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Oestrogen receptor evaluation in Pomeranian dogs with

hair cycle arrest (alopecia X) on melatonin supplementation


Blackwell Publishing Ltd

Linda A. Frank*, Robert L. Donnell and


Stephen A. Kania
*Department of Small Animal Clinical Sciences
Department of Pathobiology
Department of Comparative Medicine, University of Tennessee,
Knoxville, Tennessee, USA
Correspondence: Linda A. Frank, C247 Veterinary Teaching Hospital,
University of Tennessee, Knoxville, TN 37996-4544, USA
E-mail: lfrank@utk.edu
This project was funded by the American Kennel Club Canine Health
Foundation. The contents of this publication are solely the responsibility of
the authors and do not necessarily represent the views of the Foundation.

Abstract
The role of oestrogen receptors in dogs with hair cycle
arrest (alopecia X) was investigated by immunohistochemistry. The purpose of this study was to determine
if hair regrowth in dogs with hair cycle arrest treated
with melatonin was associated with a decrease in
follicular oestrogen receptors. Fifteen Pomeranians
(excluding intact females) with hair cycle arrest were
enrolled. Two biopsies were obtained from alopecic
areas of the trunk before and after 3 months on melatonin.
Haematoxylin and eosin-stained tissues were examined
was demonstrated immunoand oestrogen receptor-
histochemically. Common histopathological findings
included hyperkeratosis, follicular keratosis, excessive
tricholemmal keratinization (flame follicles), thin epidermis, few small anagen bulbs, epidermal pigmentation
and melanin aggregates within follicular keratin. Melanin
aggregates within basal cells and hair were an occasional
finding. After 3 months, 40% (six) dogs had mild to
moderate hair regrowth. Biopsies from six dogs showed
histological evidence of an increase in anagen hairs and
eight dogs had a decrease in epidermal pigmentation.
Moderate to marked staining intensity of oestrogen
was noted in all sebaceous gland basal cells,
receptor-
all small hair bulbs and follicular epithelium of telogen
staining of
hairs. There was no oestrogen receptor-
nuclei within the epidermis, apocrine glands or dermal
fibroblasts. Large anagen hair bulbs had minimal to no
oestrogen receptor staining. Hair regrowth was not asso staining.
ciated with a change in oestrogen receptor-
Accepted 21 March 2006

Introduction
Hair cycle arrest (also known as alopecia X, adrenal
hyperplasia-like syndrome, growth hormone responsive
252

alopecia, pseudo-Cushings syndrome, and others) is a


common condition seen in Pomeranians and other breeds
of dogs with dense undercoats that results in truncal
alopecia and hyperpigmentation of the skin.1,2 Systemic
illness is not associated with this disease.
The pathogenic mechanism of the alopecia is not
known. Diagnosis is based on clinical signs and ruling out
the inflammatory causes of alopecia (demodicosis, bacterial folliculitis and dermatophytosis) and the common noninflammatory causes of alopecia such as hypothyroidism
and hypercortisolemia (e.g. Cushings syndrome).1,2 Once
theorized to be associated with abnormalities of adrenal
steroid intermediates and sex hormones,3 it has been
recently shown that not all dogs with hair cycle arrest have
abnormalities in adrenal steroid or sex hormone concentrations and that there is no association of hair regrowth
with changes in hormone concentrations.4,5 It was originally hypothesized that melatonin exerted its effect on hair
growth through modulating hormone production;6 however, decreases in hormone concentrations were not
associated with hair regrowth in neutered dogs.5 Thus,
hair regrowth must be associated with another mechanism.
Little is known about the hair cycle and what controls it.
Of recent interest is the recognition of an oestrogen
receptor pathway that regulates the telogenanagen hair
follicle transition in mice.710 Oestrogen has been shown to
be produced by the hair follicle itself11 and oestrogen receptors
have been identified in canine hair follicles.12 Interestingly,
melatonin has been shown to block the activation of
oestrogen receptors in human breast cancer cells.13,14
We hypothesize that dogs with hair cycle arrest have
increased activity of oestrogen receptors in hair follicles,
preventing the hair follicles from cycling normally. Thus,
melatonin may cause hair regrowth in dogs with hair cycle
arrest by down-regulating the expression of oestrogen
receptors. The purpose of this study was to determine if
hair regrowth in dogs with hair cycle arrest when treated
with melatonin was associated with a decrease in follicular
oestrogen receptor staining.

Materials and methods


The study was approved by the University of Tennessee, Knoxville
Institutional Animal Care and Use Committee. Fifteen Pomeranian
dogs were enrolled in the study. Intact females were excluded from
enrolment. Three spayed female, seven castrated male and five intact
male Pomeranian dogs were entered between August 2004 and February 2005. The median age at time of enrolment was 5 years (range
4 6.6 years). Dogs were neutered a median of 2.25 years (range 1.5
5.5 years) prior to enrolment into the study. The median age of hair
loss was 1 year of age with a range of 0.5 5 years. Three of the neutered dogs were neutered before onset of hair loss, whereas seven
were neutered at some point following onset of hair loss. Eight dogs
had an orange coat colour, three had a black coat colour, two had a

2006 The Authors. Journal compilation 2006 European Society of Veterinary Dermatology. 17; 252258

Oestrogen receptors in dogs

cream coat colour and two had a blue coat colour. Dogs weighed
between 1.8 and 3 kg. The median melatonin dosage was 1.3 mg kg1
(range 11.7 mg kg1) every 12 h.
Dogs were evaluated by their regular veterinarian (n = 10) or at the
University Veterinary Teaching Hospital (n = 5). Prior to enrolment,
dogs were diagnosed with hair cycle arrest based on clinical signs
(alopecia was bilaterally symmetrical, sparing the head and forelimbs
and was not colour restricted or cyclical in nature), and hypothyroidism and hyperadrenocorticism have been ruled out. Hypothyroidism was excluded based on normal concentrations of thyroxine
and thyrotropin and/or lack of hair regrowth on appropriate thyroid
supplementation. Hypercortisolemia was excluded based on lack of
other clinicopathological abnormalities and cortisol concentrations
within the normal range following either low-dose dexamethasone
screening or adrenocorticotropic hormone stimulation.
Two biopsies were taken from alopecic areas of the trunk using a
6-mm biopsy punch and placed in 10% formalin. The biopsy samples
were mailed overnight, and were transferred into 70% EtOH within
24 h. Biopsies were embedded in paraffin and processed for routine
evaluation using haematoxylin and eosin (H&E). The tissues were
positioned in the paraffin blocks so that both longitudinal and horizontal sections of follicles could be obtained. The sections were microscopically evaluated to confirm the diagnosis of hair cycle arrest
(noninflammatory alopecia), and eliminate diseases that could mimic
this condition (sebaceous adenitis, colour dilution alopecia). A second
set of two biopsies were obtained from similar locations (but not previous biopsy sites) after 3 months on oral melatonin (Natures Bounty,
Bohemia, NY, USA) dosed at 3 mg dog1 every 12 h. This product has
been shown to contain at least 3 mg melatonin per tablet with good
oral bioavailability in dogs.15
Owners and veterinarians evaluated the dogs after 3 months of
oral melatonin for degree of hair regrowth and quality of new hair, if
any. A questionnaire was provided in which clinical description, percentage of body affected (< 25%, 2550%, 5075%, or > 75%) and
quality of hair (totally bald in affected areas, fine undercoat only in
affected areas, moderate undercoat with decreased guard hairs in
affected areas) were recorded. Mild hair regrowth was defined as no
change or only minimal change in percentage of body affected and
quality of new hair was only fine undercoat or sparse guard hairs.
Moderate hair regrowth was defined as a decrease in percentage of
body affected with moderate undercoat and guard hairs. No dog was
categorized as having complete hair regrowth because of the short
duration of the study. At that time a second set of biopsies samples
were obtained from each patient. If there was evidence of hair
regrowth, the biopsies were obtained from that region. The exact
locations of the biopsies were not recorded; however, all biopsies
were obtained from the trunk and biopsies were not obtained from
previous biopsy sites. Handling of the tissue was as described for the
initial biopsies.
Each set of biopsies were described together. The H&E-stained tissues were evaluated for presence of anagen and telogen hair follicles,
thickness of follicular and surface epithelium, presence of epidermal
and/or follicular hyperkeratosis, malformation of the follicles and presence of sebaceous and apocrine sweat glands.
Oestrogen receptors were detected immunohistochemically by
standard methods.16 Briefly, after dewaxing the slides were subjected
to antigen retrieval by steaming at 95 C for 25 min in a pH 6.1
retrieval solution (Dakocytomation Inc., Carpenteria, CA, USA), followed by rinsing in TBS. The slides were then processed on a DAKO
autostainer (Dakocytomation, Inc.) with 60-min incubation with the
mouse monoclonal NCL-ER-H2 antibody (Novocastra Laboratories
Ltd. Newcastle upon Tyne, UK) and completion of the process
employing HRP-labelled DakoEnvision+System reagents (Dakocytomation, Inc.) with the DAB + chromagen (Dakocytomation, Inc.) for
visualization of antibody-antigen binding.
Positively stained nuclei were counted (none = 0; < 25% = 1; 25
50% = 2; > 50% = 3) and the intensity of the reaction was assessed
(+ = mild; ++ = moderate; +++ = marked) within the nuclei of the follicular epithelium, hair bulb, epidermis, dermal fibroblasts and sweat
glands. Uterine tissue obtained from a female dog that was in heat
was used as the positive control. Samples from subjects with hair

regrowth, either from owner/veterinarian observation or from histopathological evidence (many anagen follicles) were compared to
those in which no hair growth was noted and to the initial biopsies for
each animal.

Results
Six (40%) of the dogs had mild to moderate hair regrowth
following 3 months of oral melatonin. Moderate hair
regrowth (defined as a decrease in percentage of body
affected with moderate undercoat and guard hairs) was
reported in three dogs (one orange castrated male, one
orange intact male, and one black spayed female). Mild
hair regrowth (defined as no change or only minimal
change in percentage of body affected and quality of new
hair was only fine undercoat or sparse guard hairs) was
also reported in three dogs (one orange intact male, one
black intact male, and one cream castrated male). No
dog was categorized as having complete hair regrowth
because of the short duration of the study. Owners of
three dogs with no hair regrowth reported a lightening of
the skin while on melatonin.
Histopathological evaluation
All initial biopsies were classified as having a noninflammatory alopecia. The histopathological findings are summarized in Table 1. All tissues showed hyperkeratosis and
follicular keratosis. Telogen hairs with prominent trichilemmal keratinization (flame follicles) were evident in all but
two dogs. The epidermis was 12 cell layers thick in all,
but one dog in which the epidermis was 34 cell layers
thick, and occasional follicles appeared malformed in all
but two dogs (Fig. 1). A median of two anagen bulbs was
noted in biopsies from each dog (range 08). Biopsies
from 11 dogs contained only small anagen bulbs (Fig. 2),
one dog had a large anagen bulb and two dogs had minimal numbers of both large and small anagen bulbs. Sebaceous and apocrine glands were present in all tissues. All
but two had pigmentation of the epidermis. Prominent
aggregates of melanin were present in follicular keratin
from tissue samples from 10 dogs and within basal cells
of the epidermis and hair bulbs in three dogs (two black
and one orange). Three dogs had pigment aggregates in
the sebaceous gland ducts.
After 3 months on melatonin, all skin samples had
hyperkeratosis, follicular keratosis and an epidermis of
12 cell layers thick (Table 1). Flame follicles were noted
in biopsies from 10 of 15 dogs and malformed follicles were

Table 1. Histopathological characteristics of follicular arrest in 15


Pomeranian dogs before and after 3 months of melatonin
Histopathology

No. (before)

No. (after)

Hyperkeratosis
Follicular keratosis
Prominent tricholemmal keratin
Thin epidermis
Malformed follicles
Epidermal pigmentation
Melanin aggregates
Follicular keratin
Basal cells
Sebaceous gland ducts

15
15
13
14
13
13

15
15
10
15
12
7

10
3
3

8
4
3

2006 The Authors. Journal compilation 2006 European Society of Veterinary Dermatology.

253

Frank et al.

Figure 1. Photomicrograph of skin of a dog with follicular arrest. Note


the hyperkeratosis, follicular keratosis, excess trichilemmal keratinization and malformed follicles. H&E. Bar = 500 m.

Figure 4. Photomicrograph of oestrogen receptor- staining. Greater


than 50% of basal cell nuclei within sebaceous glands have moderate
to marked staining intensity. Nuclear staining of follicular epithelium of
telogen follicle is present. There is no staining of nuclei within follicular epithelium of anagen follicles (arrows). Bar = 200 m.

Table 2. Median oestrogen receptor- staining intensity in small hair


bulbs and follicular epithelium of telogen hairs

Site
Small hair bulb
Follicular epithelium

Staining intensity
Baseline

Staining intensity
After 3 months
melatonin

++

+++

++

+++

2
2

3
2

2
2

2
1

3
2

Scale: none = 0; < 25% = 1; 2550% = 2; > 50% = 3.


+ = mild staining intensity; ++ = moderate staining intensity;
+++ = marked staining intensity.

Figure 2. Photomicrograph of skin of a dog with follicular arrest. Small


anagen bulbs are present in the deep dermis (arrows). H&E. Bar = 500 m.

Figure 3. Photomicrograph of skin of a dog with mild hair regrowth


after 3 months of oral melatonin. Many large anagen bulbs are
present in the deep dermis. H&E. Bar = 1.0 m.

observed in all but three dogs. A median of two anagen


bulbs were noted in biopsies from each dog; however, the
range was 0 15. Biopsies from five dogs had only small
anagen bulbs, three dogs had only large anagen bulbs
254

(Fig. 3) and one dog had both small and large anagen
bulbs. The three dogs with the large anagen bulbs had an
obvious increase (from 3 to 15, 0 to 6, and 2 to 11) in the
number of anagen follicles present. Owners of two of
these dogs had reported mild to moderate hair regrowth.
Biopsies from six dogs, including these two, showed
increased thickness of the follicular epithelium, suggestive of an increase in anagen hairs, although an increase in
number of anagen bulbs was not observed in four of these
dogs. Of these four dogs, only one owner reported moderate
hair regrowth. Eight of the 13 dogs with prominent epidermal pigmentation prior to melatonin treatment showed only
mild pigmentation within the epidermis after 3 months on
melatonin. Four dogs had aggregates of melanin within
the basal cells of the epidermis and hair bulbs; three dogs
had pigment aggregates in the sebaceous gland ducts.
Immunohistochemical evaluation
The sebaceous glands in all samples had moderate to
marked staining of greater than 50% of the basal cell
nuclei in all tissue samples (Fig. 4). Therefore, this helped
serve as an internal control. There was no staining of
nuclei within the epidermis, apocrine glands or dermal
fibroblasts in any tissue samples.
All of the small hair bulbs had mild to marked staining
intensity of greater than 50% of the nuclei before or after
melatonin treatment (Table 2) (Figs 5 and 6). Most of the

2006 The Authors. Journal compilation 2006 European Society of Veterinary Dermatology.

Oestrogen receptors in dogs

Figure 5. Photomicrograph of oestrogen receptor- staining. Nuclear


staining of follicular epithelium of telogen follicle is present. There is
marked staining of nuclei within small bulbs (arrows). Bar = 200 m.

Figure 7. Photomicrograph of oestrogen receptor- staining from


same dog as Fig. 3. Nuclei of anagen bulbs lack obvious staining.
There is moderate staining intensity within the dermal papilla of
several nuclei within the hair follicle on the left. Bar = 200 m.

Figure 6. Photomicrograph of oestrogen receptor- staining. Note


marked staining of nuclei within small bulbs (arrows). Positive staining
of sebaceous gland is evident (arrow head). Bar = 100 m.

large anagen hair bulbs had no oestrogen receptor staining


of the nuclei (Fig. 7). Oestrogen receptor staining was
lacking in virtually all large anagen hair bulbs, the exception
being moderate staining was noted in a few nuclei in two
anagen bulbs. The nuclei of the follicular epithelium surrounding telogen hairs in all samples had mild to marked
staining intensity of oestrogen receptors (Table 2; Figs 4
and 5). In many cases the more prominent staining
appeared to be around flame follicles (Fig. 8). Some of the
follicular epithelium was so thin that individual cell nuclei

Figure 8. Photomicrograph of oestrogen receptor- staining.


Moderate to marked staining intensity of nuclei within follicular
epithelium of telogen hair (flame follicle). Bar = 100 m.

could not be readily identified. No nuclear staining was


evident in the follicular epithelium of anagen hairs (Fig. 4).

Discussion
Hair cycle arrest (alopecia X) is described as occurring in
young adult dogs between 1 and 3 years of age.1,2 The
median age of onset of hair loss in the present study was
1 year. As in previous reports, both males and females are
affected with this condition.1,2 There were more males

2006 The Authors. Journal compilation 2006 European Society of Veterinary Dermatology.

255

Frank et al.

than females entered into the present study but intact


females were excluded. As in previous reports, both intact
and neutered dogs were affected.1,2 Neutering is considered the first-line approach for treating dogs with hair
cycle arrest.1,2 Seven dogs were neutered after the onset
of hair loss but failed to respond to surgery. Hair cycle
arrest does not appear to be restricted to coat colour since
orange, cream, black and blue colours were affected.
While more dogs with orange coat colour were entered
into the study, red, orange, and cream are the most popular coat colours of the Pomeranian breed.17 There was evidence of hair regrowth in 40% of the dogs entered into
this study. As this was an uncontrolled study, hair
regrowth cannot be directly attributed to the melatonin.
Spontaneous regrowth of hair in dogs with hair cycle
arrest, while uncommon, has been reported.18
The prominent histopathological features of hair cycle
arrest in the present study were epidermal and follicular
hyperkeratosis, prominent trichilemmal keratinization
(flame follicles), epidermal thickness of 1 2 cell layers and
lack of anagen hairs. These features are similar to what is
described in the literature and are often termed an endocrine pattern of alopecia.1 This has also been termed an
atrophic pattern19 or, more recently, a noninflammatory
pattern of alopecia20 or hair cycle abnormality.21 In addition, malformed follicles were a common finding, similar
to that described for cyclic flank alopecia.22 Cyclic flank
alopecia is an unlikely diagnosis for the Pomeranian dogs
in the present study because of the signalment, clinical
distribution of lesions and lack of cyclic activity. Abnormal
telogen follicles may be associated with a prolonged telogen state and be a nonspecific finding.21 Melanin clumping was an occasional finding in Pomeranian dogs with hair
cycle arrest, similar to that described for colour dilution
alopecia.23 These dogs were cream or orange in colour;
therefore, the melanin clumping was likely related to a
dilution gene. It is unlikely that this was the cause of the
alopecia since their colouration was generalized, but the
alopecia was not, and perifollicular melanosis was not
observed. Sebaceous gland melanosis was also noted in
a total of five dogs (one dog having this both before and
after melatonin), similar to what has been described in
dogs with endocrine skin disease or follicular dysplasia.24
Given the overlap in observed epidermal and adnexal histological features, it is apparent that histopathology alone
cannot diagnose hair cycle arrest, but merely support it by
confirming a noninflammatory cause of the alopecia.
Thirteen of 15 dogs had increased pigmentation of the
skin at the time of the first biopsy. Eight dogs showed
a decrease in pigmentation after 3 months of melatonin,
of which three dogs had no concurrent evidence of hair
regrowth. This finding supports the potential role of
oestrogen receptors in the pathogenesis, since epidermal
melanocytes are oestrogen responsive, and suggests that
melatonin may be altering the pigmentation through
oestrogen receptor blockade.25
The only histopathological changes associated with hair
regrowth were increased number of anagen follicles and
an increased thickness of the follicular epithelium associated with the anagen follicles. Interestingly, tissue samples from three dogs whose owners did not report any
evidence of hair regrowth had increased thickness of the
256

follicular epithelium. These owners were contacted to see


if hair regrowth had occurred after the completion of the
study. All owners said it had not. A possible explanation for
the discrepancy is that veterinarians in these cases biopsied sites with evidence of hair regrowth even though the
owners did not perceive any improvement in their pets
coats.
Oestrogen receptor- staining was noted in the nuclei
of sebaceous glands, follicular epithelium of telogen hair
follicles and small hair bulbs. Neither large anagen hair
bulbs nor follicular epithelium stained positive for the oestrogen receptor. It is possible that activation of the oestrogen receptors in these small bulbs is preventing the
initiation of anagen. Similar small hair bulbs are described
for postclipping alopecia.26 Further work is needed to
investigate the oestrogen receptor staining intensity of
other alopecic conditions. Because biopsy location was
not noted, no inference with regard to oestrogen receptor
distributions could be made.
There was no oestrogen receptor staining of nuclei
within the epidermis, apocrine glands or dermal fibroblasts in any tissue sample. This is in contrast to a previous
study that showed oestrogen-receptor-positive cells in the
epidermis, outer root sheath and dermal papilla of normal
dogs.12 In humans, oestrogen receptor- is expressed in
the epidermis, whereas oestrogen receptor- is not.27 In
addition, both oestrogen receptors are distributed in the
sebaceous glands, and only oestrogen receptor- is
widely expressed in the hair follicle. There is no nuclear
staining for oestrogen receptor- in the dermal papilla cells
and only weak cytoplasmic staining in some of the epithelial matrix cells of anagen hairs. The present study used an
immunohistochemical-stain-specific for oestrogen receptor-. The study by Bratka-Robia12 did not specify which
oestrogen receptor was detected in their study; however,
it is likely that they were detecting oestrogen receptor-
either exclusively or in addition to oestrogen receptor-. It
appears that, at least in mice, oestrogen receptor- is the
one primarily involved in the regulation of the hair follicle
cycle28 and that oestrogen receptor- may help regulate
oestrogen receptor-.10
There was no decrease in oestrogen receptor staining in
dogs receiving melatonin with mild to moderate hair
regrowth. The mechanism by which melatonin blocks the
oestrogen receptors is still being elucidated, and seemingly conflicting results have been reported. Melatonin in
combination with epidermal growth factor resulted in
trans-activation of the oestrogen receptor in a breast cancer cell line.29 This contradicts the known clinical effect of
melatonin. The authors theorized that the trans-activation
does not result in full transcription of the oestrogenresponsive genes, but may make the receptor refractory
to activation by estradiol. Other work has suggested that
melatonin may destabilize rather than down regulate the
oestrogen receptor.13 Most recently it has been proposed
that melatonin does not bind to oestrogen receptor-,
but instead acts as a biological modifier to affect transcriptional activity.14
In conclusion, hair regrowth in dogs with hair cycle
arrest (alopecia X) was not associated with a change in the
number or distribution of oestrogen receptor- within
either the anagen or the telogen stage follicle. Further

2006 The Authors. Journal compilation 2006 European Society of Veterinary Dermatology.

Oestrogen receptors in dogs

work with a more potent oestrogen receptor blocker to


investigate the role of oestrogen receptor- in the regulation of the hair cycle in dogs with hair cycle arrest may
yield more definitive insight into the mechanisms responsible for this condition and allow the application of more
effective treatment strategies.

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Rsum Cette tude rapporte ltude immunohistochimique des rcepteurs aux oestrognes chez les
chiens prsentant un arrt du cycle folliculaire (Alopcie X). Le but tait de dterminer si la croissance des
poils de chiens traits avec la mlatonine tait associe une diminution des rcepteurs folliculaires aux
oestrognes. Quinze Pomeranians (aucune femelle non strilise) prsentant un arrt du cycle folliculaire
ont t enrls. Deux biopsies ont t obtenues sur des zones alopciques du tronc avant et aprs trois
mois de traitement avec la mlatonine. Les sections ont t examines et une tude immunohistochimique
a t ralise pour dmontrer le rcepteur alpha aux oestrognes. Les modifications histopathologiques
regroupaient une hyperkratose, une kratose folliculaire, une kratinisation trichilemmale augmente
(follicules en flammes), un piderme fin, des bulbes anagnes rares une pigmentation pidermique et des
aggrgats de mlanine dans la kratine folliculaire. Des aggrgats de mlanine dans les cellules basales et
les poils taient occasionnellement retrouvs. Aprs trois mois, quarante pour cent (6 chiens) prsentaient
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une repousse modre des poils. Les biopsies des 6 chiens prsentaient une augmentation du nombre de
follicules en phase anagne, et huit chiens avaient une diminution de la pigmentation pidermique. Un
marquage modr marqu du rcepteur aux oestrognes a t not dans toutes les cellules basales des
glandes sbaces, tous les petits bulbes folliculaires, et lpithlium folliculaire des poils en phase tlogne.
Aucun marquage na t not dans lpidermien les glandes apocrines ou les fibroblastes dermiques. Les
follicules anagnes de grande taille ne prsentaient pas de marquage. La repousse pilaire ntait pas associe une modification du marquage du rcepteur aux oestrognes.
Resumen El papel de los receptores de estrgeno en perros con arresto del ciclo de crecimiento folicular
(alopecia X) se investig mediante tcnicas de inmunohistoqumica. El objetivo de este estudio fue determinar si el crecimiento del pelo en perros con arresto del ciclo folicular y tratados con melatonina estaba
asociado a una disminucin en la cantidad de receptores de estrgeno. Quince perros de raza Pomeranian
(excluyendo hembras no esterilizadas) con arresto del ciclo folicular fueron includos en el estudio. Se obtuvieron dos biopsias de las reas con alopecia del tronco antes y despus de tres meses de tratamiento con
melatonina. Se examinaron los tejidos tras tincin con hematoxilina y eosina, mientras que los receptores
de estrgeno se demostraron con una tcnica immunohistoqumica. Los hallazgos histopatolgicos ms frecuentes incluyeron hiperqueratosis, queratosis folicular, excesiva queratinizacin tricolemal (foliculos en
llama), delgadez de la epidermis, reducido nmero de bulbos de pequeo tamao en fase anagnica, pigmentacin de la epidermis y agregados de melanina en la queratina folicular. Los agregados de melanina
en clulas basales y en los pelos fueron un hallazgo ocasional. Despus de tres meses, un 40% (seis perros)
presentaron crecimiento del pelo de modesto a moderado. Las biopsias de seis perros presentaron evidencia histolgica de un aumento en los pelos en fase anagnica y ocho perros presentaron disminucin de la
pigmentacin epidrmica. Una intensidad de tincin de moderada a marcada se observ para el receptor
de estrgeno alfa en todas las clulas basales de las glndulas sebceas, en todos los bulbos pilosos de
pequeo tamao y en el epitelio folicular de los pelos en fase telognica. No se observ tincin nuclear para
el receptor de estrgeno alfa en las clulas epidrmicas, glndulas apocrinas o fibroblastos de la dermis.
Los bulbos pilosos en fase anagnica de tamao grande presentaron tincin mnima o nula.para el receptor
de estrgeno. El crecimiento del pelo tras el tratamiento no estuvo asociado con un cambio en la tincin
para el receptor de estrgeno.
Zusammenfassung Die Rolle von strogenrezeptoren bei Hunden, deren Haarzyklus sich in einer Ruhephase befand (Alopezia X) wurde mittels Immunhistochemie untersucht. Das Ziel dieser Studie war es festzustellen, ob das Nachwachsen der Haare bei Hunden mit Haaren in einer follikulren Ruhephase, die mit
Melatonin behandelt wurden, mit einer Abnahme der follikulren strogenrezeptoren zusammenhngt.
Fnfzehn Pomaranier (mit Ausnahme von unkastrierten Hndinnen), deren Haarzyklus sich in einer Ruhephase befand, wurden verwendet. Zwei Biopsien wurden von den haarlosen Stellen am Rumpf vor und nach
einer 3 monatelangen Behandlung mit Melatonin entnommen. Mit H&E gefrbtes Gewebe wurde untersucht und strogenrezeptor alpha immunhistochemisch dargestellt. Typische histopathologische Befunde
waren Hyperkeratose, follikulre Keratose, exzessive trichilemmale Keratinisierung (Flammenfollikel),
dnne Epidermis, wenige kleine anagene Haarblge, epidermale Pigmentation, und Melaninaggregate im
follikulren Keratin. In den Basalzellen und in Haaren wurden Melaninaggregate gelegentlich gefunden.
Nach 3 Monaten zeigten vierzig Prozent der Hunde (n = 6) geringen bis moderaten Nachwuchs der Haare.
Die Biopsien von 6 Hunden zeigten histologisch eine Zunahme an anagenen Haaren und acht Hunde hatten
eine Abnahme an epidermaler Pigmentation. Mige bis deutliche Frbungsintensitt des strogenrezeptor
alpha wurde in allen Basalzellen der Talgdrsen, in allen kleinen Haarblgen und im follikulren Epithel der
telogenen Haare festgestellt. Die Zellkerne in der Epidermis, den apokrinen Drsen oder den dermalen
Fibroblasten zeigten keine Frbung des strogenrezeptor alpha. Groe anagene Haarblge zeigten minimale oder keine Frbung des strogenrezeptor alpha. Das Nachwachsen der Haare stand nicht im Zusammenhang mit einer Vernderung der Frbung des strogenrezeptor alpha.

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