Existence of a common precursor to ACTH and endorphin in the anterior and intermediate lobes of the rat pituaitary

Eipper, Betty A.; Mains, Richard E.

doi:10.1002/jss.400080304

Cited by 211 publications

(75 citation statements)

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“…Comparison of the ACTH/f3-endorphin precursors and processed products synthesized by the toad intact intermediate lobe with those synthesized by the AtT-20 mouse anterior pituitary cell line and rat pituitary cell suspensions (27) The significance of having two common precursors is unclear. Possibly, different sets of processed peptides are cleaved from each of the precursors [e.g., the glycosylated ACTH forms may be derived from the more glycosylated precursor (peak "a1"), and the unglycosylated forms, from the other].…”

Section: Resultsmentioning

confidence: 99%

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Immunological evidence for two common precursors to corticotropins, endorphins, and melanotropin in the neurointermediate lobe of the toad pituitary.

Loh¹

1979

Proc. Natl. Acad. Sci. U.S.A.

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The biosynthesis of corticotropin (ACTH1-39), and a-melanotropin in the toad intermediate lobe was studied by using immunoprecipitation procedures with antisera specific for these peptides. Intermediate lobes were pulse-incubated with [3H phenylalanine and then chase-incubated for varying periods; the radioactive proteins were immunoprecipitated. Immunoprecipitates were separated by acidic urea or sodium dodecyl sulfate polyacrylamide gel electrophoresis. Evidence from the pulse-chase and sequential immunoprecipitation studies using antisera to ACTH and 3-endorphin suggests that the toad intermediate lobe synthesizes two common precursors (apparent M, 32,000 and 29,500) containing both the ACTH and 3-endorphin sequences. from mouse appear to synthesize not only ACTH but also ,B-LPH from cleavage of a common precursor of approximately 31,000 daltons. In addition, a polyadenylylated RNA fraction from such tumor cells has been translated in the reticulocyte and wheat germ cell-free systems and, among the translation products, is a 28,500-dalton protein that is precipitated by anti-ACTH antisera and includes tryptic digestion sequences that are indistinguishable from those of ACTH and ,15 After the pulse, the neurointermediate lobes were homogenized. A group of 0.5-hr pulsed lobes were subjected to a chase in amphibian Ringer's solution containing unlabeled phenylalanine (1 mM) for 1 or 3 hr before homogenization. The medium after a 3-hr chase was also collected for analysis of released peptides. Lobes were homogenized in 5 M acetic acid containing 5 mg of bovine serum albumin per ml and 1% phenylmethylsulfonyl fluoride (30 mg/ml)/iodoacetamide (30 mg/ml) in absolute ethanol, freshly made within hours of its use. Samples were frozen and thawed three times on dry ice and extracted at 4°C for 12-16 hr. Extracts were then lyophilized.Abbreviations: LPH, lipotropin; ACTH, corticotropin; MSH, melanotropin; NaDodSO4, sodium dodecyl sulfate. 796The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U. S. C. §1734 solely to indicate this fact.

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Section: Resultsmentioning

confidence: 99%

“…Since this manuscript was submitted, it has been shown that cell suspensions from anterior and neurointermediate lobes of the rat pituitary have a biosynthetic pathway for ACTH, R-LPH, and f3-endorphin similar to that of the toad neurointermediate lobe (27).…”

Section: Resultsmentioning

confidence: 99%

Immunological evidence for two common precursors to corticotropins, endorphins, and melanotropin in the neurointermediate lobe of the toad pituitary.

Loh¹

1979

Proc. Natl. Acad. Sci. U.S.A.

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“…1) Three immunoreactive forms of ACTH are characterized by their apparent molecular weight (MW) (Orth et al, 1970(Orth et al, , 1973 (Eipper and Mains, 1978 ;Mains and Eipper, 1979) which also contains the amino acid sequences of both the /3LPH and 16-K fragments located near the N-end of the precursor (Lowry et al, 1976 ;Eipper and Mains, 1978 ;Nakanishi et al, 1979 Dupouy, 1980b (Dupouy, 1976 in the pituitary gland of the sheep fetus and the lamb which contains abundant material of high molecular weight (Silman, 1979 ;Silman et al, 1979 ;Roebuck et al, 1980 (Silman et al, 1978).…”

Section: Discussionmentioning

confidence: 99%

Heterogeneity of adrenocorticotrophin in the pituitary gland and plasma during the perinatal period

Dupouy¹,

Chatelain²

1985

Reprod. Nutr. Dévelop.

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“…We used rabbit polyclonal antisera to RESP18 (JH1162 and JH1163, 1:4000 dilution), POMC (Georgie, 1:2000), AVP and OXY (V4 and V10, 1:1000 dilution; gift of Dr. Hal (Eipper and Mains 1978;Bloomquist et al 1994a,b;Darlington et al 1996a;Paquet et al 1996). We also used monoclonal antisera to synaptophysin (#902 314, 1:1500; Boehringer Mannheim, Indianapolis, IN) and synaptotagmin (1:3000 dilution; gift of Dr. Richard Scheller).…”

Section: Antiseramentioning

confidence: 99%

Expression of RESP18 in Peptidergic and Catecholaminergic Neurons

Darlington

Schiller

Mains

et al. 1997

J Histochem Cytochem.

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SUMMARYWe examined the expression of regulated endocrine-specific protein of 18-kD (RESP18) in selected peptidergic and catecholaminergic neurons of adult rat brain. In the hypothalamic paraventricular, supraoptic, and accessory nuclei, RESP18 mRNA was highly expressed in neurons immunostained for oxytocin and vasopressin. RESP18 mRNA was also highly expressed in paraventricular nucleus neurons immunostained for corticotropinreleasing hormone, thyrotropin-releasing hormone, and somatostatin. RESP18 mRNA was expressed in POMC cells of the arcuate nucleus, in neuropeptide Y cells of the dorsal tegmental nucleus, lateral reticular nucleus, and hippocampus, and in brainstem catecholaminergic neurons. RESP18 mRNA expression was high in all paraventricular and arcuate neurons, but RESP18 protein was detectable in the perikarya of a subset of these neurons, suggesting an important post-transcriptional component to the regulation of RESP18 expression. RESP18 antisera immunostained perikarya but not axon fibers or terminals. Subcellular fractionation of homogenates of several hypothalamic nuclei identified RESP18 protein in fractions enriched in endoplasmic reticulum. The presence of 22-and 24-kD RESP18 isoforms in the neural lobe of the pituitary indicated that some RESP18 protein exited the endoplasmic reticulum. The post-transcriptional regulation of RESP18 expression and localization of RESP18 protein primarily to the endoplasmic reticulum suggests that RESP18 plays a regulatory role in peptidergic neurons. R egulated endocrine-specific protein 18 (RESP18) is a novel 18-kD protein with limited homology to a short region of several receptor protein tyrosine phosphatases (Bloomquist et al. 1994a;Schiller et al. 1995;Darlington et al. 1996a). The RESP18 cDNA encodes a functional NH 2 terminal signal sequence and newly synthesized RESP18 enters the lumen of the secretory pathway (Bloomquist et al. 1994a;Schiller et al. 1995). RESP18 is expressed in AtT-20 corticotrope tumor cells and immunostaining and subcellular fractionation analyses identified the protein in the endoplasmic reticulum (Schiller et al. 1995). The RESP18 protein has a half-life of less than 15 min (Bloomquist et al. 1994b) but can be stabilized by calpain protease inhibitors (Schiller et al. 1995). Although RESP18 is not normally secreted by AtT-20 cells, RESP18 is found in the media of AtT-20 cells treated with calpain inhibitors or overexpressing exogenous RESP18 (Schiller et al. 1995). Under these conditions, a 21-kD isoform of RESP18 is observed in the media. Higher molecular mass RESP18 isoforms are also found in vivo and may result from O-glycosylation (Schiller et al. 1995;Darlington et al. 1996b;Schiller and Darlington 1996).RESP18 expression is limited to brain, pituitary, peripheral endocrine tissues, and sperm, suggesting that its function is specific to neurons, endocrine cells, and sperm (Bloomquist et al. 1994a,b; Darlington et al. 1996a,b;Schiller and Darlington 1996). In rat pituitary, RESP18 mRNA has been identified in corticotropes, t...

show abstract

Existence of a common precursor to ACTH and endorphin in the anterior and intermediate lobes of the rat pituaitary

Cited by 211 publications

References 35 publications

Immunological evidence for two common precursors to corticotropins, endorphins, and melanotropin in the neurointermediate lobe of the toad pituitary.

Immunological evidence for two common precursors to corticotropins, endorphins, and melanotropin in the neurointermediate lobe of the toad pituitary.

Heterogeneity of adrenocorticotrophin in the pituitary gland and plasma during the perinatal period

Expression of RESP18 in Peptidergic and Catecholaminergic Neurons

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