Skip to main content
SpringerLink
Log in

Protein kinase C isoenzymes in mouse Harderian gland. Differential expression of the α- and ε-isoforms during pregnancy

Protein kinase C-OC

  • Original Paper
  • Published:
Histochemistry and Cell Biology Aims and scope Submit manuscript

Abstract

Protein kinase C (PKC) is known to be involved in the regulation of exocytosis in different cell lines and tissues. Experiments were designed to determine whether the Harderian gland of CD-1 mouse produces PKC isoenzymes and whether the expression of the isoforms changes during pregnancy. The presence of the isoenzymes was assessed by immunoblotting experiments using extract of total Harderian gland and polyclonal antisera specific for nine different PKC isoforms. Antisera giving a positive staining on Western blots were subsequently used for immunohistochemical investigation using a secondary antibody conjugated to alkaline phosphatase. Immunoblotting experiments revealed that the Harderian gland from female mouse expresses PKC isoforms-α,-ε,-ζ and-η. These isoforms were also detected in the Harderian gland from 13-day pregnant mouse; however, striking quantitative changes were seen concerning the α- and ε-isoforms. The 80-kDa native form of PKC-α almost doubled in the pregnant mouse in comparison with normal female mouse whereas the amount of 50-kDa catalytic domain did not change. Protein kinase C-ε appeared as a 92- to 93-kDa form and a 67-kDa form. While the 92- to 93-kDa protein was expressed to a similar extent in both types of mouse, the 67-kDa form was more abundant inthe Harderian gland from normal female mouse. These data were corroborated by immunohistochemical experiments and showing a diffuse and granular staining of the adenomeres. These observations demonstrate for the first time (to our knowledge) that the mouse Harderian gland produces several PKC isoenzymes that could be involved in the regulation of exocytosis and/or other functions. Moreover, the expression of the α- and ε-isoforms could be regulated by sexual hormones, as suggested by the differential abundance of these two proteins in the gland of pregnant mouse compared with normal female mouse.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Akagi N, Morimoto T, Watanabe M (1987) The Harderian gland and the porphyrins. Bull Osaka Med Sch 33:97–115

    PubMed  Google Scholar 

  • Akita Y, Ohno S, Yajima Y, Konno Y, Saido TC, Mizuno K, Chida K, Osada S, Kuroki T, Kawashima S, Suzuki K (1994) Over-production of a Ca2+-independent protein kinase C isozyme, nPKCε, increases the secretion of prolactin from thyrotropin-releasing hormone-stimulated rat pituitary GH4C1 cells. J Biol Chem 269:4653–4660

    PubMed  Google Scholar 

  • Ben-Shlomo H, Sigmund O, Stabel S, Reiss N, Naor Z (1991) Preferential release of catecholamine from permeabilized PC12 cells by α- and β-type protein kinase C subspecies. Biochem J 280:65–69

    PubMed  Google Scholar 

  • Berti L, Mosthaf L, Kroder G, Kellerer M, Tippmer S, Mushack J, Seffer E, Seedorf K, Haring H (1994) Glucose-induced translocation of protein kinase C isoforms in rat-1 fibroblasts is paralleled by inhibition of the insulin receptor tyrosine kinase. J Biol Chem 269:3381–3386

    PubMed  Google Scholar 

  • Brooksbank BWL, wilson DAA, Clough G (1973) The in vivo uptake of3H-androsta-4, 16-dien-3-one in tissues of the adult male rat. J Endocrinol 57:1–11

    PubMed  Google Scholar 

  • Cohn SA (1954) Histochemical observations of the Harderian gland of the albino mouse. J Histochem Cytochem 3:342–353

    Google Scholar 

  • Connor K, Clegg RA (1993) Isoenzymes of protein kinase C in rat mammary tissues: changes in properties and relative amounts during pregnancy and lactation. Biochem J 291:817–824

    PubMed  Google Scholar 

  • Dekker LV, Parker PJ (1994) Protein kinase C: a question of specificity. Trends Biochem Sci 19: 73–77

    Article  PubMed  Google Scholar 

  • Gesase AP, Satoh Y, Ono K, Habara Y, Kanno T (1994) Exocrine mechanism in Harderian gland, with special reference to involvement of G-proteins and extracellular and intracellular Ca2+. Proceedings of the International Symposium on the Cell Biology and Physiology of the Harderian Gland, Sorrento, Italy, June 2–6, 1994, XII (Abstr)

  • Gschwendt M, Leibersperger H, Rincke G, Marks F (1991) Immunological demonstration of εPKC. Murine tissue distribution, ontogeny, cellular localization and translocation. FEBS Lett 290:115–118

    Article  PubMed  Google Scholar 

  • Hug H, Sarre TF (1993) Protein kinase C isoenzymes: divergence in signal transduction? Biochem J 291:329–343

    PubMed  Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the bacteriophage T4. Nature 227:680–685

    PubMed  Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275

    PubMed  Google Scholar 

  • Martelli AM, Neri LM, Gilmour RS, Barker PJ, Huskisson NS, Manzoli FA, Cocco L (1991a) Temporal changes in intracellular distribution of protein kinase C in Swiss 3T3 cells during mitogenic stimulation with insulin-like growth factor I and bombesin: translocation to the nucleus follows rapid changes in nuclear polyphosphoinositides. Biochem Biophys Res Commun 177:480–487

    Article  PubMed  Google Scholar 

  • Martelli AM, Carini C, Marmiroli S, Mazzoni M, Barker PJ, Gilmour RS, Capitani S (1991b) Nuclear protein kinases in rat liver: evidence for increased histone H1 phosphorylating activity during liver regeneration. Exp Cell Res 195:255–262

    PubMed  Google Scholar 

  • Morgan A, Burgoyne RD (1992) Exo1 and Exo2 proteins stimulate calcium-dependent exocytosis in permeabilized adrenal chromaffin cells. Nature 355:833–836

    Article  PubMed  Google Scholar 

  • Neri LM, Billi AM, Manzoli L, Rubbini S, Gilmour RS, Cocco L, Martelli AM (1994) Selective nuclear translocation of protein kinase C α in Swiss 3T3 cells treated with IGF-I, PDGF and EGF. FEBS Lett 347:63–68

    PubMed  Google Scholar 

  • Nishizaki T, Walent JH, Kowalchyk JA, Martin TF (1992) A key role for a 145-kDa cytosolic protein in the stimulation of Ca(2+)-dependent secretion by protein kinase C. J Biol Chem 267:23972–23981

    PubMed  Google Scholar 

  • Nishizuka Y (1986) Studies and perspectives of protein kinase C. Science 233:305–312

    PubMed  Google Scholar 

  • Nishizuka Y (1989) Studies and prospectives of the protein kinase C family for cellular regulation. Cancer 63:1892–1903

    PubMed  Google Scholar 

  • Otsuru O, Otsuka H, Kasama T, Seyama Y, Sakai T, Yohro T (1983) The characterization of 2,3-alkanediol diacyl esters obtained from the Harderian glands of Mongolian gerbil (Meriones unguiculatus). J Biochem 94:2049–2054

    PubMed  Google Scholar 

  • Ozawa K, Szallasi Z, Kazanietz MG, Blumberg PM, Mischak H, Mushinski JF, Beaven MA (1993) Ca(2+)-dependent and Ca(2+)-independent isozymes of protein kinase C mediate exocytosis in antigen-stimulated rat basophili RBL-2H3 cells. Reconstitution of secretory responses with Ca(2+) and purified isozymes in washed permeabilized cells. J Biol Chem 268:2280–2283

    PubMed  Google Scholar 

  • Park SH, Kumagai S, Seyama Y (1994) Porphyrin localization in mouse Harderian gland cells as analyzed by interactive laser cytometer. Proceedings of the International Symposium on the Cell Biology and Physiology of the Harderian Gland, Sorrento, Italy, June 2–6, 1994, XI (Abstr)

  • Sakai T (1992) Comparative anatomy of mammalian Harderian glands. In: Webb SM, Hoffman RA, Puig-Domingo ML, Reiter RJ (eds) Harderian gland. Porphyrin metabolism, behavioral and endocrine effects. Springer, Verlag, Berlin Heidelberg, pp 7–23

    Google Scholar 

  • Sakai T, Yohro T (1981) A histological study of the Harderian gland of Mongolian gerbils,Meriones meridianus. Anat Rec 200:259–270

    Article  PubMed  Google Scholar 

  • Shirama K, Hokano M (1992) Harderian glands and their development in laboratory rats and mice. In: Webb SM, Hoffman RA, Puig-Domingo ML, Reiter RJ (eds) Harderian gland. Porphyrin metabolism, behavioral and endocrine effects. Springer, Berlin Heidelberg, pp 25–51

    Google Scholar 

  • Shirama K, Furuya T, Takeo Y, Shimizu K, Maekawa K (1981) Influences of some endocrine glands and of hormone replacement on the porphyrins of the Harderian glands of mice. J Endocrinol 91:305–311

    PubMed  Google Scholar 

  • Simboli-Campbell M, Gagnon AM, Franks DJ, Welsh JE (1994) 1,25-dihydroxyvitamin D3 translocates protein kinase Cβ to nucleus and enhances plasma membrane association of protein kinase Cα in renal epithelial cells. J Biol Chem 269:3357–3364

    Google Scholar 

  • Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets. Proc Natl Acad Sci USA 76:4350–4354

    PubMed  Google Scholar 

  • Turner NA, Rumsby MG, Walker JH, McMorris FA, Ball SG, Vaughan PFT (1994) A role for protein kinase C subtypes α and ε in phorbol-ester-enhanced K+- and carbachol-evoked noradrenaline release from the human neuroblastoma SHSY5Y. Biochem J 297:407–413

    PubMed  Google Scholar 

  • Ulrich R, Yuwiler A, Geller E, Wetterberg L (1974) Effects of sex hormones and environmental lighting on rat Harderian gland porphyrin. J Endocrinol 63:99–102

    PubMed  Google Scholar 

  • Wetsel WC, Khan WA, Merchentaler I, Rivera H, Halpern AE, Phung HM, Negro-Vilar A, Hannun YA (1992) Tissue and cellular distribution of the extended family of protein kinase C isoenzymes. J Cell Biol 117:121–133

    Article  PubMed  Google Scholar 

  • Wooding FBP (1980) Lipid droplet secretion by the rabbit Harderian gland. J Ultrastruct Res 71:68–78

    Article  PubMed  Google Scholar 

  • Zini N, Santi S, Ognibene A, Bavelloni A, Neri LM, Valmori A, Mariani E, Negri C, Astaldi-Ricotti GCB, Maraldi NM (1994) Discrete localization of different DNA topoisomerases in HeLa and K562 cell nuclei and subnuclear fractions. Exp Cell Res 210:336–348

    Article  PubMed  Google Scholar 

  • Zoukhri D, Sergheraert C, Rossignol B (1993) Phorbol ester-stimulated exocytosis in lacrimal gland: PKC might not be the sole effector. Am J Physiol 264:1045–1050

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Dipartimento di Morfologia Umana Normale, Università di Trieste, via Manzoni 16, I-34138, Trieste, Italy

    Vittorio Grill, Alberto M. Martelli, Renato Bareggi, Marisa Basa, Marina Zweyer & Paola Narducci

  2. Istituto di Citomorfologia Normale e Patologica del C.N.R., c/o I.O.R., via di Barbiano 1/10, I-40136, Bologna, Italy

    Spartaco Santi

  3. Istituto di Anatomia Umana Normale, via Irnerio 48, I-40126, Bologna, Italy

    Lucio Cocco

Authors
  1. Vittorio Grill
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Alberto M. Martelli
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Renato Bareggi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Spartaco Santi
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Marisa Basa
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Marina Zweyer
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Lucio Cocco
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Paola Narducci
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Grill, V., Martelli, A.M., Bareggi, R. et al. Protein kinase C isoenzymes in mouse Harderian gland. Differential expression of the α- and ε-isoforms during pregnancy. Histochem Cell Biol 103, 255–262 (1995). https://doi.org/10.1007/BF01457409

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01457409

Keywords

Search

Navigation