219 DISC

Veterinary Dermatology 2001, 12, 8187

Cytokeratins in the canine epidermis

JAKOB H. WALTER
Institute of Veterinary Pathology, Freie Universitat Berlin, Strasse 518 Nr. 15, 14163 Berlin, Germany
(Received 6 April 1999; accepted 24 November 1999)

Abstract The purpose of this study was to characterize the cytokeratins (CKs) present in the clinically
normal skin of dogs. Skin samples from ve German shepherds, ve Boxers, ve Cocker spaniels, ve
Yorkshire terriers and ve mongrels were examined biochemically (using gel electrophoresis and western
blotting) and immunohistochemically (using a alkaline phosphatase anti-alkaline phosphatase technique).
Results indicated that the canine epidermis expressed the cytokeratins 1, 5, 6, 10/11, 14 and 16. There were no
consistent dierences in CK expression between the examined breeds with the exception of an individual
polymorphism in CK1 and CK10/11. Immunohistochemical studies showed CK 14 labelling of the basal cell
layer whereas CK10/11 staining was seen in the suprabasal cell layer of epidermis. Surprisingly, expression of
CK6, known as `stress' cytokeratin, was demonstrated in all epidermal samples. These results indicate that
there is a striking consistency of cytokeratin expression in dierent breeds which should be useful in the
investigation and characterization of canine skin diseases.
Keywords: cytokeratin, dogs, epidermis, keratinocytes, skin.

INTRODUCTION
Cytokeratins (CKs) are components of the intermediate lament network of epithelial cells. Up to
85% of the total content of the keratinocytes is
composed of cytokeratins.1 At the time of writing, 20
dierent cytokeratins (excluding hair keratin) have
been identied in humans,2 which shows a remarkable epithelial tissue specic expression.3 For the
human epidermis it has been shown that the
traditional Moll catalogue3,4 must be supplemented
with polymorphic and individual CK variations.58
Although CKs have been described previously in
many immunohistochemical studies of canine skin,
especially in studies of neoplasia,912 fundamental
experiments on the composition of cytokeratins in
normal dog skin have not yet been carried out. The
aim of this study was to characterize the cytokeratins
and possible polymorphisms in the normal canine
epidermis which may be of value in the dierential
diagnosis of inammatory, inherited, pre-malignant
and malignant diseases of skin.
MATERIALS AND METHODS
Unless otherwise stated, all chemicals were obtained
from Sigma, Deisenhofen, Germany.
Samples
Skin samples (each approximately 262 cm) were

Correspondence: JH Walter, VetPathol., Strasse 518 Nr. 15, 14163

Berlin, Germany. E-mail: jhwpatho@zedat.fu-berlin.de
# 2001 Blackwell Science Ltd

taken from the lateral thorax of dogs with clinically

normal skin from a selection of dierent adult breeds
(ve German shepherds, ve Boxers, ve Cocker
spaniels, ve Yorkshire terriers, ve mongrels) which
were presented to a veterinary hospital (Klinik u.
Poliklinik f. Kleine Haustiere, FU Berlin). Nine of
the dogs were killed in trac accidents and the
samples were taken 418 h after death. Sixteen of the
dogs were euthanased because of various disorders
(nine cases of canine congestive cardiomyopathy,
four cases of behavioural disorders, three cases of
diseases of the lower respiratory system) and the
samples were taken directly post mortem.
The skin samples were divided: one half was used
immediately for the preparation of the epidermal
cytokeratins. The other half was xed in 10% neutral
formalin for 24 h, embedded in paran wax (DDMTissue Embedding Medium; Medim, Giessen, Germany) and sectioned (4 mm). The sections were stained
routinely with haematoxylin and eosin (HE) or
processed for immunohistochemistry using the Sequenza system, a disposable coverplate incubation system
(Shandon; Frankfurt/Main, Germany). Histopathological examination of HE-stained skin sections used in
this study showed no histopathological abnormalities.
Cytokeratin preparation and separation
The skin sample was placed in a water bath at 608C
for 5 min and the epidermis was then peeled o the
dermis using the back of a scalpel blade. Cytokeratins
were extracted from the epidermis in accordance with
the methods described previously.13 The epidermal
material (0.5 g) peeled from the skin was minced into
smaller pieces and homogenized in 2 mL of homogenizing buer containing 96 mM NaCl, 8 mM
81

Ahed
Bhed
Ched
Dhed
Ref marker
Fig marker
Table
marker
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219 DISC
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J.H. Walter

KH2PO4, 5.6 mM Na2HPO22H2O, 1.5 mM KCl, 10 mM

ethylenediaminetetraacetate (EDTA) and 0.1 mM dithiothreitol (DTT) in a homogenisator (Braun-Potter;
Melsungen, Germany) at 400 g for 3 min. The homogenized material was divided into 100 mL aliquots,
which were placed in an extraction buer (10 mM TrisHCl, 140 mM NaCl, 5 mM EDTA, 1.5 M KCl, 5 mM
DTT and 1% Triton X-100) at 48C for 30 min. After
centrifugation (10 000 g at 48C for 3615 min, the
supernatant was discarded and the pellet was boiled in
sample buer (2.5% SDS, 10 mM Tris-HCl, 10%
glycerol, 50 mM DTT) for 15 min. The extracted material was subjected to one-dimensional gel electrophoresis in dilutions of 1: 1, 1: 5, 1: 10, and 1: 15 in sample
buer. Sodium dodecyl sulphate-polyacrylamide gel
electrophoresis (SDS-PAGE) was performed on two
12.5% gels with acrylamide and bisacrylamide in a
ratio of 200: 1. 14 Samples of 18 mL each were run in
four lanes of each gel together with a molecular weight
marker mixture (Dalton Marker VII-L; Sigma). One
gel was stained with Coomassie Blue for 1 h, and destained for 1218 h. Then it was laid between 2 sheets of
transparent lm and digitalized with a scanner (Scanjet
II cx, Hewlett Packard, Boeblingen, Germany).

nonspecic binding. The membrane strips were

washed three times in TBS with 1% Tween 20 and
incubated with the primary antibodies at appropriate
dilutions (Table 1) into 3% BSA in TBS also
containing 0.5% Tween 20 for 18 h at 48C. The
strips were washed three times in TBS and an
alkaline-phosphatase conjugated goat antimouse
antibody (Medac; Hamburg, Germany) was added
to each strip in a dilution of 1 : 1000 in TBS
containing 0.5% Tween 20 at 378C for 120 min.
The membrane strips were washed as before and the
reaction was visualized using a solution of 0.6 mg
mL71 Fast Red Salt and 4 mg mL71 naphthol ASMX phosphate in TBS by incubation at 378C for 30
min. After a nal washing in distilled water, the strips
were dried and were digitalized with an HP Scanjet II
cx. The digitalized results were evaluated on a
Macintosh computer (Quadra 660 AV) with the help
of the public domain program NIH Image (developed
at the U.S. National Institute of Health, available on
the Internet at http://rsb.info.nih.gov/nih-image/).
Immunohistochemistry
A slightly modied alkaline phosphatase anti-alkaline phosphatase (APAAP)-technique based on that
described by Cordell et al.16 was used for the
immunohistochemical studies. The APAAP reagent
was taken from a commercially available APAAP-kit
(Dianova, Hamburg, Germany). Four mm-thick
deparanized sections were heated in a microwave
oven (265 min at 600 W) in 10 mM citrate buer, pH
6.0. Sections were rinsed three times in TBS pH 7.4,
containing 0.05 M Tris-HCl and 0.5 M NaCl and the
following tests were carried out in the Sequenza
staining station (Shandon). The sections were incubated with primary antibody (Table 1) diluted in
Roswell Park Memorial Institute cell culture medium
(RPMI) adjusted to pH 7.4 for 30 min at room
temperature. After being washed with TBS, the
secondary antibody (diluted 1 : 100 in RPMI pH 7.4
containing 5% normal dog serum) was added for 30
min. After one further wash, sections were incubated
with the substrate (prepared by dissolving 20 mg
naphtol-AS-MX-phosphate in 2 mL N,N-dimethyl-

Antibodies
Thirteen monoclonal antibodies (MAbs) raised
against various human cytokeratins were used, which
are listed in Table 1.
Immunoblotting
The second gel was equilibrated in transfer buer
(0.04 M 6-aminohexane acid, 0.001 M sodium dodecyl
sulphate, 20% methanol) and the separated polypeptides were transferred electrically to nitrocellulose
membrane (Millipore Filters, Type HA, pore size 0.45
mm) in a blotting chamber.15 As a control for the
protein transfer, the membranes were stained with
0.5% Ponceau-S. The visualisized lanes were cut out
in strips, and the colour washed out in distilled water.
The membrane strips were soaked in 3% bovine
serum albumin (BSA) in Tris Based Saline (TBS)
containing 0.01 M Tris-HCl pH 7.6 and 0.5 M NaCl
for 2630 min at room temperature to block

Table 1. Monoclonal antibodies for immunohistochemistry and western blot

Antibody

Cytokeratin specity

Dilution APAAP

Dilution western blot

Source

AE1
AE3
KL1
CAM 5.2
LP 34
Ks 7.18
DE-K10
AE8
LL002
E3
Ks 18.04
Ks 19.1
CK 20

10,14, 16, 19
1, 5, 8
10/11
8, 18
6, 18
7
10
13
14
17
18
19
20

1: 400
1: 400
1: 100
ready to use
1: 100
1: 50
1: 10
1: 20
1: 20
1: 20
1: 10
1: 10
1: 10

1:
1:
1:
1:
1:
1:
1:
1:
1:
1:
1:
1:
1:

ICN; Meckenheim, Germany

ICN; Meckenheim, Germany
Immunotech; Hamburg, Germany
Becton Dickinson; Heidelberg, Germany
Dako; Hamburg, Germany
Progen; Heidelberg, Germany
Dako; Hamburg, Germany
BioGenex; Hamburg, Germany
Serotec; Wiesbaden, Germany
Dako; Hamburg, Germany
Progen; Heidelberg, Germany
Progen; Heidelberg, Germany
R. Moll; Mainz, Germany

# 2001 Blackwell Science Ltd, Veterinary Dermatology, 12, 8187

1000
1000
1500
10
1500
1000
1500
200
200
400
400
200
100

219 DISC
Cytokeratins in the canine epidermis

83

formamide, adding 98 mL Tris buer (0.05 M, pH

8.2), 100 mg Fast Red TR and 100 mL 1 M levamisole)
for 30 min. As positive controls in the preliminary
part of the experiment, similarly treated sections of
human skin were used. For a negative control,
consecutive sections of canine skin were included in
each experiment which had the primary antibody
replaced by a nonimmunogenic mouse serum (Dianova). The immune reaction was evaluated semiquantitatively. The relative proportion of immunoreactive cells was determined in 10 microscopic elds
at 2506 magnication and `scored' as follows: []: no
immune reactive cells; [1 + ]: 10% immune reactive
cells; [2 + ]: 1150% immune reactive cells; [3 + ]:
5175% immune reactive cells; [4 + ]: _ 75% immune
reactive cells. A score of 1 + was regarded as
insucient evidence of immunostaining.

kDa) were visible in all SDS-PAGE gels (Fig. 1). In

the case of one Boxer, an additional 37.5 kDa band of
uncertain origin appeared.

RESULTS

Immunohistochemistry
Also when using immunohistochemical staining, only
ve of the antibodies used showed consistently
positive immunostainings, while the other antibodies
gave negative results:
. AE1 with a [4 + ] reaction (Fig. 4)
. KL1 with a [4 + ] reaction
. LL002 with a [4 + ] reaction (Fig. 5)
. LP34 with a [4 + ] reaction (Fig. 6)
. AE3 with a [3 + ] reaction.
These antibodies reacted in the epidermis with the
following results (see also Table 3): LL002 (CK14;
Fig. 5) stained only the basal cell layer, basal and
suprabasal cells were marked by LP34 (CK6/18; Fig.
6) and AE3, while KL1 (CK10/11) and AE1 (Fig. 5)
were only immunoreactive in the suprabasal cells.
Immunostaining of human skin sections (positive
controls) gave appropriate results throughout, while
the negative controls were consistently negative.

SDS-PAGE
In all the 25 skin samples examined, ve bands could
be identied clearly by SDS-PAGE. The molecular
weight (MW) of these bands was 68, 58, 56, 53 and 50
kiloDalton (kDa) (Fig. 1) and these molecular
weights correspond with the cytokeratins CK1,
CK5, CK6, CK10/11, CK14 and CK16 in the human
cytokeratin catalogue.3 Minimal molecular weight
dierences in CK1 and CK5 appear to indicate
individual variations. In some individual cases, a
double peak was identied in CK1 and/or CK10/11
(Fig. 2). A double peak was found in CK1 and/or
CK10 in two German shepherds, four Boxers, one
Cocker spaniel, four Yorkshire terriers and four
mongrels (Table 2). There were no consistent
dierences between the breeds. In addition a prominent band of 32 kDa, and three bands (MW 4 21

Western blot
The immunoblots showed that the following antibodies recognize distinct cytokeratins in the canine
epidermis: AE1 identies CK10, 14 and 16, AE3
identies CK1 and CK5, KL1 identies CK 10/11,
LP34 identies CK6 and LL002 identies CK14. The
prominent 30 kDa band, the three bands with MW's
4 21 kDa and the additional 37.5 kDa band were not
immunoreactive with the antibodies used.
As on SDS-PAGE, the CK6 and CK10/11 bands
both shared the same mobility, the individual
presence of CK6 and/or CK10 could only be proved
by western blot, after staining with LP34 or KL1
antibodies (Fig. 3).

Figure 1. Representative SDSPAGE of normal canine

epidermis: (a) Coomasie Blue
stained gel showing separation
of four serial dilutions of the
same protein sample and of
marker proteins (M); (b)
Densitoblot of marker proteins
(M) generated by NIH Image;
(c) Densitoblot (NIH Image) of
the epidermal proteins; the
calculated molecular weights (in
kDa) correspond to cytokeratins
1, 5, 6 (10/11 resp.), 14 and 16
(from top). There are bands at
30, 21, 15.5 and 14.5 kDa of
unknown origin (see Discussion).
# 2001 Blackwell Science Ltd, Veterinary Dermatology, 12, 8187

219 DISC
84

J.H. Walter
Figure 2. Polymorphism of the cytokeratins of the
epidermis: (a) Variation in CK1 with double peak
(*CK1a/b); (b) Variation in CK10 with double peak
(**CK10a/b); and (c) Variation in CK1 and CK10
(CK1a/b and CK10a/b) 12.5% SDS-PAGE;
Coomassie Blue.

Table 2. Incidence of polymorphic cytokeratins on SDS-PAGE of extracts of epidermis from dierent breeds
4 Breed

CK1a/1b

CK 10a/10b

CK 1a/1b & 10a/10b

No polymorphism

4 German Shepherd
4 Boxer
Cocker Spaniel
Yorkshire Terrier
4 Mixed breed
Total

5
5
5
5
5
25

0
3
1
3
3
10

0
1
0
1
1
3

2
0
1
0
1
4

3
1
3
1
0
8

Figure 3. Immunoblots demonstrating the presence

of both cytokeratins 6 and 10: (a) SDS-PAGE of the
epidermis; Western blot labelled with (b) KL1
(CK10/11) and (c) LP34 (CK6) MAbs.

Figure 4. Immunohistochemical staining of the basal

cells of the canine epidermis with LL002 (CK14)
MAb, APAAP-Fast Red.

Figure 5. Immunohistochemical staining by AE 1

Mab of suprabasal cells of the canine epidermis,
APAAP-Fast Red.

DISCUSSION
The composition of the cytokeratins in the epidermis
of dogs appears to have a similar polymorphism to
# 2001 Blackwell Science Ltd, Veterinary Dermatology, 12, 8187

that of humans.5,6,8,17 In spite of the many breeds that

have exhibited morphological dierences in the
epidermis,18 the results of SDS-PAGE show polymorphic variations between individuals (at least for

219 DISC
Cytokeratins in the canine epidermis

85

Figure 6. Immunohistochemical staining of canine

epidermis by LP 34 MAb. All the cell layers of the
epidermis demonstrate intensive staining, APAAPFast Red.

Table 3. Immunohistochemical reactivity of the canine epidermis

Antibody

basal layer

suprabasal layer

AE1
AE3
KL1
CAM 5.2
LP 34
Ks 7.18
DE-K10
AE8
LL002
E3
Ks 18.04
Ks 19.1
CK 20

+
+
+

+ indicates consistent staining

the cytokeratins 1 and 10) and were observed in all the

breeds examined. The evidence of CK6 in normal
epidermis from all the dogs examined in this study is
surprising, as this was not observed in human skin
specimens. In the human epidermis, the CK-expression is characterized by a clear division between the
basal and suprabasal layers. Cytokeratins 5, 14 and 15
are found in the basal layer and the CK1 and 10 in the
suprabasal layer of the human epidermis.2 CK6 and
16 only appear in hyperproliferative skin processes or
in areas of modied epidermis such as the sole of the
foot.1921 This probably is connected to the reduced
expression of CK1 and CK10.20 Cytokeratins in
especially stressed epidermis need, it seems, an
additional stabilization factor and that is probably
the function of CK6 and CK16.22 In contrast, the dog
epidermis consists of 36 layers and is 1/3 as thick as
the human epidermis.18 It has been shown that canine
skin receives as much load or stress23,24 as human
skin, and it is possible that the dierent cytokeratin
expression reects adaptation to resist these loads.
The results of this study show that cytokeratins 1, 5, 6
and 14 are expressed by basal cells and cytokeratins 4,
10/11, 14 and probably 15/16 by the suprabasal cells. As
on SDS-PAGE, cytokeratin 6 and 10 have the same
mobility, the presence of CK6 was conrmed by
western blot immunoreactivity with LP34. The cytokeratin composition of the epidermis of the dog appears
to be more complex than that of humans and, although

it is thinner, it probably has equal strength.23,24 On the

basis of immunohistochemical staining patterns, LP34
MAb or the combination of AE 1/AE 3 MAbs can be
recommended as pan-epidermal markers.
The additional `cytokeratin' band observed in one
Boxer could represent a degradation product of
cytokeratin, the epitope of which was not recognized
by the antibodies used.5 In this case, possible postmortem decomposition can be excluded because this
skin sample was taken directly after euthanasia.
According to their molecular weights (MW), the
unknown prominent band with 30 kDa MW probably
resembles llagrin 25 and the three additional bands
with 21, 15.5 and 14.5 kDa MW resemble histones.26
This study is an additional step in the development of a
catalogue of canine cytokeratins. Because gene sequences
for canine intermediate laments are not yet known
(based on a computer search at the GenBank of the
National Center for Biotechnology Information [http://
www.ncbi.nlm.nih.gov/Genbank]), further experiments
and methods are necessary to compile a nal inventory.
The ndings of this study may have clinical and
histopathological value in dierent skin diseases. For
example, they may be relevant in the denitive diagnosis
of undierentiated epidermal neoplasias, because the
cytokeratin expression exhibits a tissue specicity that is
retained in neoplastic transformation.2,3 Further studies
of the pattern of the expression of cytokeratins in nonneoplastic skin lesions are also be recommended,
because dierent studies in mouse models suggest that
chronic inammation may induce changes in cytokeratin expression27 and that abnormalities associated with
`keratinopathies' result from cytokeratin changes.28
ACKNOWLEDGEMENTS
I am grateful to Susanne Hahn for expert technical
assistance.
REFERENCES
1. Fuchs, E. Keratins and the skin. Annual Review of Cell
and Developmental Biology 1995; 11: 12353.
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J.H. Walter

2. Moll, R., ed. Cytokeratine als Dierenzierungsmarker:

Expressionsprole von Epithelien und epithelialen
Tumoren. Stuttgart: Gustav Fischer Verlag, 1993.
3. Moll, R., Franke, W.W., Schiller, D.L., Geiger, B.,
Krepler, R. The catalog of human cytokeratins:
patterns of expression in normal epithelia, tumors
and cultured cells. Cell 1982; 31: 1124.
4. Moll, R., Schiller, D.L., Franke, W.W. Identication of
protein IT of the intestinal cytoskeleton as a novel type
I cytokeratin with unusual properties and expression
patterns. Journal of Cell Biology 1990; 111: 56780.
5. Celis, J.E., Rasmussen, H.H., Olsen, E., Madsen, P.,
Leers, H., Honore, B. et al. The human keratinocyte
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proliferation, dierentiation and skin diseases.
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6. Korge, B.P., Gan, S.-Q., McBride, O.W., Mischke, D.,
Steinert, P.M. Extensive size polymorphism of the
human keratin 10 chain resides in the C-terminal V2
subdomain due to variable numbers and sizes of glycine
loops. Proceedings of the National Academy of Sciences
of the United States of America 1992; 89: 9104.
7. Lobeck, H., Bartke, I., Naujoks, K., Muller, D.,
Bronhoft, G., Mischke, D., et al. Verteilungsmuster
der Zytokeratinpolypeptide 4 und 5 Im Normalen und
Neoplastischen Epithel Unter Verwendung Neuer
Parangangiger Monoklonaler Antikorper (Eine
Immunhistochemische Untersuchung). Verhandlungen
der Deutschen Gesellschaft fur Pathologie 1989; 73: 645.
8. Mischke, D., Wild, G. Polymorphic keratins in human
epidermis. Journal of Investigative Dermatology 1987;
88: 1917.
9. Andreasen, C.B., Mahaey, E.A., Duncan, J.R.
Intermediate lament staining in the cytologic and
histologic diagnosis of canine skin and soft tissue
tumors. Veterinary Pathology 1988; 25(5): 3439.
10. Cardona, A., Madewell, B.R., Naydan, D.K., Lund,
J.K. A comparison of six monoclonal antibodies for
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canine tissues. Journal of Veterinary Diagnostic
Investigation 1989; 1: 31623.
11. Ferrer, L., Rabanal, R.M., Fondevila, D., Prats, N.
Immunocytochemical demonstration of intermediate
lament proteins, S-100 protein and CEA in apocrine
sweat glands and apocrine gland derived lesions of the
dog. Journal of Veterinary Medicine Series A 1990;
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12. Rabanal, R.H., Fondevila, D., Montane, V., Domingo,
M., Ferrer, L. Immunocytochemical diagnosis of skin
tumours of the dog with special reference to
undierentiated types. Research in Veterinary Science
1989; 47: 12933.
13. Achtstaetter, T., Hatzfeld, M., Quinlan, R.A., Parmelee,
D.C., Franke, W.W. Separation of cytokeratin
polypeptides by gel electrophoretic and chromatographic
techniques and their identication by immunoblotting.
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14. Laemmli, U.K. Cleavage of structural proteins during
the assembly of the head of bacteriophage T4. Nature
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15. Towbin, H., Staehlin, T., Gordon, J. Electrophoretic

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16. Cordell, J.L., Falini, B., Erber, W.N., Ghosh, A.K.,
Abdulaziz, Z., MacDonald, S. et al. Immunoenzymatic
labelling of monoclonal antibodies using immune
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18. Schwarz, R., Le Roux, J.M.W., Schaller, R., Neurand,
K. Micromorphology of the skin (epidermis, dermis,
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Veterinary Research 1979; 46: 1059.
19. Moll, I., Moll, R. Cells of extramammary Paget's
disease express cytokeratins dierent from those of
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20. Stoler, A., Kopan, R., Duvic, M., Fuchs, E. Use of
monospecic antisera and cRNA probes to localize the
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21. Weiss, R.A., Eichner, R., Sun, T.T. Monoclonal
antibody analysis of keratin expression in epidermal
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marker for hyperproliferative keratinocytes. Journal of
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22. Jiang, C.-K., Magnaldo, T., Ohtsuki, M., Freedberg,
I.M., Bernerd, F., Blumenberg, M. Epidermal growth
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Pathobiology 1999; 67: 4550.
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Cytokeratins in the canine epidermis

87

Resume Le but de cette etude etait de determiner le type de cytokeratines presentes dans la peau du chien.
Des prelevements cutanes, realises sur cinq Bergers allemand, cinq Boxers, cinq Cocker Spaniel, cinq
Yorkshire terrier et cinq croises ont ete examines par biochimie (electrophorese en gel et western blotting) et
par immunohistochimie (technique alkaline phosphatase anti-alkaline phosphatase). Les resultats ont montre
que l'epiderme canin exprime les cytokeratines 1, 5, 6, 10/11, 14 et 16. Aucune dierence n'a ete observee entre
les races examinees, a l'exception d'un polymorphisme individuel pour l'expression de CK1 et de CK10/11.
Les etudes immunohistochimiques ont montre un marquage de la couche basale par CK 14 alors qu'un
marquage par CK10/11 etait observe dans les couches suprabasales. Il a ete, etonnamment, observe une
expression constante de CK6, connue comme une cytokeratine de `stress' dans tous les prelevements
epidermiques. Ces resultats indiquent qu'il existe une similarite d'expression des cytokeratines dans dierentes
races canines, ce qui devrait etre utile pour la recherche et la caracterisation des dermatoses chez le chien.
[Walter, J. H. Cytokeratins in the canine epidermis. (Cytokeratines de l'epiderme du chien.) Veterinary
Dermatology 2001; 12: 8187.]
Resumen El objetivo de este estudio fue caracterizar las citoqueratinas (CKs) presentes en piel cl nicamente
normal del perro. Se examinaron muestras cutaneas de cinco perros Pastor Aleman, cinco Boxers, cinco
Cocker spaniels, cinco terriers de Yorkshire y cinco perros mestizos mediante pruebas bioqu micas (usando
electroforesis con gel y western blotting) e inmunohistoqu micamente (usando una tecnica de fosfatasa
alcalina anti-fosfatasa alcalina). Los resultados indicaron que la epidermis canina expresaba las citoqueratinas
1, 5, 6, 10/11, 14 y 16. No exist an diferencias signicativas en la expresion de CK entre las razas examinadas
con la excepcion de un polimorsmo individual en la CK1 y CK10/11. Los estudios inmunohistoqu micos
mostraron marcaje con CK 14 en la capa de celulas basales mientras que se observo tincion con CK10/11 en la
capa de celulas suprabasales de la epidermis. Sorprendentemente, la expresion de CK6, conocida como
citoqueratina de `stress', se observo en todas las muestras de epidermis. Estos resultados indican que existe
una destacada constancia en la expresion de citoqueratina en diferentes razas, que deber a ser de utilidad en la
investigacion y caracterizacion de enfermedades cutaneas caninas. [Walter, J. H. Cytokeratins in the canine
epidermis. (Citoqueratina en la epidermis canina.) Veterinary Dermatology 2001; 12: 8187.]
Zusammenfassung Der Zweck dieser Studie war die Charakerisierung der in der klinisch normalen Haut der
Hunde vorhandenen Zytokeratine (ZK). Hautproben von funf deutschen Schaferhunden, funf Boxern, funf
Cockerspanieln, funf Yorkshireterriern und funf Mischlingen wurden biochemisch (mittels Gelelektrophorese
und Western Blot) und immunhistochemisch (mittels einer alkalischen anti-alkalischen Phosphatase Technik)
uberpruft. Resultate zeigten, dass Hundeepidermis die Zytokeratine 1, 5, 6, 10/11, 14 und 16 exprimierte. Mit
Ausnahme einer individuellen Polymorphie von ZK1 und ZK10/11 gab es keine Unterschiede der ZKExprimierung zwischen den untersuchten Rassen. Immunhistochemische Studien zeigten ZK 14 Markierung
in der Basalzellschicht, wahrend ZK10/11 Farbung in den suprabasalen Zellen der Epidermis gesehen wurde.
Uberraschend wurde das als Stresszytokeratin bekannte Zk6 in allen epidermalen Proben nachgewiesen. Diese
Resultate zeigen eine auallende Ubereinstimmung der Zytokeratinexprimierung bei verschiedenen Rassen,
die bei der Untersuchung und der Charakterisierung von Hautkrankheiten beim Hund nutzlich sein sollte.
[Walter, J. H. Cytokeratins in the canine epidermis. (Zytokeratine in der Epidermis des Hundes.) Veterinary
Dermatology 2001; 12: 8187.]

# 2001 Blackwell Science Ltd, Veterinary Dermatology, 12, 8187