The expression patterns and the dynamic changes in content of both annexin I and annexin II in the rat pancreatic islets during postnatal development were investigated by both western blot analysis and immunohistochemistry. Immunohistochemical methods clearly demonstrated the presence of annexins I and II exclusively in pancreatic islets, while exocrine tissues were not stained by anti-annexin antibodies. Pancreatic islets were diffusely stained with no specific differences in distribution between different cell types. The expression of annexin I in pancreatic islets gradually increased with postnatal development. A developmental study of annexins I and II by western blot analysis essentially supported the results obtained by immunohistochemistry. In addition, the increasing expression of two protein tyrosine kinases, epidermal growth factor-receptor/kinase and pp60src, which phosphorylate annexin I and annexin II, respectively, and of protein kinase C, which phosphorylates both proteins, was also shown during postnatal development in rat pancreatic islets. Thus, a relationship between the expression of annexins I and II and the maturation of islet cell function is suggested.

REFERENCES

Ali
S. M.
,
Geisow
M. J.
,
Burgoyne
R. D.
(
1989
).
A role for calpactin in calcium dependent exocytosis in adrenal chromaffin cells.
Nature
340
,
313
315
Asplund
K.
(
1973
).
Effect of glucose on insulin biosynthesis in fetal and newborn rats.
Horm. Metab. Res
5
,
410
415
Blanquet
P. R.
,
Paillard
S.
,
Courtosin
Y.
(
1988
).
Influence of fibroblast growth factor on phosphorylation and activity of a 34 kDa lipocortin-like protein in bovine epithelial lens cells.
FEBS Lett
229
,
183
187
Borregaard
N.
,
Kjeldsen
L.
,
Lollike
K.
,
Sengelov
H.
(
1992
).
Ca2+-dependent translocation of cytosolic proteins to isolated granule subpopulations and plasma membrane from human neutrophils.
FEBS Lett
304
,
195
197
Bradford
M. M.
(
1976
).
A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
Anal. Biochem
72
,
248
254
Brugge
J. S.
(
1986
).
The p35/p36 substrates of protein tyrosine kinases as inhibitors of phospholipase A2.
Cell
46
,
149
150
Burgoyne
R. D.
(
1988
).
Calpactin in exocytosis?.
Nature
331
,
20
–.
Burgoyne
R. D.
,
Cambray-Deakin
M. A.
,
Norman
K. M.
(
1989
).
Developmental regulation of tyrosine kinase substrate p36 (calpactin heavy chain) in rat cerebellum.
J. Mol. Neurosci
1
,
47
54
Burgoyne
R. D.
,
Geisow
M. J.
(
1989
).
The annexin family of calcium-binding proteins.
Cell Calcium
10
,
1
10
Calle
R.
,
Ganesan
S.
,
Smallwood
J. I.
,
Rasmussen
H.
(
1992
).
Glucose-induced phosphorylation of myristoylated alanine-rich C kinase substrate (MARCKS) in isolated rat pancreatic islets.
J. Biol. Chem
267
,
18723
18727
Chang
K. S.
,
Wang
G.
,
Freireich
E. J.
,
Daly
M.
,
Naylor
S. L.
,
Trujillo
J. M.
,
Stass
S. A.
(
1992
).
Specific expression of the annexin XIII gene in acute promyelocytic leukemia.
Blood
79
,
1802
1810
Clark
D. M.
,
Moss
S. E.
,
Wright
N. A.
,
Crumpton
M. J.
(
1991
).
Expression of annexin VI (p68, 67 kDa-calelectrin) in normal human tissues: evidence for developmental regulation in B-and T-lymphocytes.
Histochemistry
96
,
405
412
Colca
J. R.
,
Brooks
C. L.
,
Landt
M.
,
McDaniel
M. L.
(
1983
).
Correlation of Ca2+-and calmodulin-dependent protein kinase activity with secretion of insulin from islets of Langerhans.
J. Biol. Chem
258
,
7260
7263
Colca
J. R.
,
Wolf
B. A.
,
Comens
P. G.
,
McDaniel
M. L.
(
1985
).
Protein phosphorylation in permeabilized pancreatic islet cells.
Biochem. J
228
,
529
536
Creutz
C. E.
,
Moss
S.
,
Edwadson
J. M.
,
Hide
I.
,
Gomperts
B.
(
1992
).
Differential recognition of secretory vesicles by annexins. (European Molecular Biology Organization Course: Advanced Techniques for Studying Secretion.).
Biochem. Biophys. Res. Commun
184
,
347
352
Drust
D. S.
,
Creutz
C. E.
(
1989
).
Aggregation of chromaffin granules by calpactin at micromolar levels of calcium.
Nature
331
,
88
91
Emans
N.
,
Gorvel
J. P.
,
Walter
C.
,
Gerke
V.
,
Kellner
R.
,
Griffiths
G.
,
Gruenberg
J.
(
1993
).
Annexin II is a major component of fusogenic endosomal vesicles.
J. Cell Biol
120
,
1357
1369
Fava
R. A.
,
Cohen
S.
(
1984
).
Isolation of a calcium-dependent 35-kilodalton substrate for the epidermal growth factor receptor/kinase from A-431 cells.
J. Biol. Chem
259
,
2636
2645
Futter
C. E.
,
Felder
S.
,
Schlessinger
J.
,
Ullrich
A.
,
Hopkins
C. R.
(
1993
).
Annexin I is phosphorylated in the multivesicular body during the processing of the epidermal growth factor receptor.
J. Cell Biol
120
,
77
83
Geisow
M. J.
,
Walker
J. H.
(
1986
).
New proteins involved in cell regulation by Ca2+and phospholipids.
Trends Biochem. Sci
11
,
420
423
Geisow
M. J.
,
Walker
J. H.
,
Boustead
C.
,
Taylor
W.
(
1987
).
Annexins- new family of Ca2+-regulated-phospholipid binding proteins.
Biosci. Rep
7
,
289
298
Glenny
J. R.
(
1986
).
Two related but distinct forms of the M r36000 tyrosine kinase substrate (calpactin) that interact with phospholipid and actin in a Ca2+-dependent manner.
Proc. Nat. Acad. Sci. USA
83
,
4258
4262
Haigler
H. T.
,
Schlaepfer
D. D.
,
Burgess
W. H.
(
1987
).
Characterization of lipocortin I and an immunologically unrelated 33-kDa protein as epidermal growth factor receptor/kinase substrates and phospholipase A2inhibitors.
J. Biol. Chem
262
,
6921
6930
Handel
S. E.
,
Rennison
M. E.
,
Wilde
C. J.
,
Buygoyne
R. D.
(
1991
).
Annexin II (calpactin I) in the mouse mammary gland:immunolocalization by light-and electron microscopy.
Cell Tiss. Res
264
,
549
554
Harrison
D. E.
,
Ashcroft
S. J. H.
(
1982
).
Effect of Ca2+, calmodulin, and cyclic AMP on phosphorylation of endogenous proteins by homogenates of rat islets of Langerhans.
Biochim. Biophys. Acta
714
,
313
319
Heinze
E.
,
Schatz
H.
,
Nierle
C.
,
Pfeiffer
E. F.
(
1975
).
Insulin biosynthesis in isolated pancreas islets of fetal and newborn rats.
Diabetes
24
,
373
377
Iida
H.
,
Nishitani
H.
,
Shibata
Y.
(
1992
).
Protein p67. A calcium-binding protein localized at the sarcolemma of secretory atrial myocytes.
Circ. Res
70
,
370
381
Khanna
N. C.
,
Tokuda
M.
,
Waisman
D. M.
(
1986
).
Phosphorylation of lipocortins in vitro by protein kinase C.
Biochem. Biophys. Res. Commun
141
,
547
554
Khanna
N. C.
,
Tokuda
M.
,
Waisman
D. M.
(
1987
).
Purification of three forms of lipocortin from bovine lung.
Cell Calcium
8
,
217
228
Klee
C. B.
(
1988
).
Ca2+-dependent phospholipid-(and membrane-) binding proteins.
Biochemistry
27
,
6645
6653
Korbutt
G. S.
,
Pipeleers
D. G.
(
1992
).
Cold storage of rat pancreas before purification of islet-cells.
Diabetes
41
,
299
307
Kuijpers
G. A.
,
Lee
G.
,
Pollard
H. B.
(
1992
).
Immunolocalization of synexin (annexin VII) in adrenal chromaffin granules and chromaffin cells: evidence for a dynamic role in the secretory process.
Cell Tiss. Res
269
,
323
330
Landt
M.
,
Eason
R. A.
,
Colca
J. R.
,
Wolf
B. A.
,
Turk
J.
,
Mills
L. A.
,
McDaniel
M. L.
(
1992
).
Parallel effects of arachidonic acid on insulin secretion, calmodulin-dependent protein kinase activity and protein kinase C activity in pancreatic islets.
Cell Calcium
13
,
163
172
Load
J. M.
,
Ashcroft
S. J. H.
(
1984
).
Identification and characterization of Ca2+-phospholipid-dependent protein kinase in rat islets and hamster-cells.
Biochem. J
219
,
547
551
McEvoy
R. C.
(
1981
).
Changes in the volumes of the A-, B-, and D cell populations in the pancreatic islets during the postnatal development of the rat.
Diabetes
30
,
813
817
Meers
P.
,
Mealy
T.
,
Tauber
A. I.
(
1993
).
Annexin I interactions with human neutrophil specific granules: fusogenicity and coaggregation with plasma membrane vesicles.
Biochim. Biophys. Acta
1147
,
177
184
Metz
S. A.
(
1988
).
Perspectives in diabetes. Is protein kinase C required for physiologic insulin release?.
Diabetes
37
,
3
7
Michener
M. L.
,
Dawson
W. B.
,
Creutz
C. E.
(
1986
).
Phosphorylation of a chromaffin granule-binding protein in stimulated chromaffin cells.
J. Biol. Chem
261
,
6548
6555
Nakata
T.
,
Sobue
K.
,
Hirokawa
N.
(
1990
).
Conformational change and localization of calpactin I complex involved in exocytosis as revealed by quick-freeze, deep-etch electron microscopy and immunocytochemistry.
J. Cell Biol
110
,
13
25
Overholser
M. D.
(
1925
).
The number of islets of Langerhans in the pancreas of the albino rat.
Endocrinology
9
,
493
504
Pepinsky
R. B.
,
Tizard
R.
,
Mattaliano
R. J.
,
Sinclair
L. K.
,
Miller
G. T.
,
Browning
J. L.
,
Chow
E. P.
,
Burne
C.
,
Huang
K.-S.
,
Pratt
D.
,
Wachter
L.
,
Hession
C.
,
Frey
A. Z.
,
Wallner
B. P.
(
1988
).
Five distinct calcium and phospholipid proteins share homology with lipocortin I.
J. Biol. Chem
263
,
10799
10811
Roth
D.
,
Morgan
A.
,
Burgoyne
R. D.
(
1993
).
Identification of a key domain in annexin and 14–3-3 proteins that stimulate calcium-dependent exocytosis in permeabilized adrenal chromaffin cells.
FEBS Lett
320
,
207
210
Sawyer
S. T.
,
Cohen
S.
(
1985
).
Epidermal growth factor stimulates the phosphorylation of the calcium-dependent 35, 000-Dalton substrate in intact A-431 cells.
J. Biol. Chem
260
,
8233
8236
Schlaepfer
D. D.
,
Haigler
H. T.
(
1988
).
In vitro protein kinase C phosphorylation sites of placental lipocortin.
Biochemistry
27
,
4253
4258
Schlaepfer
D. D.
,
Haigler
H. T.
(
1990
).
Expression of annexins as a function of cellular growth state.
J. Cell Biol
111
,
229
238
Schlaepfer
D. D.
,
Fisher
D. A.
,
Brandt
M. E.
,
Bode
H. R.
,
Jones
J. M.
(
1992
).
Identification of a novel annexin in hydra vulgaris.
J. Biol. Chem
267
,
9529
9539
Summers
T. A.
,
Creutz
C. E.
(
1985
).
Phosphorylation of a chromaffin granule-binding protein by protein kinase C.
J. Biol. Chem
260
,
2437
2443
Thams
P.
,
Capito
K.
,
Hedeskov
C. K.
(
1984
).
Endogenous substrate proteins for calcium-calmodulin-dependent, Ca2+-phospholipid-dependent and cyclic AMP-dependent protein kinases in mouse pancreatic islets.
Biochem. J
221
,
247
253
Turgeon
J. L.
,
Cooper
R. H.
,
Waring
D. W.
(
1990
).
Membrane-specific association of annexin I and annexin II in anterior pituitary cells.
Endocrinology
128
,
96
102
Wang
W.
,
Creutz
C. E.
(
1992
).
Regulation of chromaffin granule aggregating activity of annexin I by phosphorylation.
Biochemistry
31
,
9934
9939
Watkins
D. T.
(
1991
).
Ca2+-calmodulin-dependent phosphorylation of islet secretory granule proteins.
Diabetes
40
,
1063
1068
Zokas
L.
,
Glenny
J. R.
(
1987
).
The calpactin light chain is tightly linkedto the cytoskeletal form of calpactin I: studies using monoclonal antibodies to calpactin subunits.
J. Biol. Chem
105
,
2111
2121
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