title: Apocrine secretion in Drosophila salivary glands: subcellular origin, dynamics, and identification of secretory proteins.
abstract: In contrast to the well defined mechanism of merocrine exocytosis, the mechanism of apocrine secretion, which was first described over 180 years ago, remains relatively uncharacterized. We identified apocrine secretory activity in the late prepupal salivary glands of Drosophila melanogaster just prior to the execution of programmed cell death (PCD). The excellent genetic tools available in Drosophila provide an opportunity to dissect for the first time the molecular and mechanistic aspects of this process. A prerequisite for such an analysis is to have pivotal immunohistochemical, ultrastructural, biochemical and proteomic data that fully characterize the process. Here we present data showing that the Drosophila salivary glands release all kinds of cellular proteins by an apocrine mechanism including cytoskeletal, cytosolic, mitochondrial, nuclear and nucleolar components. Surprisingly, the apocrine release of these proteins displays a temporal pattern with the sequential release of some proteins (e.g. transcription factor BR-C, tumor suppressor p127, cytoskeletal β-tubulin, non-muscle myosin) earlier than others (e.g. filamentous actin, nuclear lamin, mitochondrial pyruvate dehydrogenase). Although the apocrine release of proteins takes place just prior to the execution of an apoptotic program, the nuclear DNA is never released. Western blotting indicates that the secreted proteins remain undegraded in the lumen. Following apocrine secretion, the salivary gland cells remain quite vital, as they retain highly active transcriptional and protein synthetic activity.
authors.affiliation: Laboratory of Developmental Genetics, Institute of Experimental Endocrinology, Slovak Academy of Sciences, Bratislava, Slovakia.
authors.forename: Robert
authors.initials: R
authors.lastname: Farkaš
authors.affiliation: Laboratory of Developmental Genetics, Institute of Experimental Endocrinology, Slovak Academy of Sciences, Bratislava, Slovakia,; Department of Genetics, Comenius University, Bratislava, Slovakia.
authors.forename: Zuzana
authors.initials: Z
authors.lastname: Ďatková
authors.affiliation: Laboratory of Developmental Genetics, Institute of Experimental Endocrinology, Slovak Academy of Sciences, Bratislava, Slovakia,; Department of Genetics, Comenius University, Bratislava, Slovakia.
authors.forename: Lucia
authors.initials: L
authors.lastname: Mentelová
authors.affiliation: Department of Anatomy and Cell Biology, Lorand Eötvös University, Budapest, Hungary.
authors.forename: Péter
authors.initials: P
authors.lastname: Löw
authors.affiliation: Laboratory of Developmental Genetics, Institute of Experimental Endocrinology, Slovak Academy of Sciences, Bratislava, Slovakia.
authors.forename: Denisa
authors.initials: D
authors.lastname: Beňová-Liszeková
authors.affiliation: Laboratory of Developmental Genetics, Institute of Experimental Endocrinology, Slovak Academy of Sciences, Bratislava, Slovakia.
authors.forename: Milan
authors.initials: M
authors.lastname: Beňo
authors.affiliation: Department of Anatomy and Cell Biology, Lorand Eötvös University, Budapest, Hungary.
authors.forename: Miklós
authors.initials: M
authors.lastname: Sass
authors.affiliation: Institute of Molecular Pathology, Faculty of Military Health Sciences, University of Defence, Hradec Králové, Czech Republic.
authors.forename: Pavel
authors.initials: P
authors.lastname: Řehulka
authors.affiliation: 1st Department of Internal Medicine - Cardioangiology, Faculty of Medicine in Hradec Králové, Charles University in Prague, Hradec Králové, Czech Republic.
authors.forename: Helena
authors.initials: H
authors.lastname: Řehulková
authors.affiliation: Institute of Cellular Biology and Pathology, 1st Faculty of Medicine, Charles University in Prague, Prague, Czech Republic.
authors.forename: Otakar
authors.initials: O
authors.lastname: Raška
authors.affiliation: Institute of Cellular Biology and Pathology, 1st Faculty of Medicine, Charles University in Prague, Prague, Czech Republic.
authors.forename: Lubomír
authors.initials: L
authors.lastname: Kováčik
authors.affiliation: Institute of Cellular Biology and Pathology, 1st Faculty of Medicine, Charles University in Prague, Prague, Czech Republic.
authors.forename: Jana
authors.initials: J
authors.lastname: Šmigová
authors.affiliation: Institute of Cellular Biology and Pathology, 1st Faculty of Medicine, Charles University in Prague, Prague, Czech Republic.
authors.forename: Ivan
authors.initials: I
authors.lastname: Raška
authors.affiliation: Institute of Cellular Biology and Pathology, 1st Faculty of Medicine, Charles University in Prague, Prague, Czech Republic; Deutsches Krebsforschungszentrum, Heidelberg, Germany.
authors.forename: Bernard M
authors.initials: BM
authors.lastname: Mechler
mesh_terms: D000818:Animals; D001050:Apocrine Glands / Q000378:metabolism* / Q000648:ultrastructure; D004247:DNA / Q000378:metabolism; D029721:Drosophila Proteins / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000378:metabolism; D007814:Larva / Q000254:growth & development / Q000378:metabolism; D014176:Protein Biosynthesis; D011679:Pupa / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012469:Salivary Glands / Q000378:metabolism* / Q000648:ultrastructure; D012471:Salivary Proteins and Peptides / Q000378:metabolism*; D013347:Subcellular Fractions / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D029721:Drosophila Proteins; D005456:Fluorescent Dyes; D011993:Recombinant Fusion Proteins; D012471:Salivary Proteins and Peptides; D004247:DNA
references.citation: Cell. 1990 Apr 6;61(1):85-99
references.pmid: 2107982
references.citation: Int J Biochem Cell Biol. 2010 Feb;42(2):181-5
references.pmid: 19944183
references.citation: J Cell Biol. 2010 Feb 8;188(3):401-13
references.pmid: 20142423
references.citation: Int Rev Cytol. 1984;87:253-329
references.pmid: 6370891
references.citation: J Mass Spectrom. 2010 Jan;45(1):104-11
references.pmid: 19927305
references.citation: J Cell Biol. 2009 Mar 9;184(5):751-64
references.pmid: 19255244
references.citation: Am Surg. 2008 Dec;74(12):1177-81
references.pmid: 19097532
references.citation: Laryngoscope. 1976 Aug;86(8):1164-76
references.pmid: 950858
references.citation: BMC Med. 2013 Apr 04;11:93
references.pmid: 23556533
references.citation: Acta Anat (Basel). 1992;143(1):7-13
references.pmid: 1350161
references.citation: J Proteome Res. 2009 Nov;8(11):4889-905
references.pmid: 19705875
references.citation: Z Zellforsch Mikrosk Anat. 1972;127(1):50-86
references.pmid: 4336139
references.citation: J Cell Biol. 2010 Feb 8;188(3):415-28
references.pmid: 20142424
references.citation: Anat Rec. 1998 Nov;252(3):403-12
references.pmid: 9811218
references.citation: Rhinology. 1986 Mar;24(1):57-60
references.pmid: 3704466
references.citation: Exp Dermatol. 2011 Oct;20(10):841-2
references.pmid: 21824197
references.citation: Ital J Anat Embryol. 2007 Apr-Jun;112(2):117-29
references.pmid: 17687876
references.citation: Histol Histopathol. 2003 Apr;18(2):597-608
references.pmid: 12647810
references.citation: Anat Embryol (Berl). 2000 Jan;201(1):51-61
references.pmid: 10603093
references.citation: Cell Tissue Res. 1996 Sep;285(3):501-7
references.pmid: 8772164
references.citation: Anat Embryol (Berl). 1995 Oct;192(4):319-28
references.pmid: 8554165
references.citation: Results Probl Cell Differ. 1972;4:101-51
references.pmid: 4198830
references.citation: Eye (Lond). 2010 Aug;24(8):1396-402
references.pmid: 20150925
references.citation: JAAPA. 2008 Nov;21(11):23-5
references.pmid: 19105543
references.citation: Wilhelm Roux Arch Entwickl Mech Org. 1972 Sep;169(3):216-238
references.pmid: 28304626
references.citation: FEBS Lett. 2009 Dec 3;583(23):3804-10
references.pmid: 19887068
references.citation: Nature. 1970 Aug 15;227(5259):680-5
references.pmid: 5432063
references.citation: Dev Biol. 1976 Dec;54(2):256-63
references.pmid: 825404
references.citation: J Biol Chem. 1969 Aug 25;244(16):4406-12
references.pmid: 5806584
references.citation: Ann Diagn Pathol. 2010 Feb;14(1):1-7
references.pmid: 20123450
references.citation: Bioessays. 1996 Jan;18(1):47-54
references.pmid: 8593163
references.citation: Nat Struct Mol Biol. 2008 Jul;15(7):665-74
references.pmid: 18618940
references.citation: Nat Rev Mol Cell Biol. 2005 Feb;6(2):181-7
references.pmid: 15688003
references.citation: J Electron Microsc (Tokyo). 1966;15(2):94-5
references.pmid: 5965757
references.citation: Adv Exp Med Biol. 2011;716:107-22
references.pmid: 21713654
references.citation: J Submicrosc Cytol. 1981 Jul;13(3):473-8
references.pmid: 6174736
references.citation: J Oral Maxillofac Surg. 2010 Feb;68(2):451-5
references.pmid: 20116722
references.citation: Microsc Res Tech. 1996 Jun 1;34(2):104-10
references.pmid: 8722703
references.citation: Pediatrics. 2005 Feb;115(2):e239-41
references.pmid: 15629957
references.citation: J Invest Dermatol. 1975 Jan;64(1):38-41
references.pmid: 1110304
references.citation: J Morphol. 2000 May;244(2):137-42
references.pmid: 10761051
references.citation: Cell. 2007 Jan 12;128(1):183-95
references.pmid: 17218264
references.citation: Cell Tissue Res. 1999 Mar;295(3):495-504
references.pmid: 10022969
references.citation: Insect Biochem Mol Biol. 2002 Feb;32(2):113-20
references.pmid: 11755052
references.citation: Br J Dermatol. 2013 Feb;168(2):243-52
references.pmid: 23106519
references.citation: Biochimie. 2001 Mar-Apr;83(3-4):301-10
references.pmid: 11295490
references.citation: Histopathology. 2008 Jan;52(1):3-10
references.pmid: 18171412
references.citation: J Am Acad Dermatol. 2010 Feb;62(2):205-17
references.pmid: 20115947
references.citation: Z Mikrosk Anat Forsch. 1982;96(1):130-7
references.pmid: 7102032
references.citation: Am Fam Physician. 2005 Oct 15;72(8):1547-52
references.pmid: 16273821
references.citation: Genes Dev. 1989 Sep;3(9):1288-300
references.pmid: 2558050
references.citation: J Invest Dermatol. 2003 Feb;120(2):175-81
references.pmid: 12542519
references.citation: In Vitro Cell Dev Biol Anim. 1998 Nov-Dec;34(10):813-23
references.pmid: 9870531
references.citation: Clin Lab Med. 2005 Dec;25(4):761-75, vii
references.pmid: 16308090
references.citation: Anal Biochem. 1980 Jul 1;105(2):361-3
references.pmid: 6161559
references.citation: Gene. 1991 Dec 30;109(2):265-7
references.pmid: 1765271
references.citation: Chromosoma. 1989 Aug;98(2):81-5
references.pmid: 2476281
references.citation: J Histochem Cytochem. 1998 Apr;46(4):505-11
references.pmid: 9524196
references.citation: Dev Biol. 1976 Jan;48(1):191-5
references.pmid: 812742
references.citation: Dev Biol. 1977 Jul 15;58(2):339-55
references.pmid: 407116
references.citation: J Electron Microsc Tech. 1989 Feb;11(2):117-25
references.pmid: 2709130
references.citation: Genes Dev. 1990 Feb;4(2):204-19
references.pmid: 2110921
references.citation: J Mol Biol. 1974 Apr 15;84(3):389-98
references.pmid: 4219221
references.citation: Trends Genet. 1996 Aug;12(8):306-10
references.pmid: 8783940
references.citation: Wilehm Roux Arch Dev Biol. 1981 Nov;190(6):351-357
references.pmid: 28305294
references.citation: Cell Death Differ. 2000 Jan;7(1):89-101
references.pmid: 10713724
references.citation: Eur Biophys J. 2011 Sep;40(9):1013-27
references.pmid: 21833780
references.citation: Arch Biochem Biophys. 2003 Jul 1;415(1):1-5
references.pmid: 12801506
references.citation: J Anat. 1994 Aug;185 ( Pt 1):1-49
references.pmid: 7559104
references.citation: Cell Mol Life Sci. 2004 Jul;61(13):1546-61
references.pmid: 15224180
references.citation: Nat Protoc. 2007;2(8):1896-906
references.pmid: 17703201
references.citation: Ann N Y Acad Sci. 2004 Apr;1014:170-8
references.pmid: 15153432
references.citation: Biol Reprod. 1995 Jan;52(1):50-62
references.pmid: 7711183
references.citation: Science. 2009 Jan 23;323(5913):474-7
references.pmid: 19164740
references.citation: J Cutan Pathol. 2008 Jul;35(7):701-3
references.pmid: 18582247
references.citation: Cell Mol Life Sci. 2013 Jun;70(12):2099-121
references.pmid: 22986507
references.citation: Neuroreport. 2004 Jan 19;15(1):79-81
references.pmid: 15106835
references.citation: Eur J Biochem. 1975 Aug 15;56(2):335-41
references.pmid: 1175627
references.citation: Annu Rev Biochem. 1999;68:1015-68
references.pmid: 10872471
references.citation: J Electron Microsc (Tokyo). 1968;17(2):158-9
references.pmid: 4177281
references.citation: Invest Ophthalmol Vis Sci. 2005 Mar;46(3):863-76
references.pmid: 15728542
references.citation: Genetics. 2009 Feb;181(2):543-65
references.pmid: 19064711
references.citation: Cell Biochem Biophys. 2006;45(1):111-23
references.pmid: 16679567
references.citation: Nature. 2008 May 22;453(7194):481-8
references.pmid: 18497817
references.citation: Nucleic Acids Res. 1983 Apr 25;11(8):2411-25
references.pmid: 6304652
references.citation: Development. 1997 Nov;124(22):4673-83
references.pmid: 9409683
references.citation: Cell Mol Life Sci. 2008 Sep;65(18):2814-32
references.pmid: 18726177
references.citation: Annu Rev Neurosci. 2004;27:509-47
references.pmid: 15217342
references.citation: Mod Pathol. 2006 Feb;19 Suppl 2:S93-S126
references.pmid: 16446719
references.citation: FEBS Lett. 2009 Sep 3;583(17):2701-9
references.pmid: 19631211
references.citation: Cell Calcium. 2005 Sep-Oct;38(3-4):343-53
references.pmid: 16099500
references.citation: Microsc Res Tech. 2007 Dec;70(12):1022-7
references.pmid: 17661387
references.citation: Physiol Rev. 2012 Oct;92(4):1915-64
references.pmid: 23073634
references.citation: Cell Death Differ. 2003 Sep;10(9):940-5
references.pmid: 12934068
references.citation: Nat Protoc. 2006;1(6):2856-60
references.pmid: 17406544
references.citation: J Anat. 1978 Oct;127(Pt 2):291-8
references.pmid: 721691
references.citation: Nature. 2012 Oct 11;490(7419):201-7
references.pmid: 23060190
references.citation: J Anat. 1959 Apr;93(2):226-32
references.pmid: 13641121
references.citation: Nature. 2011 Mar 24;471(7339):473-9
references.pmid: 21179090
references.citation: Nat Genet. 2007 Jun;39(6):715-20
references.pmid: 17534367
references.citation: Curr Opin Cell Biol. 2009 Aug;21(4):537-42
references.pmid: 19473826
references.citation: J Neurosci. 2007 Apr 18;27(16):4231-2
references.pmid: 17442806
references.citation: J Am Acad Nurse Pract. 2007 May;19(5):228-34
references.pmid: 17489955
references.citation: Cell Mol Life Sci. 2009 Sep;66(17):2819-33
references.pmid: 19468686
references.citation: Graefes Arch Clin Exp Ophthalmol. 2000 Sep;238(9):738-45
references.pmid: 11045341
references.citation: Ann Anat. 1999 Sep;181(5):437-46
references.pmid: 10560009
references.citation: Mol Cell. 2000 Mar;5(3):445-55
references.pmid: 10882130
references.citation: Histochemistry. 1990;93(5):525-30
references.pmid: 1970553
references.citation: Cold Spring Harb Symp Quant Biol. 1974;38:655-62
references.pmid: 4208797
references.citation: Nat Rev Mol Cell Biol. 2011 Aug 10;12(9):581-93
references.pmid: 21829222
references.citation: Nature. 1983 Jul 21-27;304(5923):234-41
references.pmid: 6408489
references.citation: Eur J Cell Biol. 1994 Oct;65(1):49-59
references.pmid: 7889995
references.citation: Stain Technol. 1983 Nov;58(6):347-51
references.pmid: 6679126
references.citation: J Invest Dermatol. 1954 Feb;22(2):143-56
references.pmid: 13130897
references.citation: Biotech Histochem. 1993 Jul;68(4):237-9
references.pmid: 8218577
references.citation: Genes Dev. 1994 May 15;8(10):1160-73
references.pmid: 7926721
references.citation: Dev Biol. 1976 Dec;54(2):264-75
references.pmid: 825405
references.citation: Anat Rec. 1979 Feb;193(2):269-91
references.pmid: 426299
references.citation: CNS Neurol Disord Drug Targets. 2006 Feb;5(1):57-77
references.pmid: 16613554
references.citation: Microsc Res Tech. 1993 Dec 15;26(6):489-95
references.pmid: 8305726
references.citation: Science. 2009 Jan 23;323(5913):516-21
references.pmid: 19164751
references.citation: J Electron Microsc (Tokyo). 1964;13(3):139-44
references.pmid: 4862357
references.citation: J Cell Biol. 1963 Apr;17:208-12
references.pmid: 13986422
references.citation: Cell. 2006 Aug 11;126(3):597-609
references.pmid: 16901790
references.citation: J Biophys Biochem Cytol. 1958 Jul 25;4(4):475-8
references.pmid: 13563554
references.citation: Proc Natl Acad Sci U S A. 1975 Nov;72(11):4550-4
references.pmid: 812097
references.citation: Annu Rev Entomol. 2012;57:187-204
references.pmid: 21910636
references.citation: Wilehm Roux Arch Dev Biol. 1975 Mar;178(1):15-28
references.pmid: 28305064