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Intragranular processing of pro-opiomelanocortin in the intermediate cells of the rat pituitary glands

A quantitative immunocytochemical approach

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Summary

Quantitative immunocytochemical studies were done by using the immunogold technique on sections of the intermediate lobe of rat pituitary. Antibodies raised (in rabbits) against the precursor proteins pro-opiomelanocortin (POMC) and ACTH were used. The results clearly indicate that the immature granules are the major site of POMC, as their antigenic density (gold beads/μm2) was almost 3 times as high as that of ACTH. In the mature igranules, the antigenic density of ACTH was increased by 2.7-fold compared with the immature granules. Using a computer-assisted method, it was possible to categorize the granules antigenic density according to their size. Using this approach it was found that the antigenic density of POMC remained constant in all mature granules of varied sizes, whereas the antigenic density of ACTH decreased with increasing granule size. The relationship between granule size, degree of maturation, and antigenic density is discussed.

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References

  • Bendayan M, Zollinger M (1983) Ultrastructural localization of antigenic sites on osmium-fixed tissues applying the protein A gold technique. J Histochem Cytochem 31:101–109

    Google Scholar 

  • Cantin M, Ballak M, Lu CL, Seidah NG, Chrétien M (1983) Corticotropin (ACTH) and the N-terminal fragment of pro-opiomelanocortin are located in the same granules in cells of rat pituitary gland. J Histochem Cytochem 31:479–487

    Google Scholar 

  • Case MR (1978) Formation, intracellular transport and discharge of exportable proteins in pancreatic acinar cells and other cells. Biol Rev 53:211–354

    Google Scholar 

  • Chang T-L, Loh YP (1984) In vitro processing of proopiocortin by membrane-associated and soluble converting enzyme activities from rat intermediate lobe secretory granules. Endocrinology 114:2092–2099

    Google Scholar 

  • Cope HG, Williams MA (1981) Secretory granule formation in the rabbit parotid gland after isoprenaline induced secretion, stereological reconstructions of granule populations. Anat Rec 199:377–387

    Google Scholar 

  • Crine P, Gianoulakis C, Seideh NG, Gossard F, Pezalla PD, Lis M, Chrétien M (1978) Biosynthesis of β-endorphin from β-lipotropin and a larger molecular weight precursor in rat pars intermedia. Proc Natl Acad Sci USA 75:4719–4722

    Google Scholar 

  • Elmalek M, Hammel I (1987) Morphometric evidence that the maturation of the eosinophil granules is independent of volume change. J Submicrose Cytol 19:265–268

    Google Scholar 

  • Farquhar MG, Palade GE (1981) The Golgi apparatus (complex) (1954–1981) — from artifact to center stage. J Cell Biol 91:77s-103s

    Google Scholar 

  • Glembotski CC (1981) Subcellular fractionation studies on the post-translational processing of pro-adrenocorticotropic hormone/endorpin in the rat intermediate pituitary. J Biol Chem 256:7433–7439

    Google Scholar 

  • Glembotski CC (1982) Acetylation of γ-melanotropin and β-endorphin in the rat intermediate pituitary. J Biol Chem 257:10493–10500

    Google Scholar 

  • Hammel I, Lagunoff D, Bauza M, Chi E (1983) Periodic, multimodal distribution of granule volumes in mast cells. Cell Tissue Res 228:51–59

    Google Scholar 

  • Hammel I, Dvorak AM, Peters SP, Schulman ES, Dvorak HF, Lichtenstein LM, Galli SJ (1985) Differences in the volume distribution of human lung mast cell granules and lipid bodies: Evidence that the size of these organelles is regulated by distinct mechanism. J Cell Biol 100:1488–1492

    Google Scholar 

  • Hammel I, Dvorak AM, Galli SJ (1987) Defective cytoplasmic granule formation. I. Abnormalities affecting tissue mast cells and pancreatic acinar cells of beige mice. Lab Invest 56:321–328

    Google Scholar 

  • Knecht E, Martinez-Ramon A, Grisolia S (1986) Electron microscopic localization of glutamate dehydrogenase in rat liver mitochondria by an immunogold procedure and monoclonal and polyclonal antibodies. J Histochem Cytochem 34:913–922

    Google Scholar 

  • Loh YP, Gainer H (1982) Characterization of pro-opiocortin converting activity in purified secretory granules from rat pituitary neurointermediate lobe. Proc Natl Acad Sci USA 79:108–112

    Google Scholar 

  • Loh YP, Gritsch HA, Chang T-L (1982) Pro-opiocortin processing in the pituitary: a model for neuropeptide biosynthesis. Peptides 3:397–404

    Google Scholar 

  • Mains RE, Eipper BA, Ling N (1977) Common precursor to corticotropins and endorphins. Proc Natl Acad Sci USA 74:3014–3018

    Google Scholar 

  • Mains RE, Eipper BA, Glembotski CC, Dores RM (1983) Strategies for the biosynthesis of bioactive peptides. Trends Neurosci June:229–235

  • Mroz MA, Lechene C (1986) Pancreatic zymogen granules differ markedly in protein composition. Science 232:871–873

    Google Scholar 

  • Orci L, Ravazola M, Amherdt M, Madsen O, Vassali J-D, Perrelet A (1985) Direct identification of prohormone conversion site to insulin-secreting cells. Cell 42:671–681

    Google Scholar 

  • Scott AP, Lowry PJ, Ratcliffe JG, Rees LH Landon J (1974) Corticotrophin-like peptides in the rat pituitary. J Endocrinol 61:355–367

    Google Scholar 

  • Smith RE, Farquhar MG (1966) Lysosome function in the regulation of the secretory process in cells of the anterior pituitary gland. J Cell Biol 31:319–347

    Google Scholar 

  • Sokal RR, Rohlf FJ (1969) Biometry. Freeman, San Francisco

    Google Scholar 

  • Stoekel ME, Schimchowitsch S, Garaud JC, Schimitt G, Vaudry H, Klein MJ, Porte A (1985) Immunocytochemical evidence for intragranular processing of pro-opiomelanocortin in the melanotropic cells of the rabbit. Cell Tissue Res 242:365–370

    Google Scholar 

  • Tooze J, Hollinshead M, Frank R, Buzke B (1987) An antibody specific for an endoproteolitic cleavage site provides evidence that pro-opiomelanocortin is packaged into secretory granules in AtT20 cells before its cleavage. J Cell Biol 105:155–162

    Google Scholar 

  • Varndell IM, Tapia FJ, Probert L, Buchman AMJ, Gu J, De May J, Bloom SR, Polak JM (1982) Immunogold staining procedure for the localization of regulatory peptides. Peptides 3:259–272

    Google Scholar 

  • Varndell IM, Sikri KL, Hennessy RJ, Kalina M, Goodman RH, Benoit R, Diani AR, Polak JM (1986) Mammalian somatostatin-containing D cells exhibits rat somatostatin cryptic peptide (RSCP) immunoreactivity: Electron microscopical study. Cell Tissue Res 246:196–204

    Google Scholar 

  • Williams MA (1977) Quantitative methods in biology North-Holland New York, pp 5–84

    Google Scholar 

  • Williams MA, Cope HG (1981) Membrane dynamics in the parotid acinar cell during regranulation: A stereological study following isoprenaline-induced secretion. Anat Rec 199:389–401

    Google Scholar 

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Kalina, M., Elmalek, M. & Hammel, I. Intragranular processing of pro-opiomelanocortin in the intermediate cells of the rat pituitary glands. Histochemistry 89, 193–198 (1988). https://doi.org/10.1007/BF00489924

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