The intermediate lobe of rodent pituitaries is involved in the regulation of prolactin (PRL) secretion from the anterior lobe. In a previous study, we demonstrated the stimulatory effect of α-melanocyte-stimulating hormone (α-MSH) on PRL release and the expression of melanocortin-3 receptors (MC3-Rs) in cultured mouse pituitary cells. The aim of the present study was to clarify whether α-MSH directly stimulates PRL release through the MC3-Rs by determining the cell type of MC3-R-expressing cells in the mouse pituitary anterior lobe. Northern blot analysis revealed a 2.7-kb transcript for MC3-R mRNA in the anterior and neurointermediate lobes of pituitary glands of adult male and female mice. Dual cellular localization of MC3-R mRNA and PRL or growth hormone (GH) in the mouse pituitary glands was performed by in situ hybridization analysis of MC3-R mRNA followed by immunocytochemical detection of PRL or GH. MC3-R mRNA was detected in most mammotropes and some somatotropes. α-MSH increased PRL release and stimulated DNA replication in mammotropes, and these effects were blocked by SHU9119, an antagonist of MC3-R and MC4-R. These results indicate that α-MSH stimulates PRL release and proliferation of mammotropes through MC3-Rs, and suggest that α-MSH from intermediate lobes can regulate mammotrope functions in the mouse pituitary.

1.
Ben-Jonathan N: Regulation of prolactin secretion; in Imura H (ed): The Pituitary Gland. New York, Raven Press, 1994, pp 261–283.
2.
Hill JB, Nagy GM, Frawley LS: Suckling unmasks the stimulatory effect of dopamine on prolactin release: Possible role for α-melanocyte-stimulating hormone as a mammotrope responsiveness factor. Endocrinology 1991;129:843–847.
3.
Hill JB, Lacy ER, Nagy GM, Gorcs TJ, Frawley LS: Does α-melanocyte-stimulating hormone from the pars intermedia regulate suckling-induced prolactin release? Supportive evidence from morphological and functional studies. Endocrinology 1993;133:2991–2997.
4.
Murai I, Ben-Jonathan N: Acute stimulation of prolactin release by estradiol: Mediation by the posterior pituitary. Endocrinology 1990;126:3179–3184.
5.
Ellerkmann E, Nagy GM, Frawley LS: Rapid augmentation of prolactin cell number and secretory capacity by an estrogen-induced factor released from the neurointermediate lobe. Endocrinology 1991;129:838–842.
6.
Ellerkmann E, Nagy GM, Frawley LS: α-Melanocyte-stimulating hormone is a mammotrophic factor released by neurointermediate lobe cells after estrogen treatment. Endocrinology 1992;130:133–138.
7.
Morooka Y, Oomizu S, Takeuchi S, Takahashi S: Augmentation of prolactin release by α-melanocyte-stimulating hormone is possibly mediated by melanocortin 3-receptors in the mouse anterior pituitary cells. Zool Sci 1998;15:567–572.
8.
Cone RD, Lu D, Koppula S, Vage DI, Klungland H, Boston B, Chen W, Orth DN, Pouton C, Kesterson RA: The melanocortin receptors: Agonists, antagonists, and the hormonal control of pigmentation. Recent Prog Horm Res 1996;51:287–317.
9.
Roudbaraki M, Lorsignol A, Langouche L, Callewaert G, Vankelecom H, Denef C: Target cells of γ3-melanocyte-stimulating hormone detected through intracellular Ca2+ responses in immature rat pituitary constitute a fraction of all main pituitary cell types, but mostly express multiple hormone phenotypes at the messenger ribonucleic acid level. Refractoriness to melanocortin-3 receptor blockade in the lacto-somatotroph lineage. Endocrinology 1999;140:4874–4885.
10.
Takahashi S, Kawashima S: Proliferation of prolactin cells in the rat: Effects of estrogen and bromocryptine. Zool Sci 1987;4:855–860.
11.
Hruby VJ, Lu D, Sharma SD, Castrucci AL, Kesterson RA, al-Obeidi FA, Hadley ME, Cone RD: Cyclic lactam α-melanotropin analogues of Ac-N1e4-cyclo [Asp5, D-Phe7,Lys10] α-melanocyte-stimulating hormone-(4–10)-NH2 with bulky aromatic amino acids at position 7 show high antagonist potency and selectivity at specific melanocortin receptors. J Med Chem 1995;38:3454–3461.
12.
Schioth HB, Muceniece R, Mutulis F, Bouifrouri AA, Mutule I, Wikberg JE: Further pharmacological characterization of the selective melanocortin 4 receptor antagonist HS014:Comparison with SHU9119. Neuropeptides 1999;33:191–196.
13.
Chomczynski P, Sacchi N: Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 1987;162:156–159.
14.
Takeuchi S, Takahashi S: A possible involvement of melanocortin 3 receptor in the regulation of adrenal gland function in the chicken. Biochim Biophys Acta 1999;1448:512–518.
15.
Takahashi S: Growth hormone secretion in old female rats analyzed by the reverse hemolytic plaque assay. Acta Endocrinol 1992;127:531–535.
16.
Honda J, Oomizu S, Kiuchi Y, Komatsu N, Takeuchi S, Takahashi S: Identification of epidermal growth factor mRNA-expressing cells in the mouse anterior pituitary. Neuroendocrinology 2000;71:155–162.
17.
Oomizu S, Takahashi S: Insulin stimulates the proliferation of mouse anterior pituitary cells in vitro. Biomed Res 1996;17:365–371.
18.
Oomizu S, Takeuchi S, Takahashi S: Stimulatory effect of insulin-like growth factor I on proliferation of mouse pituitary cells in serum-free culture. J Endocrinol 1998;157:53–62.
19.
Hsu SM, Soban E: Color modification of diaminobenzidine precipitation by metallic ions and its application for double immunohistochemistry. J Histochem Cytochem 1982;30:1079–1082.
20.
Hsu SM, Raine L, Fanger H: Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: A comparison between ABC and unlabeled antibody (PAP) procedures. J Histochem Cytochem 1981;29:577–580.
21.
Roselli-Rehfuss L, Mountjoy KG, Robbins LS, Mortrud MT, Low MJ, Tatro JB, Entwistle ML, Simerly RB, Cone RD: Identification of a receptor for α-melanotropin and other proopiomelanocortin peptides in the hypothalamus and limbic system. Proc Natl Acad Sci USA 1993;90:8856–8860.
22.
Desarnaud F, Labbé O, Eggerickx D, Vassart G, Parmentier M: Molecular cloning, functional expression and pharmacological characterization of a mouse melanocortin receptor gene. Biochem J 1994;299:367–373.
23.
Takahashi S: Development and heterogeneity of prolactin cells. Int Rev Cytol 1995;157:33–98.
24.
Zheng T, Villalobos C, Nusser KD, Gettys TW, Faught WJ, Castaña JP, Frawley LS: Phenotypic characterization and functional correlation of α-MSH binding to pituitary cells. Am J Physiol 1997;272:E282–E287.
25.
Nuñez L, Frawley LS: α-MSH potentiates the responsiveness of mammotropes by increasing Ca2+ entry. Am J Physiol 1998;274:E971–E977.
26.
Ellerkmann E, Porter TE, Nagy GM, Frawley LS: N-acetylation is required for the lactotrope recruitment activity of α-melanocyte-stimulating hormone and β-endorphin. Endocrinology 1992;131:566–570.
27.
Laudon M, Grossman DA, Ben-Jonathan N: Prolactin-releasing factor: Cellular origin in the intermediate lobe of the pituitary. Endocrinology 1990;126:3185–3192.
28.
Allen DL, Low MJ, Allen RG, Ben-Jonathan N: Identification of two classes of prolactin-releasing factors in intermediate lobe tumors from transgenic mice. Endocrinology 1995;136:3093–3099.
29.
Lindley SE, Gunnet JW, Lookingland KJ, Moore KE: Effects of alterations in the activity of tuberohypophysial dopaminergic neurons on the secretion of α-melanocyte-stimulating hormone. Proc Soc Exp Biol Med 1988;188:282–286.
30.
Benjannet S, Rondeau N, Day R, Chrétien M, Seidah NG: PC1 and PC2 are proprotein convertases capable of cleaving proopiomelanocortin at distinct pairs of basic residues. Proc Natl Acad Sci USA 1991;88:3564–3568.
31.
Marcinkiewicz M, Day R, Seidah NG, Chretien M: Ontogeny of the prohormone convertases PC1 and PC2 in the mouse hypophysis and their colocalization with corticotropin and α-melanotropin. Proc Natl Acad Sci USA 1993;90:4922–4926.
32.
Nagy GM, Boockfor FR, Frawley LS: The suckling stimulus increases the responsiveness of mammotropes located exclusively within the central region of the adenohypophysis. Endocrinology 1991;128:761–764.
33.
Porter TE, Frawley LS: Neurointermediate lobe peptides recruit prolactin-secreting cells exclusively within the central region of the adenohypophysis. Endocrinology 1992;131:2649–2652.
34.
Murakami T, Ohtsuka A, Taguchi T, Kikuta A, Ohtani O: Blood vascular bed of the rat pituitary intermediate lobe, with special reference to its development and portal drainage into the anterior lobe. A scanning electron microscope study of vascular casts. Arch Histol Jpn 1985;48:69–87.
35.
Tilemans D, Ramaekers D, Andries M, Denef C: Effect of POMC1–76, its C-terminal fragment γ3-MSH and anti-POMC1–76 antibodies on DNA replication in lactotrophs in aggregate cell cultures of immature rat pituitary. J Neuroendocrinol 1997;9:627–637.
36.
Bert C, Vande Vijver V, Andries M, Verhaert P, Proost P, De Vreese B, Van Beeumen J, Vankelecom H, Denef C: Production of recombinant rat proopiomelanocortin 1–74 and characterization of its mitogenic action on pituitary lactotrophs. Mol Cell Endocrinol 1999;154:111–122.
37.
Langouche L, Roudbaraki M, Pals K, Denef C: Stimulation of intracellular free calcium in GH3 cells by γ3-melanocyte-stimulating hormone. Involvement of a novel melanocortin receptor? Endocrinology 2001;142:257–266.
38.
Langouche L, Pals K, Denef C: Structure-activity relationship and signal transduction of γ-MSH peptides in GH3 cells: Further evidence for a new melanocortin receptor. Peptides 2002;23:1077–1086.
39.
Oomizu S, Honda J, Takeuchi S, Kakeya T, Masui T, Takahashi S: Transforming growth factor-α stimulates proliferation of mammotrophs and corticotrophs in the mouse pituitary. J Endocrinol 2000;165:493–501.
40.
Denef C: Paracrine mechanisms in the pituitary; in Imura H (ed): The Pituitary Gland. New York, Raven Press, 1994, pp 351–378.
41.
Schwartz J: Intercellular communication in the anterior pituitary. Endocr Rev 2000;21:488–513.
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