the pars intermedia; this supports the notion that des-N alpha-acetyl-alpha-MSH constitutes the "storage form" of alpha-MSH.

Adaptation, Physiological↗
PubMed1983

The frog pars intermedia contains only the non-acetylated form of alpha-MSH: acetylation to generate alpha-MSH occurs during the release process.

Pulse-chase experiments revealed that the frog pars intermedia synthesizes the desacetyl form of alpha-MSH. Its structure was shown to be similar, if not identical, to the mammalian structure. During release two additional peptides derived from desacetyl alpha-MSH appeared, one being alpha-MSH. We conclude that the N-acetylation of newly synthesized MSH is associated with release of the hormone. Radioimmunoassays and bioassays showed that the non-acetylated peptide is the only tissue form of MSH and confirmed that acetylation is linked to release.

Acetylation↗
PubMed1983

Biosynthesis of pro-opiomelanocortin-derived peptides in the mouse neurointermediate lobe.

This report concerns biosynthetic studies conducted with neurointermediate lobes of the mouse pituitary gland. High performance liquid chromatography was used to resolve newly synthesized peptides after in-vitro incubation of lobes with radioactive amino acids. Among the newly synthesized peptides identified were alpha-MSH, des-N alpha-acetyl-alpha-MSH, two forms of corticotrophin-like intermediate lobe peptide and beta-endorphin. The biosynthesis of a glycosylated gamma 3-MSH-like peptide was also demonstrated. While no newly synthesized beta-MSH could be identified, a peptide designated gamma-lipotrophin was found. Pulse-chase analysis revealed that the major biosynthetic pathway leading to the production of alpha-MSH involved the acetylation of the des-acetyl form of this peptide. Furthermore, it was evident that newly synthesized beta-endorphin was largely converted to modified forms of this peptide; most of the terminal product was probably N-acetylated endorphin.

Adrenocorticotropic Hormone↗
PubMed1983

Biosynthesis of pro-opiomelanocortin-related peptides in the neurointermediate lobe of the pituitary gland of the rainbow trout (Salmo gairdneri).

This study concerns the biosynthesis of a number of peptides in the neurointermediate lobe of the rainbow trout, Salmo gairdneri. Among the newly synthesized peptides positively identified were four forms of alpha-MSH, a beta-MSH and two forms of endorphin. The identification of two of the products as sequentially distinct endorphins suggests that the trout neurointermediate lobe synthesizes two pro-opiomelanocortin (POMC) molecules. This may account for the large number of peptides synthesized and secreted by the trout neurointermediate lobe. The identification of some of the products is discussed. Pulse-chase analysis revealed no positive identification of a precursor molecule, which may indicate that trout POMC is very rapidly processed into smaller products. All of the newly synthesized peptides were secreted concomitantly. There was little if any evidence of post-translational modification of any of the peptides during their secretion.

Amino Acids↗
PubMed1982

Biosynthesis of pairs of peptides related to melanotropin, corticotropin and endorphin in the pars intermedia of the amphibian pituitary gland.

This study concerns the biosynthesis of a number of peptides in the neurointermediate lobe of the pituitary gland of the aquatic toad, Xenopus laevis. Using pulse-chase incubations in vitro and high-performance liquid chromatographic analysis, it could be shown that these peptides are synthesized through processing of a prohormone, pro-opiomelanocortin; all peptides were released into the incubation medium. On the basis of electrophoretic analysis, selective amino acid incorporation and immunoprecipitation, as well as peptide mapping by high-performance liquid chromatography, the peptides were classified into three distinct groups: two related to melanocyte-stimulating hormone (melanotropin), two related to adrenocorticotropic hormone (corticotropin) and two endorphin-like peptides. Using tryptic and chymotryptic maps of synthetic alpha-melanotropin and des-Ac alpha N-alpha-melanotropin as references, one of the melanotropin-like peptides was identified as des-Ac alpha N-alpha-melanotropin; the other one represents neither alpha-melanotropin nor any other known melanotropic peptide. The two peptides that were immunologically related to corticotropin had characteristics consistent with a structures resembling a peptide previously named 'corticotropin-like intermediate lobe peptide', corticotropin-(18-39). The two endorphin-like peptides, although highly related, do not have the same primary structure. In view of the apparent structural differences between the two peptides in each group, the possible occurrence of two prohormones is discussed.

Adrenocorticotropic Hormone↗
PubMed1982

Biosynthesis of two structurally different pro-opiomelanocortins in the pars intermedia of the amphibian pituitary gland.

This study reports the biosynthesis of two forms of pro-opiomelanocortin in the pars intermedia of the pituitary gland of the African clawed toad Xenopus laevis. The two forms could be resolved by dodecyl sulphate gel electrophoresis on a 9-16% acrylamide gradient and their molecular weights were 38200 and 37300. Incubation of neurointermediate lobes with [3H]glucosamine followed by tryptic digestion of the newly synthesized glycoproteins, revealed that both prohormones have only one glycosylated site, namely within the region corresponding to gamma 3-melanotropin. Biosynthesis of proteins in lobes treated with tunicamycin to prevent glycosylation again resulted in the production of two pro-opiomelanocortins (Mr 35000 and 34200), indicating that the two forms differ in their primary structure. This notion was corroborated by the results of tryptic mapping of the newly synthesized prohormones. The maps showed that the primary structures of the two forms of pro-opiomelanocortin differ in at least two parts of the molecules, one part concerning the endorphin region.

Animals↗
PubMed1982

Biosynthesis of a gamma 3-melanotropin-like peptide in the pars intermedia of the amphibian pituitary gland.

This study reveals the biosynthesis of a gamma 3-melanotropin-like peptide in the pars intermedia of the pituitary gland of the aquatic toad Xenopus laevis. Pulse-chase experiments in vitro showed that this product is synthesized through processing of a prohormone, pro-opiomelanocortin, and that it is released into the incubation medium. The peptide immunoprecipitated with antiserum to gamma 3-melanotropin, appeared to be a glycopeptide and displayed melanotropic activity. This last observation together with the results of tryptic and chymotryptic peptide mapping of the newly synthesized product indicates that Xenopus gamma 3-melanotropin is structurally different from the proposed mammalian gamma-melanotropins.

Animals↗
PubMed1979

Biosynthesis of MSH and related peptides in the pars intermedia of the mouse: a pulse-chase analysis.

Protein biosynthesis in neurointermediate lobes of mouse pituitaries was investigated using pulse and pulse-chase techniques with [3H]lysine. Electrophoretic analysis of lobe homogenates on acid-urea gels resolved 11 labeled products. One was a large protein which was rapidly synthesized during pulse-incubations and disappeared during chase incubations. Three of the products increased during chase incubations, suggesting a precursor-product mode of biosynthesis for these chasde peptides. One of these three products co-migrated with synthetic alpha-MSH and also corresponds to the major peak of mouse neurointermediate lobe MSH bioactivity and immunoactivity on electrophoretograms. Another case of these peptides has electrophoretic properties similar to those of ACTH.

Animals↗
PubMedDate not supplied

Age-related changes in adenohypophyses of mice with the hereditary obese hyperglycemic syndrome (ob/ob) in relation to the diabetic state of the animals.

Mice with hereditary, monogenic obese-hyperglycemic syndrome (ob/ob) were shown to have smaller adenohypophyses in comparison to their normal littermates from an age of 4 weeks onward. The size of the pars intermedia of obese mice was not, however, significantly different from that of normal littermates. Morphometric criteria strongly suggest hyperactivity of the cells of the pars intermedia of obese hyperglycemic mice. The enhanced secretion of bioactive peptides of the pars intermedia may contribute to the diabetic aspects of the obese-hyperglycemic syndrome. The hyperactivity of the pars intermedia is likely to be allophenic.

Aging↗