Effects of f3-Lipotropin and ,-Lipotropin-derived Peptideson Aldosterone Production in the Rat Adrenal Gland
HIROAKI MATSUOKA,PATRICK J. MULROW,and ROBERTOFRANCO-SAENZ,Department of Medicine, Medical College of Ohio, Toledo, Ohio 43699
CHOHHAOLi, Hormone Research Laboratory, University of California,San Francisco, California 94143
A B S T RA C T To investigate the role of non-ACTHpituitary peptides on steroidogenesis, we studied theeffects of synthetic 3-lipotropin, (8-melanotropin, andp-endorphin on aldosterone and corticosterone stimu-lation using rat adrenal collagenase-dispersed capsularand decapsular cells. f8-lipotropin induced a significantaldosterone stimulation in a dose-dependent fashion(10 nM-1 ,uM). 8-endorphin, which is the carboxy-terminal fragment of 8-lipotropin, did not stimulatealdosterone production at the doses used (3 nM-6 ,uM)./3-melanotropin, which is the middle fragment of 8-lipotropin, showed comparable effects on aldosteronestimulation. /3-lipotropin and /3-melanotropin did notaffect corticosterone production in decapsular cells.Although ACTH1-24 caused a significant increase incyclic AMPproduction in capsular cells in a dose-dependent fashion (1 nM-1 uM), 83-lipotropin and,8-melanotropin did not induce an increase in cyclicAMPproduction at the doses used (1 nM-I ,uM). The/3-melanotropin analogue (glycine[Gly]10-,8-melano-tropin) inhibited aldosterone production induced by ,B-lipotropin or /8-melanotropin, but did not inhibit aldo-sterone production induced by ACTH1-24 or angio-tensin II. Corticotropin-inhibiting peptide (ACTH7-38)inhibited not only ACTH1-24 action but also /lipotropinor 83-melanotropin action; however it did not affectangiotensin II-induced aldosterone production. (sara-lasin [Sar]i; alanine [Ala]8)-Angiotensin II inhibitedthe actions of 8-lipotropin and (3-melanotropin as wellas angiotensin II. These results indicate that (a) 38-lipotropin and ,8-melanotropin cause a significant stim-ulation of aldosterone production in capsular cells,(b) 3-lipotropin and 83-melanotropin have a preferentialeffect on zona glomerulosa cells, (c) (8-melanotropin
Dr. Matsuoka's present address is the University of Tokyo,Hongo, Bunkyo-ku, Tokyo 113, Japan.
Received for publication 4 December 1980 and in revisedform 20 April 1981.
752
contains the active peptide core necessary for aldo-sterone stimulation, (d) the effects of these peptideson aldosterone production may be independent ofcyclic AMP, and (e) the receptors for 3lipotropin or,3-melanotropin may be different from those for ACTHor angiotensin II.
INTRODUCTION
Although it is well recognized that ACTHand angio-tensin II have a potent aldosterone-stimulating effect,experimental evidence suggests that a non-ACTHpituitary factor(s) is important in the aldosteroneresponse during sodium restriction (1, 2). Also, clinicalevidence indicates the presence of an unknown aldo-sterone stimulator (3,4). Very recently we reported that,8-lipotropin induced aldosterone stimulation in ratadrenal capsular cells (5). In this study the mechanismof aldosterone-stimulating action of 8-lipotropin wasinvestigated. Also, because /8-lipotropin has the com-plete amino acid sequence of S-melanotropin and,3-endorphin (6-8), we investigated the effects ofthese /3-lipotropin-derived peptides on steroidogenesisusing rat adrenal collagenase-treated capsular anddecapsular cells.
tropin (l3-MSH), f-MSH analogue ([Gly1i]-f-MSH), andcorticotropin-inhibiting peptide (ACTH7-38; CIP) were syn-thesized according to reported methods (9-12). ACTH1-24was provided from Ciba-Geigy Corp. (Summit, N. J.), syn-thetic human 8-endorphin and synthetic methionine (Met)-and leucine (Leu)-enkephalin were purchased from Boeh-ringer Mannheim Biochemicals (Indianapolis, Ind.), syn-thetic (aspartic acid [Asp]'; isoleucine [Ile]5) angiotensin II
'Abbreviations used in this paper: AII, angiotensin II;,8-LPH, /8-lipotropin; /3-MSH, 3-melanotropin; CIP, cortico-tropin-inhibiting peptide.
(AII) and bovine serum albumin were obtained from SigmaChemical Co. (St. Louis, Mo.), and synthetic (Sarl, Ala8)angiotensin II (Saralasin) was from Calbiochem-BehringCorp. (San Diego, Calif.). Collagenase was from Worthing-ton Biochemical Corp. (Freehold, N. J.). 1-Methyl-3-isobutyl-xanthine was from Aldrich Chemical Co. (Milwaukee, Wis.).Medium 199 was from Gibco Laboratories (Grand IslandBiological Co., Grand Island, N. Y.).
Preparations of adrenal capsular and decapsular cells.20 Sprague-Dawley female rats (180-220 g) were used in eachexperiment. The rats were maintained on a regular Purina ratchow diet (Ralston Purina Co., St. Louis, Mo.) for at least2 wk. The rats were killed by decapitation and the adrenalglands were removed and separated into the capsular andthe decapsular portions according to reported methods (13).Each portion was incubated with 2 mg/ml collagenase anddispersed with a pasteur pipette. Collagenase-dispersedcapsular (mainly zona glomerulosa cells) and decapsular(zona fasciculata and reticularis cells) adrenal cells were usedfor the experiments.
Stimulation of steroidogenesis. The capsular and de-capsular cell suspensions (average cell counts: 100,000/tube)were incubated in duplicate in 1 ml of Medium 199 con-taining 2 mg/ml bovine serum albumin with various amounts(10 pM-3 ,uM) of synthetic f8-LPH, ,3-MSH, 8-endorphin,Met- and Leu-enkephalin, ACTH1-24, and AII for 2 h at 37°Cunder 95% 02 and 5%CO2.
Cyclic AMP stimulation. To define the mechanism ofaldosterone production by (3-LPH-derived peptides, thecapsular cell suspensions were incubted in duplicate in1 ml of Medium 199 containing 0.2 mM1-methyl-3-isobutyl-xanthine with various amounts (1 nM-1 ,M) of 18-LPH,,B-MSH, ACTH1-24, and AII for 1 h at 37°C. After incubation,700 Al of cell suspensions was transferred into glass tubescontaining 300 j1. of 2 mMtheophylline and boiled in awater bath for 10 min as described (14).
Competitive inhibition studies. To answer the question ofwhether 13-LPH and 13-MSH have their own distinct re-ceptors or not, the inhibition of aldosterone response tof8-LPH, j8-MSH, ACTH,1-24 and AII by the S3-MSH analogue,CIP, and Saralasin was examined. The capsular cell sus-pensions were preincubated with various amounts (0.1 nM-30 ,uM) of /8-MSH analogue, CIP, or Saralasin for 5 min andwere then incubated with submaximal amounts of f3-LPHand ,8-MSH (30 nM) and ACTH1-24 and AII (1.5 x 0.1 nM)for 2 h at 37°C.
Measurement of aldosterone, corticosterone, and cyclicAMP. Aldosterone in the incubation medium was measuredby a direct radioimmunoassay as described (13, 15). Thealdosterone value measured directly was well correlatedwith the value obtained after LH-20 column separation.(y = 1.1 x +3.5, r = 0.99, P < 0.001, n = 18). Corticosteronewas also measured directly by radioimmunoassay using ahighly specific antibody from Endocrine Science (Tarzana,Calif.). Cyclic AMPwas measured by protein-binding assayusing a kit from Amersham (Arlington Heights, Ill.) as de-scribed (16, 17). The interassay coefficients of variations forthe aldosterone, corticosterone, and cyclic AMPassays were10.0% (n = 12), 9.0% (n = 16), and 11.3% (n = 10), re-spectively. The intra-assay coefficients of variations for thealdosterone, corticosterone, and cyclic AMPassays were 6.3%(n = 16), 7.5% (n = 16), and 6.6% (n = 9), respectively. Therecoveries of aldosterone and corticosterone added to theincubation medium were 98.5+2.1% (SE) (n = 14) and91.6±1.5% (n = 16), respectively. The recovery of cyclicAMP added to the cell suspension before boiling was98.1±1.5% (n = 9). Statistical analysis was performed usingthe paired t test. A P value of <0.05 was considered significant.
RESULTS
Effects of f3-LPH-derived peptides, ACTH1-24, andAll on steroidogenesis. Significant increases in aldo-sterone production were produced by 0.1 nMACTH1-24or AII, and maximum increases were obtained with10 nM ACTH1-24 or AII in capsular cells. /3-LPH(10 nM) caused a significant increase in aldosteroneproduction (from the control value of 33.7±1.8 (SE)to 47.6±6.2 ng/100,000 capsular cells, P < 0.05). Maxi-mal aldosterone response was obtained with 3 x 0.1uM ,8-LPH (117.3±9.3 ng/100,000 capsular cells), and
the aldosterone level obtained with maximum doses of,8-LPH was similar to the one obtained with maximumdoses of All (Fig. 1). f3-LPH and 13-MSH caused sig-nificant stimulation of aldosterone, and their effects arecomparable (Fig. 2). 3-endorphin and Met- and Leu-enkephalin (data not shown) did not increase aldo-sterone production. Whereas ACTH1-24 has similareffects on capsular and decapsular cells, f8-LPH and(3-MSH did not affect corticosterone production at thedoses used in decapsular cells (Fig. 3). Neither AIInor ,-endorphin stimulated corticosterone productionin decapsular cells (Fig. 3).
Effects of f3LPH-derived peptides, ACTH1-24, andAll on cyclic AMPproduction. Although ACTH1-24
150
0 ~~~~~~~i24/U i ACTH%/
I /t AU\-P20,/ A
0 0.1nM 1n B-LP
I I
FIGURE 1 Effects of ACTH124, All, and (3-LPH on aldo-sterone production in capsular cells. Each point representsmean+SE from four different experiments. NS, not sig-nificantly different from the control value; *, significantlydifferent from the control value (P < 0.05). All other pointssignificantly different from the control value (P < 0.01).
Effects of f-Lipotropin Peptides on Aldosterone Production 753
C
o 100
U£UbO
a0a
0100
50C;
-
C
"I
0
00C0
4.
B-MSH
B-Endorphin
I nM 10nM 0.1jlM 1M
Peptide Concentration
10M
FIGURE 2 Effects of f-LPH-derived peptides on aldosteroneproduction in capsular cells. Each point represents mean±SE from three or four different experiments.
caused a significant cyclic AMPproduction in a dose-dependent fashion, ,3-LPH-derived peptides and Allat the doses used did not stimulate cyclic AMPpro-duction (Fig. 4). This failure to stimulate cyclic AMPwas present even at doses of /8-LPH and f8-MSH
S
SU
3*00006
000
50
0 200ko 15000 n";q
10 pM O. nM I nM 10nM 0.1gMPeptide Concentration
IjuM
FIGURE 3 Effects of ACTH'-24, AII, and f3-LPH-derivedpeptides on corticosterone production in decapsular cells.Each point represents mean+SE from three different experi-ments. *, significantly different from the control value(P < 0.01). All other points not significantly different fromthe control value.
-
* *30- **
o----o B-LPHS~~~~~~~~~~~~~~~~~~~~
CL o-aB-MSH
O 20- ** ACTHt 24
o T
a. 0
0.1 nb I ntA 10 nNI Wp1m 1 mm
Poptido Concntratlon
FIGURE4 Effects of ACTH1-24, ,B-LPH, ,8-MSH, and All oncyclic AMPproduction in capsular cells. Each point repre-sents mean+SE from three or four different experiments,*, **; significantly different from the control value (P < 0.05and P < 0.01, respectively). All other points not significantlydifferent from the control value.
(1 uM) that were larger than the doses required tostimulate aldosterone maximally (Figs. 2 and 4). Thedose of ACTH that stimulated aldosterone maximally(10 nM) doubled cyclic AMPproduction and the nexthigher dose (0.1 ,uM) tripled cyclic AMPproduction.Therefore, the pattern of cyclic AMP response to,3-LPH and (8-MSH is different from the pattern afterACTHand similar to the pattern after AII.
Inhibition of aldosterone production by f3MSHanalogue, CIP, or Saralasin. The effects of the ,8-MSHanalogue on the aldosterone responses to submaxi-mal-stimulating doses of /8-LPH or f8-MSH (30 nM)and of ACTH1-24 or All (0.15 nM) are shown in Fig.5. These doses of peptides gave comparable in-creases in aldosterone production, yet the productionrates were still submaximal (Tables I, II). The f3-MSHanalogue was a potent inhibitor of aldosterone pro-duction induced by f3-LPH and 8-MSH, causing -30%inhibition at a molar ratio of 100:1 and -80% inhibitionat a molar ratio of 1,000: 1. The f8-MSH analogue, how-ever, did not affect ACTH1-24 or AII-induced aldo-sterone production at molar ratios from 10:1 to 10,000:1.On the other hand, CIP inhibited aldosterone pro-duction induced not only by ACTH1-24 but also by,8-LPH or ,f-MSH (Fig. 6, Table II). In one experimenta large dose of 1-LPH (1 ,uM) was used. CIP, in molarratios of 1:1 and 10:1, inhibited the aldosterone re-sponse by 20 and 25%, respectively. The control aldo-sterone production was 26.5 ng/105 adrenal cells, androse to 146 ng/105 cells with 1 uM 3-LPH. CIP at
754 H. Matsuoka, P. J. Mulrow, R. Franco-Saenz, and C. H. Li
c0
0
000.CL0S
0
00.
100-
50 -
A- - -
ACTH'24
S
'\ B-MSH
B-LPH &.
110 100 1,000 10,000
Molar Ratio; B-MSH Analog/peptide
FIGURE 5 Inhibition of aldosterone responses to submaximalstimulating doses of f3-LPH and 8-MSH (30 nM), andACTH'-24 and All (0.15 nM) by various concentrations of,8-MSH analogue. Each point represents mean+SE fromthree different experiments.
1 ,tM reduced the response to 122 ng and at 10 ,uM re-duced the response to 102 ng. Although ACTHat 10 nMcaused a similar aldosterone response (145 ng), CIPat molar ratios of 1:1 and 10:1 had no inhibitory effect.It would appear, therefore, that 8-LPH is more sensi-tive to the inhibitory action of CIP than is ACTH.CIP, however, did not inhibit aldosterone productioninduced by All. Also, Saralasin inhibited not only theAll-induced aldosterone production but also the ,3-LPH- and ,B-MSH-induced aldosterone production.However, this analogue did not affect aldosterone pro-duction induced by ACTH1-24 (Fig. 7). The J3-MSHanalogue, CIP, and Saralasin had no significant aldo-sterone-stimulating activity in capsular cells at dosesfrom 1 nM to 30 ,tM (unpublished data).
DISCUSSION
There is considerable evidence indicating that theregulation of aldosterone secretion cannot be com-pletely explained by the known stimulators. Palmoreand Mulrow (1) and McCaa et al. (2) have reported thata non-ACTH pituitary factor(s) is essential for the aldo-sterone response during sodium restriction in rats anddogs. Recently, Sen et al. (18) reported the isolationof a glycoprotein from normal human urine that pro-duced hypertension associated with high plasma aldo-sterone values in the rat. Removal of some carbo-hydrate moieties with neurominadase increased the
activity of this protein on aldosterone production invitro (19). In view of these reports, we have ex-amined the effects of 8-LPH-derived peptides onsteroidogenesis using rat adrenal glands. Although,3-LPH is derived from a common pro-ACTH/,B-LPH-glycosylated precursor molecule present in the pitui-tary gland (6, 7), its physiological role remains un-clear. ,-LPH (1-91 amino acids) contains within it theentire amino acid sequence of 8-MSH (,8-LPH41-58)(6, 8). In our studies ,B-LPH and 3-MSH have a sig-nificant aldosterone-stimulating effect and similar toAll have a preferential effect on the zona glomerulosacells. Because f3-endorphin (,8-LPH61-91) has nosteroidogenic property, the significant portion of theactive peptide core necessary for aldosterone stimula-tion is included in j-MSH. 8-MSH contains the hepta-peptide sequence (Met-glutamic acid(Glu)-histidine-(His)-phenylalanine(Phe)-arginine(Arg)-tryptophan-(Trp)-glycine(Gly)) common to ACTH, and 8-LPH whichmay be the peptide core required for aldosteronestimulation. Previous studies suggest that ,8-MSH hasonly weak steroidogenic potency on the adrenal glandin the rat (20, 21). However, in those studies aldo-sterone production was not examined. It has been re-
TABLE IInhibition of Aldosterone Production by -MSH Analogue
ported that human (8-MSH may be a degradationproduct of f3-LPH during extraction of plasma orpituitary gland and that in human plasma the immuno-assayable f3-MSH may be fB-LPH (22, 23). In somespecies, however, the pars intermedia may be a mainsource of 8-MSH (24). The secretion of ACTH and,8-LPH or f8-MSH occurs simultaneously in most in-stances. Several investigators, however, reporteddissociation of ACTHand 13-MSH (or ,B-LPH) secre-tion under some conditions (25-29). Interestingly,Howeand Thody (30) reported that changes in sodiumbalance induced changes in the content of MSHas wellas histological changes in the pars intermedia of therat. Also, Kobayashi and Takema (31) reported histo-logical changes in the pars intermedia of mice duringsodium restriction. Furthermore, Page et al. (32)demonstrated that the concomitant administration ofa-MSH and growth hormone stimulated aldosteronesecretion in hypophysectomized sodium-depleted rats,while Vinson et al. (33) reported that a-MSH stimulatedaldosterone production by adrenal glomerulosa cellsin vitro. A synthetic peptide, y3-MSH, which representsa portion of the amino terminal or 16K fragment withinthe pro-ACTH/,8-LPH precursor molecule, potentiatesthe stimulation of aldosterone production by ACTH
100-C0U0a
h.0c0
* 00
C0
0A.
B-MSH I
AZ
, ACTH'-24
100 1,000 10,000 100,000
Molar Ratio; CIP/peptide
FIGURE 6 Inhibition of aldosterone responses to submaximalstimulating doses of 13-LPH and 13-MSH (30 nM), andACTH1-24 and AII (0.15 nM) by various concentrations ofCIP. Each point represents meantSE from three differentexperiments.
in hypophysectomized rats (34). These authors specu-late that the y-MSH region of the 16K fragment mayplay a role in control of steroidogenesis. These datasuggest that /8-MSH or other similar pituitary pep-
.; 1.>>,,ACTH 1 l2
0100 -
OB-LPHM\ '-
C~~~~~~~~~\ B-MSH
o \
~50-
U~~~A
9Li
10 100 1,000 10,000 100,000
Molar Ratio; Saralasin/peptide
FIGURE 7 Inhibition of aldosterone responses to sub-maximal stimulating doses of P-LPH and ,8-MSH (30 nM), andACTH'-24 and All (0.15 nM) by various concentrations ofSaralasin. Each point represents mean±SE from three differ-ent experiments.
756 H. Matsuoka, P. J. Mulrow, R. Franco-Saenz, and C. H. Li
tides may be involved in the regulation of aldosteronesecretion.
It has been shown that 13-LPH increases the level ofplasma aldosterone in the rat in vivo (35). In this study,3-LPH and f3-MSH have a preferential effect on thezona glomerulosa cells. These results raise the pos-sibility that f3-LPH or other pituitary peptides play arole in the regulation of aldosterone secretion. Therelatively high doses needed to see an effect on aldo-sterone production militate against this possibility.In our studies, the smallest concentration of j-LPHthat frequently stimulated aldosterone slightly (al-though not statistically significant) was 1 nM/liter.This concentration is - 100 times higher than the levelsfound in normal human peripheral plasma (36, 37).However, in patients with Nelson's syndrome (-2x 1 nM) and Addison's disease (-2 x 0.1 nM), ,3-LPHlevels approach this concentration (36,37). The plasmalevels of,f-LPH in conditions with hyperaldosteronismare unknown. Furthermore, in our study we investi-gated the effects of ovine f3-LPH and camel ,8-MSH inthe rat and compared their effects against ACTH1-24,which is known to be the biologically active sequenceof ACTH for all species. Species differences havebeen shown for the lipolytic effect of B-LPH (38, 39).Therefore it may be possible that species differencemay exist in the aldosterone response to ,3-LPH-derived peptides. It may also be possible that 8-LPHor f3-MSH is a component of a larger pituitary peptidewhich may be more potent in vitro or in vivo. Finally,the adrenal gland may well develop increased sensi-tivity to (8-LPH under certain conditions, as it does toAII following sodium depletion and to ACTH inCushing's disease.
To elucidate the mechanism of the aldosterone-stimulating activity of f3-LPH and f8-MSH, the cyclicAMP responses to ACTH1-24 and f8-LPH-derivedpeptides were examined. The concept that cyclicAMP is the mediator of the steroidogenic action ofACTH on the adrenal gland has been supported byprevious studies (40,41). Fujita et al. (14) have reportedthat the action of ACTH on aldosterone secretion ismediated by cyclic AMP. In their study as well as inours, the concentration of ACTH (1 nM) required tostimulate a significant cyclic AMPproduction is 10times higher than that required to induce a significantaldosterone stimulation. (3-LPH and f8-MSH at thedoses used stimulated aldosterone production withoutcausing detectable changes in cyclic AMPproduction.In this respect, /3-LPH and f8-MSH may act on aldo-sterone secretion through a mechanism similar tothat of All rather than of ACTH, because even maxi-mumaldosterone-stimulating doses of f8-LPH (1 ,M)did not increase cyclic AMP. Several investigatorshave suggested that only a small increase of cyclicAMPproduction is necessary to induce steroidogenesis
(42, 43). If this is the case, we cannot exclude the roleof cyclic AMPin the aldosterone stimulation inducedby ,8-LPH and 1B-MSH.
The 8-MSH analogue inhibited aldosterone pro-duction induced by 83-LPH or ,3-MSH to a similardegree. However, aldosterone production induced byACTH1-24 or AII was not affected by the ,B-MSHanalogue. These results suggest that the receptors for,8-LPH are the same as those for,8-MSH and may bedifferent from those for ACTHor AII. It is known thatCIP has a 32-amino acid sequence that correspondsto the ACTH7-38 sequence and is isolated from humanpituitary glands (12). This peptide is devoid of corti-costeroidogenic activity and causes a 50% inhibitionof ACTH-induced corticosterone production by ratadrenal cells in vitro at a molar ratio of --10,000:1(12). Our finding with respect to aldosterone inhibitionis quite similar. In this study CIP inhibited not onlyACTHbut also ,B-LPH and ,-MSH action and appearsto be a more effective inhibitor of the latter peptides.CIP, however, did not inhibit All action. These resultssuggest that there may be two kinds of binding sitesfor ACTH with different affinities, and one of thesebinding sites may be shared by B-LPH-derived pep-tides. Lefkowitz et al. (44) also proposed the existenceof two kinds of binding sites for ACTH in a mouseadrenal cell tumor. As an alternative explanation, CIPmay interact with the peptides and cause structuralchanges that modify binding to the receptor. It isproposed that a specific conformation of ACTH isnecessary for receptor binding (45).
The effect of Saralasin on aldosterone productioninduced by ,B-LPH or ,B-MSH is difficult to explain.Saralasin inhibited AII, f-LPH, and f8-MSH actionsbut did not affect ACTHaction. One possible explana-tion would be that the receptor for 8-LPH-derivedpeptides shares some binding properties with the AIIand ACTH receptors. However, it is unlikely that3-LPH-derived peptides and AII share the same re-
ceptors, because their amino acid sequence are com-pletely different. Another possibility is that high dosesof Saralasin cause changes at the receptor level similarto those reported after the addition of guanylnucleo-tides (46). In our study CIP did not inhibit AII action,and Saralasin did not inhibit ACTHaction. Therefore,it is unlikely that the effect of high doses of CIP andSaralasin is a toxic or nonspecific effect. Furtherstudies are necessary to resolve these problems.
In conclusion 8-LPH and 8-MSH stimulate aldo-sterone production in capsular cells. In contrast toACTH, 8-LPH- and f3-LPH-derived peptides do notaffect cyclic AMPproduction in capsular cells and donot increase corticosterone production in decapsularcells, suggesting that the mechanism of action may bedifferent from that of ACTH. These peptides may play arole in the regulation of aldosterone secretion.
Effects of /3-Lipotropin Peptides on Aldosterone Production 757
ACKNOWLEDGMENT
This work was supported in part by National Institutes ofHealth grant H1-19644 and grant GM-2907.
REFERENCES
1. Palmore, W. P., and P. J. Mulrow. 1967. Control of aldo-sterone secretion by the pituitary gland. Science (Wash.,D. C.). 158: 1482-1484.
2. McCaa, R. E., D. B. Young, A. C. Guyton, and C. S. McCaa.1974. Evidence for a role of an unidentified pituitaryfactor in regulating aldosterone secretion during alteredsodium balance. Circ. Res. Suppl. 34-35: 15-25.
3. Davis, W. W., H. H. Newsome, L. D. Wright, Jr., W. G.Hammond, J. Easton, and F. C. Bartter. 1967. Bilateraladrenal hyperplasia as a cause of primary aldosteronismwith hypertension, hypokalemia and suppressed reninactivity. Am. J. Med. 42: 642-647.
4. Nicholls, M. G., E. A. Espiner, H. Hughes, J. Ross, andD. T. Stewart. 1975. Primary aldosteronism; a study incontrasts. Am. J. Med. 59: 334-342.
5. Matsuoka, H., P. J. Mulrow, and C. H. Li. 1980. /3-lipo-tropin; a new aldosterone-stimulating factor. Science(Wash. D. C.). 209: 307-308.
6. Nakanishi, S., A. Inoue, T. Kita, M. Nakamura, A. C. Y.Chang, S. N. Cohen, and S. Numa. 1979. Nucleotidesequence of cloned cDNA for bovine corticotropin-,3-lipotropin precursor. Nature (Lond.). 278: 423-427.
7. Imura, H., and Y. Nakai. 1981. "Endorphins" in pituitaryand other tissues. Annu. Rev. Physiol. 43: 265-278.
8. Li, C. H., L. Bamafi, M. Chretien, and D. Chung. 1965.Isolation and amino-acid sequence of ,8-LPH from sheeppituitary glands. Nature (Lond.). 208: 1093-1094.
9. Yamashiro, D., and C. H. Li. 1978. Total synthesis ofovine f3-lipotropin by the solid-phase method. J. Am.Chem. Soc. 100: 5174-5179.
10. Li, C. H., D. Yamashiro, and S. Lemaire. 1975. Totalsynthesis of camel 8-melanotropin by the solid-phasemethod. Biochemistry. 14: 953-956.
11. Lemaire, S., D. Yamashiro, A. J. Rao, and C. H. Li. 1977.Synthesis and biological activity of /8-melanotropins andanalogues. J. Med. Chem. 20: 155-158.
12. Li, C. H., D. Chung, D. Yamashiro, and C. Y. Lee. 1978.Isolation, characterization, and synthesis of a cortico-tropin inhibiting peptide from human pituitary glands.Proc. Natl. Acad. Sci. U. S. A. 75: 4306-4309.
13. Matsuoka, H., S. Y. Tan, and P. J. Mulrow. 1980. Effects ofprostaglandins on adrenal steroidogenesis in the rat.Prostaglandins. 19: 291-298.
14. Fujita, K., G. Aguilera, and K. J. Catt. 1979. The role ofcyclic AMPin aldosterone production by isolated zonaglomerulosa cells. J. Biol. Chem. 254: 8567-8574.
15. Tan, S. Y., R. Noth, and P. J. Mulrow. 1978. Direct non-chromatographic radioimmunoassay of aldosterone:validation of a commercially available kit and observa-tions on age-related changes in concentrations in plasma.Clin. Chem. 24: 1531-1533.
16. Gilman, A. G. 1970. A protein binding assay for adenosine3':5'-cyclic monophosphate. Proc. Natl. Acad. Sci.U. S. A. 67: 305-312.
17. Tovey, K. C., K. G. Oldham, and J. A. M. Whelan. 1974.A simple direct assay for cyclic AMP in plasma andother biological samples using an improved competitiveprotein binding technique. Clin. Chim. Acta. 56: 221-234.
18. Sen, S., E. L. Bravo, and F. M. Bumpus. 1977. Isolationof a hypertension-producing compound from normalhuman urine. Circ. Res. Suppl. 40: 5-10.
19. Sen, S., J. R. Shainoff, E. L. Bravo, and F. M. Bumpus.1981. Isolation of aldosterone-stimulating factor (ASF)and its effect on rat adrenal glomerulosa cells in vitro.Hypertension (Dallas). 3: 4-10.
20. Sayers, G., R. L. Swallow, and N. D. Giordano. 1971.An improved technique for the preparation of isolatedrat adrenal cells: a sensitive, accurate and specific methodfor the assay of ACTH. Endocrinology 88: 1063-1068.
21. Lowry, P. J., C. McMartin, and J. Peters. 1973. Propertiesof a simplified bioassay for adrenocorticotropic activityusing the steroidogenic response of isolated adrenalcells. J. Endocrinol. 59: 43-55.
22. Bloomfield, G. A., A. P. Scott, P. J. Lowry, J. J. H. Gilkes,and L. H. Rees, 1974. A reappraisal of human 8-MSH.Nature (Lond.). 252: 492-493.
23. Gilkes, J. J. H., G. A. Bloomfield, A. P. Scott, P. J. Lowry,J. G. Ratcliffe, J. Landon, and L. H. Rees. 1975. Develop-ment and validation of a radioimmunoassay for peptidesrelated to /3-melanocyte-stimulating hormone in humanplasma: the lipotropins. J. Clin. Endocrinol. Metab.40: 450-457.
24. Lowry, P. J., and A. P. Scott. 1975. The evolution of verte-brate corticotrophin and melanocyte stimulating hor-mone. Gen. Comp. Endocrinol. 26: 16-23.
25. Abe, K., W. E. Nicholson, G. W. Liddle, D. N. Orth, andD. P. Island. 1969. Normal and abnormal regulation of,B-MSH in man.J. Clin. Invest. 48: 1580-1585.
26. Kastin, A. J., A. V. Schally, S. Viosca, and M. C. Miller.1969. MSHactivity in plasma and pituitaries of rats aftervarious treatments. Endocrinology. 84: 20-27.
27. Dunn, J. D., A. J. Kastin, A. J. Carrillo, and A. V. Schally.1972. Additional evidence for dissociation of melanocyte-stimulating hormone and corticotrophin release. J. Endo-crinol. 55: 463-464.
28. Kastin, A. J., G. D. Beach, W. D. Hawley, J. W. Kendall,Jr., M. S. Edwards, and A. V. Schally. 1973. Dissociationof MSHand ACTHrelease in man. J. Clin. Endocrinol.Metab. 36: 770-772.
29. Hirata, Y., N. Sakamoto, S. Matsukura, and H. Imura.1975. Plasma levels of 18-MSH and ACTHduring acutestresses and metyrapone administration in man. J. Clin.Endocrinol. Metab. 41: 1092-1097.
30. Howe, A., and A. J. Thody. 1970. The effect of ingestion ofhypertonic saline on the melanocyte-stimulating hormonecontent and histology of the pars intermedia of the ratpituitary gland. J. Endocrinol. 46: 201-208.
31. Kobayashi, Y., and M. Takema. 1976. A morphometricstudy on the pars intermedia of the hypophysis duringimpairment of the renin-angiotensin-aldosterone systemin sodium depleted mice. Cell Tissue Res. 168: 153-159.
32. Page, R. B., J. E. Boyd, and P. J. Mulrow. 1974. Theeffect of alpha-melanocyte stimulating hormone on aldo-sterone production in the rat. Endocr. Res. Commun.1: 53-62.
33. Vinson, G. P., B. J. Whitehouse, A. Dell, T. Etienne,and H. R. Morris. 1980. Characterization of an adrenalzona glomerulosa-stimulating component of posteriorpituitary extracts as Alpha-MSH. Nature (Lond.). 284:464-467.
34. Pedersen, R. C., A. C. Brownie, and N. Ling. 1980.Pro-adrenocorticotropin/endorphin-derived peptides:coordinate action on adrenal steroidogenesis. Science(Wash. D. C.). 208: 1044-1045.
35. Matsuoka, H., P. J. Mulrow, R. Franco-Saenz, and C. H. Li.1980. Effects of 3-lipotropin on aldosterone productionin rats. Clin. Sci. (Lond.). 59(Suppl. 6): 91S-94S.
36. Wiedemann, E., T. Saito, J. A. Linfoot, and C. H. Li. 1977.
758 H. Matsuoka, P. J. Mulrow, R. Franco-Saenz, and C. H. Li
Radioimmunoassay of human ,-lipotropin in unextractedplasma. J. Clin. Endocrinol. Metab. 45: 1108-1111.
37. Krieger, D. T., A. S. Liotta, T. Suda, A. Goodgold, and E.Condon. 1979. Humanplasma immunoreactive lipotropinand adrenocorticotropin in normal subjects and in patientswith pituitary-adrenal disease. J. Clin. Endocrinol.Metab. 48: 566-571.
38. Lohmar, P., and C. H. Li. 1968. Biological properties ofovine ,-lipotropic hormone. Endocrinology. 82: 898-904.
39. Lis, M., C. Gilardeau, and M. Chretien. 1972. Fat celladenylate cyclase activation by sheep 8-lipotropic hor-mone. Proc. Soc. Exp. Biol. Med. 139: 680-683.
40. Haynes, R. C., S. B. Koritz, and F. G. Peron. 1959. In-fluence of adenosine 3',5'-monophosphate on corticoidproduction by rat adrenal glands. J. Biol. Chem. 234:1421- 1423.
41. Grahame-Smith, D. G., R. W. Butcher, R. L. Ney, andE. W. Sutherland. 1967. Adenosine 3',5'-monophosphateas the intracellular mediator of the action of adreno-
corticotropic hormone on the adrenal cortex. J. Biol.Chem. 242: 5535-5541.
42. Beall, R. J., and G. Sayers. 1972. Isolated adrenal cells:steroidogenesis and cyclic AMPaccumulation in responseto ACTH. Arch. Biochem. Biophys. 148: 70-76.
43. Schimmer, B. P., and A. E. Zimmerman. 1976. Steroido-genesis and extracellular cAMPaccumulation in adrenaltumor cell cultures. Mol. Cell. Endocr. 4: 263-270.
44. Lefkowitz, R. J., J. Roth, and I. Pastan. 1971. ACTH-receptor interaction in the adrenal: a model for the initialstep in the action of hormones that stimulate adenylcyclase. Ann. N. Y. Acad. Sci. 185: 195-209.
45. Seelig, S., G. Sayers, R. Schwyzer, and P. Schiller. 1971.Isolated adrenal cells: ACTH"I-24, a competitive an-tagonist of ACTH-39 and ACTH'-"0. Febs (Fed. Eur.Biochem. Soc.) Lett. 19: 232-234.
46. Glossmann, H., A. Baukal, and K. J. Catt. 1974. Angio-tensin II receptors in bovine adrenal cortex: modificationof angiotensin II binding by guanylnucleotides. J. Biol.Chem. 249: 664-666.
Effects of f3Lipotropin Peptides on Aldosterone Production 759
![Clinical data successes - Joseph Paul Cohen...cat = [0 0 1 0 0 0 0 0 0 0 0 0 0 0 … 0] dog = [0 0 0 0 1 0 0 0 0 0 0 0 0 0 … 0] house = [1 0 0 0 0 0 0 0 0 0 0 0 0 0 … 0] Note!](https://static.documents.pub/doc/80x56/5fdf222a2dd17b0d95129a68/clinical-data-successes-joseph-paul-cohen-cat-0-0-1-0-0-0-0-0-0-0-0-0-0.jpg)
![[XLS] 7-10... · Web view1 0 0 0 2 0 0 0 3 0 0 0 4 0 0 0 5 0 0 0 6 0 0 0 7 0 0 0 8 0 0 0 9 0 0 0 10 0 0 0 11 0 0 0 12 0 0 0 13 0 0 0 14 0 0 0 15 0 0 0 16 0 0 0 17 0 0 0 18 0 0 0 19](https://static.documents.pub/doc/80x56/5ae8a6607f8b9a29049069b5/xls-7-10web-view1-0-0-0-2-0-0-0-3-0-0-0-4-0-0-0-5-0-0-0-6-0-0-0-7-0-0-0-8-0.jpg)
