Molecular forms of α-melanocyte-stimulating hormone in the canine pituitary anterior and intermediate lobe

Reverse-phase high pressure liquid chromatography (HPLC) and radioimmunoassay (RIA) were used to determine the distribution of naturally occurring forms of α-melanocyte-stimulating hormone (α-MSH) in acid extracts of pars intermedia (PI) and anterior lobe (AL) tissue from canine and rat pituitary. Similarly, intracellular and secreted forms of α-MSH were determined using cultured canine PI and AL cells. Rat PI tissue contained predominantly diacetyl-α-MSH, while monoacetyl-α-MSH was the most abundant form in canine PI. In both canine and rat AL tissue extracts desacetyl-α-MSH was the major form of α-MSH. The profile of α-MSH contained in and secreted into culture medium by canine PI cells was found to be very similar to that in PI tissue extracts. The proportion of monoacetyl-α-MSH and diacetyl-α-MSH secreted by cultured canine AL cells and contained in extracts of AL cells in culture, however, was much higher than that in tissue extracts. These results indicate that in the dog, as in all other mammalian species studied, acetylated forms of α-MSH predominate in PI tissue, while nonacetylated α-MSH is the major form in AL tissue. It appears, however, that acetylation of α-MSH may occur in cultured canine AL cells, possibly as a result of the absence of factors that normally inhibit acetyltransferase in vivo or as a consequence of culture conditions.     

Presence of alpha-melanocyte-stimulating hormone in bovine pituitary intraglandular colloid of intermediate lobe origin

Alpha-melanocyte-stimulating hormone (alpha-MSH), one of several peptide hormones originating in the intermediate lobe of the pituitary as proopiomelanocortin, was discovered in bovine pituitary intraglandular colloid by using radioimmunoassay. The quantity of alpha-MSH varied from 5 to 368 micrograms/mg protein in the three pools. The importance of this finding is discussed in light of the possibility that the colloid is a transport medium for alpha-MSH and other intermediate lobe hormones.     

Adenosine 3',5'-cyclic monophosphate stimulates secretion of alpha-melanocyte-stimulating hormone from permeabilized cells of the intermediate lobe of the rat pituitary gland

The effect of Mg-ATP and cyclic AMP on the secretion of alpha-melanocyto-stimulating hormone (alpha-MSH) from electrically permeabilized cells of rat intermediate lobe (IL) were investigated. Addition of exogenous Ca2+ stimulated alpha-MSH secretion in a concentration- (EC50 = 4.8 microM) and temperature-dependent manner. This Ca2+-evoked secretion was further enhanced by Mg-ATP and cyclic AMP. Mg-ATP was required for the fully secretory response in the electrically permeabilized IL cells and the maximal secretion was reached at 1 mM. Cyclic AMP in the presence of GTP gamma S also potentiated Ca2+-evoked alpha-MSH secretion to the same magnitude as Mg-ATP. In the absence of Ca2+ both the cyclic AMP and Mg-ATP did not stimulate alpha-MSH secretion from IL cells. The data suggest that Mg-ATP and cyclic AMP may modulate directly the secretory components rather than change intracellular concentration of free Ca2+.     

Rat atrial natriuretic factor suppresses proopiomelanocortin-derived peptides secretion from both anterior and intermediate lobe cells and growth hormone release from anterior lobe cells of rat pituitary in vitro

Synthetic rat atrial natriuretic factor (ANF) was found to attenuate, in a dose-dependent manner, basal and corticotropin-releasing factor-induced secretion of proopiomelanocortin-derived peptides from cultured anterior and intermediate lobe cells of rat pituitary. ANF was also found to suppress basal and growth hormone-releasing factor-stimulated secretion of growth hormone from anterior lobe cells of rat pituitary. These results, together with reports of the existence of ANF-positive neurons in the hypothalamus and ANF-positive fibers in the median eminence, suggest that hypothalamic ANF is probably involved in the regulation of pituitary hormone secretion, especially that of proopiomelanocortin-derived peptides and growth hormone.     

Serotonin immunoreactivity in the intermediate lobe of the rat pituitary.

Immunocytochemical staining for serotonin (5-HT) in paraffin-embedded sections of rat pituitary resulted in the localization of reactive nerve fibres and cell bodies in the intermediate lobe. Immunostaining was also found in the anterior and posterior lobes. Labelled nerve fibres appear to enter the intermediate lobe from the neural lobe through the interlobular spaces. These fibres are relatively scarce and lightly stained. Neuroglandular contacts were identified between varicose nerve endings containing serotonin and immunoreactive perykarion. It is not clear whether intermediate lobe cells produced 5-HT themselves or, alternatively, these cells take in 5-HT from serotoninergic nerve terminals.     

Immunoreactive calcitonin in the intermediate lobe of the pituitary gland.

Immunoreactive-calcitonin was observed in all cells of the intermediate lobe of the rat pituitary gland. It may thus be a fragment of the ≥31, 000 daltons precursor molecule of adrenocorticotropin and β-lipotropin.     

Melanin concentrating hormone. V. Isolation and characterization of alpha-melanocyte-stimulating hormone from frog pituitary glands

The structure of alpha-melanocyte-stimulating hormone (alpha-MSH) has been determined in the pars intermedia of the frog Rana ridibunda. Pulse-chase labeling of frog neurointermediate lobes with selective amino acids revealed that the composition of frog alpha-MSH is similar to that of alpha-MSH from all mammalian species yet studied. Tryptic mapping of nexly synthetized alpha-MSH generated two fragments with the following amino acid composition: (T1) Trp, Pro, Lys, Gly, Val and (T2) Tyr, Arg, Phe, His, Ser, Glu. Concurrently, alpha-MSH was purified from 100 neurointermediate lobes to apparent homogeneity by reverse-phase HPLC. The sequence of the peptide determined by automated Edman degradation was Ser-Tyr-Ser-Met-Glu-His-Phe-Arg-Trp-Gly-Lys-Pro-Val. The structure of frog alpha-MSH is thus identical to mammalian des-N alpha-acetyl alpha-MSH and differs from the sequence of toad (Xenopus laevis) alpha-MSH only by the first residue (Ser instead of Ala). These results confirm that the sequence of alpha-MSH has been highly preserved during evolution.     

Endorphin activity in anterior, intermediate, and posterior pituitary.

The content of endogenous morphine-like substance in bovine pituitary and brain was determined by an opiate radioreceptor assay. The intermediate lobe was most concentrated in activity and the brain least concentrated. Most of the endorphin is obtained in a 120 000 g-min fraction from pituitary or brain homogenates.     

Beta-endorphin in pituitary intraglandular colloid of intermediate lobe origin

Radioimmunoassay (RIA) detected the presence of beta-endorphin in the intraglandular colloid (IGC) of bovine pituitary intermediate lobe origin. The amount of beta-endorphin recovered in each of twelve samples ranged from 0.15 to 218.30 pmol/mg protein. A second group of assays [amino acid analysis, high performance liquid chromatography (HPLC) and mass spectral analysis] confirmed the RIA findings in another series of colloid samples. Approximately 75 pmol was collected from eight pooled glands. beta-Endorphin is an addition to the list of peptide hormones (e.g., methionine-enkephalin, adrenocorticotropin, arginine-vasopressin, alpha-melanocyte-stimulating hormone, beta-lipotropin and somatostatin) previously discovered in IGC by this laboratory.     

Alpha-melanocyte-stimulating hormone secretion from permeabilized intermediate lobe cells of rat pituitary gland. The role of guanine nucleotides

The non-hydrolyzable GTP analogue, guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S) and cyclic AMP potentiated the Ca2+-evoked secretion of alpha-melanocyte-stimulating hormone (alpha-MSH) from permeabilized neurointermediate lobe (IL) cells of rat pituitary gland. The enhancement by Mg-GTP gamma S (100 microM) and cyclic AMP (1 microM) depended on the intracellular Ca2+ concentration (EC50 = 4.8 +/- 1.8 and 4.6 +/- 1.7 microM; mean +/- SE, with and without Mg-GTP gamma S and cyclic AMP, respectively). A similar effect was observed with guanine nucleotide triphosphate (GTP and GppNHp). Mg was absolutely required for this event. Neither Mg-GTP gamma S nor cyclic AMP alone was effective in potentiating alpha-MSH secretion. GDP beta S blocked the Mg-GTP gamma S (100 microM) and cyclic AMP augmented secretion of alpha-MSH. Neither neomycin (which affects the process of inositol 1,4,5-triphosphate-mediated Ca2+ mobilization) or colchicine (which influences microtubule assembly) had an effect on the cyclic AMP and Mg-GTP gamma S potentiation of alpha-MSH secretion. These data suggest that the GTP-binding protein may be involved in the regulation of alpha-MSH secretion after Ca2+ entry into the cells, since the intracellular environment is controlled in the permeabilized cells.     

Adrenocorticotropin production by the intermediate lobe of the rat pituitary

Summary Tissue from the four chambers of the heart of the plaice (Pleuronectes platessa, L.) has been examined in the electron microscope in order to describe the morphology of the heart at a fine structural level.The sinus venosus is a thin walled chamber between 60–90 thick consisting of a connective tissue matrix in which are situated the plexus of the parasympathetic cardiac ganglion and localised bundles of myocardial cells. The myocardial cells do not form a continuous layer but are associated in particular with the region of the cardiac ganglion and are innervated by it.The sino-auricular junction has hitherto been described as a pacemaker region but the myocardial cells in this region are identical in morphology to myocardial cells in other parts of the heart. There is a large complex of nerves, derived from the cardiac plexus, that runs around the junction before branching to innervate the auricle.The myocardial tissues consist of an outer layer of myocardium forming the wall of the heart and a profusion of trabeculae. The endocardium invaginates into the endocardium to divide up the cells into populations of approximately 25 cells in profile. There is no well-defined coronary blood supply although capillaries are occasionally seen. The myocardial cells themselves are small in diameter (3.5–5.5 ) and show some primitive features which are: a short sarcomere (1.4–2.0 ), the absence of any sarcoplasmic reticulum, and very scarce fasciae occludentes. In the atrium in particular, there are many groups of 1500 Å membrane-bound, dense-cored vesicles in the myocardial cells. Ventricular cells contain more myofilaments and mitochondria than do atrial cells and have many vesicles of 0.1–0.3 diameter whose function and contents are unknown.Connective tissue is very evident in the plaice heart, being an integral part of the sinus venosus and the auriculo-ventricular junction and being the sole constituent of the auriculoventricular valve and the bulbus arteriosus.This investigation was carried out during the tenure of an S. R. C. studentshyp awarded to R. M. S.     

The dopaminergic innervation of the intermediate lobe and of the neural lobe of the pituitary gland

The intermediate and the neural lobe of the pituitary gland are innervated by two, virtually independent, groups of dopaminergic neurons which, until recently, were considered as a uniform system and referred to as the "tubero-hypophyseal dopamine system". Some aspects of the separate physiological functions of these neurons in the intermediate and in the neural lobe, of their microanatomy and biochemistry as well as of dopamine release from their terminals are discussed in this review.