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Immunohistochemical localization of the high-affinity NGF receptor (gp 140-trkA) in the adult human dorsal root and sympathetic ganglia and in the nerves and sensory corpuscles supplying digital skin.

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THE ANATOMICAL RECORD 240:579-588 (1994)
Immunohistochemical Localization of the High-Affinity
NGF Receptor (gpl40-trkA) in the Adult Human Dorsal Root
and Sympathetic Ganglia and in the Nerves and Sensory
Corpuscles Supplying Digital Skin
J.A. VEGA, E. VAZQUEZ, F.J. NAVES, M.E. DEL VALLE, B. CALZADA,
AND J.J. REPRESA
Departamento de Morfologia y Biologia Celular, Uniuersidad de Ouiedo, Ouiedo, Spain
ABSTRACT
Background: Nerve growth factor (NGF)is produced in target tissues of sympathetic and neural-crest derived sensory neurons, including skin, to provide them trophic support. The biological effects of NGF
on responsive cells are mediated by specific high-affinity receptors. Recently, a protein tyrosine kinase of = 140 kDa molecular weight, encoded by
the proto-oncogene trkA, has been identified as the high-affinity NGF receptor (gpl40-trkA).The present work was undertaken to study the localization of gpl40-trkA-like immunoreactivity (IR) in human peripheral ganglia (sympathetic and dorsal root ganglia), and in glabrous skin.
Methods: Lumbar dorsal root ganglia, para- and prevertebral sympathetic ganglia, and digital glabrous skin were studied immunohistochemically using a rabbit anti-gpl40-trkA polyclonal antibody. In order to accurately establish the localization of gpl40-trkA IR, the neurofilament
proteins and S-100 protein were studied in parallel in: (1)sensory and sympathetic ganglia, to label neuron cell bodies and satellite or supporting
cells, respectively; ( 2 ) human skin, to label axons, Schwann and related
cells within nerves and sensory corpuscles. Moreover, a quantitative study
(neuron size, intensity of immunostaining) was carried out on sympathetic
and dorsal root ganglia neuron cell bodies.
Results: A specific gpl40-trkA-like IR was found in: (1)a subpopulation
(65%)of primary sensory neuron cell bodies, including most of the largesized ones but also small- and intermediate-sized ones; (2) most of sympathetic neuron cell bodies (82%);(3)the perineurial cell, Schwann cells, and
large axons of the nerve trunks supplying digital skin; (4) the lamellar cells
of Meissner corpuscles; (5) the central axon, inner-core, outer-core, and
capsule of Pacinian corpuscles. In addition, the occurrence of gpl40-trkAlike IR was observed in some non-nervous tissues of the skin, including epidermis (mainly in the basal layer), sweat glands, and arterial blood vessels.
Conclusions: Present results provide evidence for the localization of
gpl40-trkA-like IR in: (1)nerve cells which are known to be NGF-responsive, and (2) non-nervous cutaneous tissues which are innervated by NGFdependent peripheral neurons. These findings suggest that, in addition to
the well-established role of NGF on sensory and sympathetic neurons, this
neurotrophin may be able to regulate some other functions on non-nervous
cells which are targets for NGF-dependent peripheral neurons.
0 1994 Wiley-Liss, Inc.
Key words: Nerve growth factor receptors, gpl40-trkA, Dorsal root ganglia, Sympathetic ganglia, Cutaneous sensory corpuscles,
Skin, Immunohistochemistry, Man
Nerve growth factor (NGF) is produced in target
Of sympathetic and neural-crest derived sensory
neurons,
skin, to provide them trophic
port (Levi-Montalcini, 1987). In addition, there is evi0 1994 WILEY-LISS, INC.
Received November 10, 1993; accepted June 21, 1994,
Address reprint requests to Jose A. Vega, Departemento de Morfologia y Biologia Celular, Facultad de Medicina, Univ. de Oviedo, Ci
Julian Claveria, sin, E-33006 Oviedo, Spain.
580
J.A. VEGA ET AL.
dence for NGF-mediated paracrine effects in the skin
itself (Yaar et al., 1991).
NGF binds two classes of receptors: the high-affinity
receptors (Kd = lop1' MI and the low-affinity receptors
(Kd = lo-' M) (see Bothwell, 1989). The first one is a
protein tyrosine kinase of = 140 kDa of molecular
weight encoded by the proto-oncogene trkA (Kaplan et
al., 1991; Barbacid et al., 1991; Ross, 1991; Meakin and
Shooter, 1992; Chao, 1993). The low-affinity one is a
glycoprotein with a molecular weight of = 75 kDa (see
Bothwell, 19891, which is able to bind all different
members of the NGF family of neurotrophins (Rodriguez-Tebar et al., 1990). The high- and low-affinity
NGF receptors are referred to here a s gpl40-trkA and
p75, respectively.
The distribution of p75 IR in the adult dorsal root and
sympathetic ganglia and their targets has been widely
studied in avian and mammalian species, including
man (Chesa et al., 1988; Sobue et al., 1989; Wyatt et al.,
1990). Furthermore, it has been reported that in the
cutaneous targets, both nerves and sensory corpuscles
express p75 IR (Byers, 1990; Ribeiro da Silva et al.,
1991; Yamamoto et al., 1992; Vega et al., 1992, 1993).
On the contrary, studies on the localization of gp140trkA in the peripheral nervous system are scarce, specially in adult human tissues. Using in situ hybridization analysis, trkA mRNA has been found in the
developing and adult sympathetic and dorsal root ganglia of rat, as well a s in these ganglia of developing
human specimens (Martin-Zanca et al., 1990; Verge et
al., 1992; Wetmore et al., 1992; Ernfors et al., 1993).
However, no data are yet available on trkA mRNA neither in the adult human sensory and sympathetic neurons, nor in their cutaneous targets. Furthermore, the
distribution of the trkA-encoded protein, namely gp140trkA, has not been studied in these locations; which is
a n important issue, since is well established that mRNA
levels do not accurately reflect the protein turnover.
In the present study we addressed the localization of
gpl40-trkA IR in the adult human dorsal root ganglia,
sympathetic ganglia, peripheral nerves, and digital
glabrous skin. To ascertain what cell types displayed
gpl40-trkA-like IR, neurofilament proteins (NFP) and
S-100 protein (SlOOP)were studied in parallel, to label
neuron cells bodies, and their peripheral processes
(Lawson and Waddell, 1991), as well a s satellite cells
and Schwann cells, respectively (Vega et al., 1991).The
distribution of both antigens in human sensory corpuscles has been described in previous reports (Haro et al.,
1991; Vega et al., 1993).
MATERIALS AND METHODS
Human lumbar dorsal root and sympathetic ganglia,
as well as samples of digital skin, were obtained from
adult subjects (age range 21 to 56 years). The lumbar
dorsal root ganglia (n = 11)were obtained from 5 subjects who died by traffic or industrial accidents, during
the process of removing the organs for transplantation.
The sympathetic ganglia (n = 13) were obtained from
6 subjects during surgery; 10 pieces were lumbar paravertebral, and 3 were prevertebral. The samples of digital skin (n = 8) were obtained from 6 subjects who
suffered traumatic amputation of one or more hand
fingers and collected within 6 h after lesion. All pieces
were dissected, washed in a cold 0.9% saline solution,
fixed for 24-48 h in 10% buffered formaldehyde (pH
7.41, dehydrated, and routinely embedded in paraffin.
lmmunohistochemistry
Sections 10 pm thick were obtained, mounted on gelatin-coated slides, and processed for indirect peroxidase-antiperoxidase immunohistochemistry a s follows.
Deparaffined and rehydrated sections were washed in
0.05 M Tris-HC1 (TBS, pH 7.4) containing 1%Triton
X-100 and 1%bovine serum albumin. Thereafter, the
endogenous peroxidase activity and the non-specific
binding were blocked. Then, sections were incubated
overnight in a humid chamber at 4°C with a rabbit
polyclonal antibody against the human gpl40-trk
(Santa Cruz Biotechnology, Santa Cruz, CA) diluted
1:lOO in PBS 0.1 M, pH 7.4. This anti-trk antibody (also
known a s trkA) is a n affinity-purified rabbit polyclonal
antibody raised against a peptide corresponding to the
residues 754-790 mapping within the carboxyl terminal domain of the predicted human gp140 encoded protein. This antibody is specific for trkA, hence, shows no
trkB nor trkC cross-reactivity. Representative sections
of dorsal root and sympathetic ganglia and skin were
incubated with mouse monoclonal antibodies anti-NFP
200 kDa (clone NE14; Boehringer-Mannhein, Germany), or anti-S100P (clone 15E2E2;Boehringer-Mannhein) both diluted to 5 pgiml.
After incubation with the primary antibodies the
sections were washed in PBS and incubated for 1 h a t
room temperature with peroxidase-labelled sheep antirabbit IgG (Boehringer-Mannhein; diluted to 5 U/ml)
or sheep anti-mouse IgG (Amersham, UK; diluted
1:lOO).Finally, the sections were washed twice in PBS
and incubated with 0.05% 3.3'-diaminobenzidinei
0.01% H202for 5-10 min.
For control purposes, incubation with the primary
antibody was omitted or a non-immune rabbit or mouse
IgG were used instead of the primary antibody. Sections were examined and photographed in a light microscope.
Quantitative Studies
The neuron size, as well as the intensity of gp140trkA-like immunostaining developed in the nerve cell
body profiles of both sympathetic and dorsal root ganglia were evaluated using a n automatic image analysis
system (MIP, Servicio de Analisis de Imagenes, University of Oviedo) as described early (Dubovy et al.,
1990). Briefly, measurements were made on 3 sections
of 5 sympathetic ganglia and of 5 dorsal root ganglia.
Sections were 100 pm apart to avoid counting the same
neuron twice. Five randomly selected fields per section
were evaluated, and the number of analyzed neurons
was 20-35 per section (60-105 neurons per ganglion).
The following measurements were made: (1) neuron
size, evaluating the mean diameter (values are expressed in pm) of nerve cell body profiles with a n apparent nucleus. Neurons were divided into three
groups: small- (<25 pm), intermediate- (26-50 pm),
and large-sized ( > 5 1 pm); results are expressed a s the
percentage of labelled neurons in each established subclass. ( 2 ) intensity of immunostaining, expressing the
values in arbitrary units of grey-levels ranging from 1
(black) to 256 (white). Neurons were divided into four
groups (64 grey-level units width each), referred here
gpl40-trkA IN THE HUMAN NERVOUS SYSTEM
581
Fig. 1. Sections of dorsal root ganglia showing distribution of S-100
protein IR (A), gpl4O-trkA-like IR (B,D,E), and neurofilament proteins IR (C). In control sections (F) no positive IR was observed. As
can be shown gpl40-trkA-like IR labels primary sensory neuron cell
bodies and satellite cells (E, arrowheads). Moreover, a strong immunostaining was present in the intraganglionic blood vessels (B and D,
arrows). Arrows in (C) denote intragangiionic axons. Scale bar in (A)
= 25 pm for A-C and F; in (D) 40 pm for D and E.
TABLE 1. Percentage of primary sensory and
sympathetic neurons displaying gpl4O-trkA-likeIR,
and intensity of gpl40-trkA-likeimmunostaining into
the different neuron-size subclasses
RESULTS
Dorsal Root Ganglia
Neuron
size
(urn)
< 25 (32%)
Dorsal root ganglia
(cell n = 342)
25-50 (56%)
>51(12%)
Sympathetic ganglia < 25 (21%)
(cell n = 428)
25-50 (69%)
>51(10%)
Intensity
of IR
% o f 5% of un- (greylevel
IR neu- labeled
rons neurons units)
13
41
11
19
55
8
19
15
1
2
14
2
1 8 6 k 16
146 2 13
9629
135 i 7
12659
131 2 6
a s strong-, high-, intermediate-, and low-immunoreactivity. Nerve cell bodies with grey-level values lower
than 240 were considered unreactive. Results are expressed as mean S.D. values within each established
neuron-size classes.
*
The distribution of the measured nerve cell profiles
into the three pre-established size-classes was 32% of
small-, 56% of intermediate-, and 12% of large-sized
cells. I n the analyzed lumbar dorsal root ganglia, specific gpl40-trhA-like IR was found in about 65% of
nerve cell bodies (Fig. 1; Table 1).Most of the largesized perikarya (11%) displayed a high or intermediate
gpl40-trhA-like IR (96 5 9 grey-level units), whereas
only a subset of the intermediate- (41%) and the smallsized (13%) resulted labelled. The intensity of IR was
146 5 13 grey-level U for the intermediate-sized neurons, and 186 ? 16 grey-level U for the small-sized
nerve cells (Fig. lB,D,E; see Table 1).Thus, the intensity of immunostaining was dependent on the neuron
size. On the other hand, although differences in the
intensity of immunostaining were encountered among
subjects, the pattern of gpl40-trkA-like distribution remained unchanged. Moreover, we have observed that
satellite cells, even that sheathing neurons lacking of
immunoreactivity, showed gp140-trhA-like IR (Fig.
lB,D,E). In comparing sections processed for gp140-
582
J.A. VEGA ET AL.
Fig. 2. Localization of S-100 protein IR (A), gpl40-trkA-like IR
(B,D,E),and neurofilament proteins IR (C) in the paravertebral lumbar sympathetic ganglia. gpl40-trkA-like IR was present in both neuronal profiles and satellite cells (D,E, arrowhead). Furthermore, blood
vessels supplying sympathetic ganglia regularly resulted labelled (B,
arrows). Under control conditions no positive IR was found and dark
areas corresponded with intraneuronal storage of lipophuscin (F).
Scale bar in (A) = 25 pm for A-C and F; in (D) 20 pm for D and E.
trkA, NFP and SlOOP (Fig. lA,B,C) the localization of
gpl40-trkA-like IR in the neuron cell bodies and satellite cells can be observed. Interestingly, blood vessels
supplying dorsal root ganglia showed the highest intensity of immunostaining (Fig. lB,D).
2B) and SlOOP IR (Fig. 2A) confirmed our observations
on the localization of gpl40-trkA-like IR.
Digital Skin
gpl40-trkA-like IR was observed in both neuronal
and non-neuronal cutaneous structures. In the deep
Sympathetic Ganglia
nerve trunks (Fig. 3B) a strong gpl40-trkA-like IR was
The distribution of the measured nerve cell profiles present in the perineurium, Schwann cells, and in a
into the three pre-established size-classes was 21% of subset of axons, primarily those of large caliber. Moresmall-, 69% of intermediate-, and 10% of large-sized over, a faint gpl40-trkA-like IR was observed in the
cells. Most of the neuronal perikarya (82%) of the pre endoneurial fibroblasts. These localizations were conand paravertebral sympathetic ganglia expressed firmed comparing adjacent sections processed for
gpl40-trkA-like IR in their cytoplasm with a rather gpl40-trkA with that for SlOOP (Fig. 3A) and NFP
homogeneous intensity of immunostaining which was (Fig. 3C) demonstration.
independent of the neuron size (Fig. 2). The immunoDifferent kinds of sensory corpuscles showing gp140reactive neurons included 19% of small-, 55%of inter- trhA-like IR were found in the digital skin. The lamelmediate-, and 8% of large-sized cells (see Table 1). lar cells of Meissner corpuscles placed in the dermal
Moreover, satellite cells displaying positive IR were papillae showed intense gpl40-trkA-like IR (Fig. 4).
also found (Fig. 2D,E), the intensity of IR being stron- Moreover, immunostaining in the central axon of
ger than that observed in the neuronal somata. On the Meissner corpuscles was not evident. With regard to
other hand, and a s i t was described for the dorsal root the Pacinian corpuscles, all the three main systems
ganglia, intraganglionic blood vessels were intensely (i.e., the central axon, inner-core, and outer-corelcaplabelled (data not shown). The comparison of the dis- sule) were immunolabelled (Fig. 5A,B). Results on Patribution for NFP IR (Fig. 2 0 , gpl4O-trkA-like IR (Fig. cini-like corpuscles and on simple lamellar corpuscles
Fig. 3. Adjacent sections of a deep cutaneous nerve incubated with
antibodies for S-100 protein (A),gpl40-trkA (B),and neurofilament
proteins (C).The gpl40-trkA-like IR was found labelling perineurium
(p), Schwann cells and axons (arrows). The endoneurial fibroblasts
also were labelled. Scale bar = 25 pm.
Fig. 4. Meissner’s corpuscles displaying gpl4O-trkA-like IR. The occurrence of IR within central axons cannot be assured, while it was
evident in the lamellar cells. e: epidermis. Scale bar = 25 Fm.
Fig. 5. Localization of gpl4O-trkA-like IR in Pacinian corpuscles
(A,B) and Pacini-like corpuscles ( C ) . All the corpuscular structures
had immunolabelling with the intensity of IR being stronger in the
inner-core lamellae. Inset in (B) corresponds to an approximate section of the Pacinian corpuscle processed for detection of neurofilament
proteins. Arrows indicate the axons; c: capsule; ic or arrowheads: inner-core; oc: outer-core. Scale bar = 25 pm
584
J.A. VEGA ET AL.
were similar, with a prominent gpl40-trkA-like IR
within the central axon and inner-core lamellar (Fig.
5C). Localization of gpl4O-trkA-like IR on sensory corpuscles was confirmed by comparing sections processed
for gpl40-trkA-like detection with that for NFP (inset
in Fig. 5B) and SlOOP (data not shown; see Vega et al.,
1993).
Regarding non-nervous tissues, we observed the presence of gpl40-trkA-like IR in the epidermis, sweat
glands, and blood vessels. In the epidermis, the stronger
IR was in the basal layer, while the suprabasal layers
showed a very faint gpl40-trkA-like IR (Fig. 6).
Whether or not Merkel cells show IR cannot be ensured
since no specific techniques to label Merkel cells were
done. Furthermore, gpl40-trkA-like IR cells showing a
dendritic-like morphology were found at the level of the
suprabasal layer (Fig. 6A) or within the basal layer
(Fig. 6A,B). They could be identified as Langerhans
cells and/or melanocytes. In addition to the epidermis,
other epithelial structures within the dermis displayed
gpl40-trkA-like IR. We identified them as sweat glands
(Fig. 7), and the IR intensity was stronger in the ductal
part than in the acinar one. At the light microscope level
it is not possible to ascertain whether IR labelled myoepithelial cells.
Finally, gpl40-trkA-like IR was identified in the
walls of both large and small arteries (Fig. 81, but not
in the veins (Fig. 8B). The labelling was mainly found
in the smooth muscle cells of the tunica media, and
occasionally also in the adventitia a s nerve-like profiles. Apparently, no positive immunostaining was
present in the endothelial cells.
Under control conditions, where the primary antibody was omitted or a non-immune rabbit or mouse
IgG were used instead of the primary antibody, no specific IR was observed for any of the assessed antibodies
(Figs. l F , 2F, and 6C).
DISCUSSION
This paper reports for the first time the distribution
of gpl40-trkA-like IR in the adult human dorsal root
and sympathetic ganglia, in the peripheral nerves, as
well a s in their cutaneous targets. These included sensory corpuscles, epidermis, sweat glands, and blood vessels. The exact nature of the gpl40-trkA-like IR cells in
the nervous structures was established comparing the
pattern of gpl40-trkA-like IR with that of markers for
neurons (NFP), or satellite cells (Sloop) whose distribution is well known in the peripheral ganglia and
nerves (Lawson and Waddell, 1991; Vega et al., 1991) as
well a s in the sensory corpuscles (Vega et al., 1993).
The neurons of the sympathetic and dorsal root ganglia (see Levi-Montalcini and Calissano, 1986; Barde,
1989; Verge et al., 19931, as well as neuronal subpopulations of some cranial sensory ganglia, are under
NGF control during development (Davies and Lindsay,
1985; Represa and Bernd, 1989). These neurons express both low- and high-affinity NGF receptors during
development (Bernd and Represa, 1989; Wyatt et al.,
1990) and also in post-natal periods (Ernfors et al.,
1993). However, whether or not NGF is essential for
maintenance of mature peripheral neurons is unclear
(Lindsay, 1988; Gold et al., 1991). Data concerning the
presence of NGF receptors in adult human sympathetic
and dorsal root ganglia are restricted to the low-affin-
ity receptors, thus the p75 protein (Sobue e t al., 1989).
Present results demonstrate that most of postganglionic sympathetic neurons (82%) and a wide subset of
sensory dorsal root ganglia neurons (65%) express
gpl40-trkA-like IR, thus suggesting a n active role of
NGF in these mature and differentiated nerve cells
(Lindsay, 1988).
We have constantly observed gpl40-trkA-like IR in
nerve trunks supplying digital skin, labelling perineurial cells, endoneurial fibroblasts, Schwann cells, and
large axons. The pattern of gpl40-trkA-like IR in these
localizations matches the pattern of distribution of p75
IR in human nerve trunks (see Vega et al., 1992). However, our findings are in disagreement with those reported by Yamamoto et al. (1993) who could not observe
expression of gpl40-trkA mRNA in cultured Schwann
cells. Specific or technical differences may account for
these discrepancies (i.e., levels of mRNA might be undetectable while levels of protein might not be).
Both Meissner and Pacinian corpuscles expressed
gpl4O-trkA-like IR with a n identical localization to that
reported for p75 (Vega et al., 1993). At the light microscope level, nevertheless, whether immunolabelling occurs within the cytoplasm or in the membrane of lamellar cells cannot be ensured. Results obtained in sensory
corpuscles may be considered together with that on
nerve trunks, due to the fact that the axon, lamellar
cells or inner-core and outer-corelcapsule of sensory corpuscles are continuous with the axon, Schwann cells,
and perineurial cells, respectively, of the nerve fibers
(Malinovsky, 1986; Munger and Ide, 1988). However, i t
might also be possible that NGF receptors present in
sensory corpuscles play a specific and distinct role in
these structures (Ritter et al., 1991; Lewin e t al., 1992;
Lewin and Mendell, 1993), although this issue remain
controversial (Diamond e t al., 1992a,b).
As above described, gpl40-trkA-like IR was mainly
found in intermediate- and large-sized primary sensory
neurons and, probably, in their parent axons in cutaneous nerves. Interestingly, the axons entering Meissner and Pacinian corpuscles represent the specialized
ending of A@and A6 nerve fibers (see Dalsgaard, 1988;
Fyffe, 1992) originated from large- and medium-sized
sensory neurons (Lawson and Waddell, 1991). Taken
together these observations suggest that expression of
gpl40-trkA could be related with non-nociceptive
mechanoreception.
Several studies have suggested a n involvement of
NGF in the control of different functions in non-neuronal tissues, other than the classical neurotrophic effects. These tissues include keratinocytes, dendritic
cells, sweat glands, and blood vessels of the human
skin, since all of them express p75 IR (see Chesa et al.,
1988; Ernfors et al., 1991; Vega e t al., 1992). We have
now demonstrated the occurrence of gpl40-trkA-like IR
in the same non-nervous cutaneous structures, in a
pattern which closely resembles that of p75 protein.
However, whether gpl40-trkA-like IR in the basal keratinocytes is related with the so-called trkE (Di
Marco et al., 1993) must be clarified in future studies.
gpl40-trkA-like IR was observed in the cutaneous
blood vessels, primarily in the tunica media. No data
are so far available on this topic. However, the distribution of gpl40-trkA-like IR showed here basically
matches the localization of p75 IR in different blood
gpl40-trkA IN THE HUMAN NERVOUS SYSTEM
Fig. 6. In the epidermis most cells the basal layer were gpl40-trkAlike IR positive. Within the basal layer and In the suprabasal layers
of the epidermis, cells with a dendritic morphology and displaying
gpl40-trkA-like IR were observed (A,B, arrowheads). e: epidermis.
Scale bar = 25 pm.
Fig. 7. Sweat glands showing gpl40-trkA-like IR which was strong
in the ductal portions (arrows)and moderate in the acini. Scale bar =
25 pm.
585
Fig. 8. Large (A)and small (B)cutaneous arteries displaying gp140trkA-like IR. In the large ones IR was primarily localized in the tunica
media while in the small ones the IR occupied all wall layers. The
veins did not display IR. a: adventitia; ar: artery; tm: tunica media; v:
vein. Scale bar = 100 pm in A, and 25 pm in B.
586
J.A. VEGA ET AL.
corpuscles, as well a s for the epithelial cells forming
the sweat glands.
As can be observed in Table 2, most cells producing
NGF also express both types of NGF receptors. Theregpl40-trkANGF
like IR
p75-IR2 secretion3 fore, our results strongly suggest t h a t NGF produced
and released in human skin may (1)act on NGF-reNerve bundles
sponsive sensory and sympathetic neurons, being con+++
+++
?
Perineurium
ducted
to them throughout axonic retrograde trans++
++
?
Endoneurial fibroblasts
port; (2) have a major role in regulating functions in
+++
+++
Yes
Schwann cells
some non-neuronal cutaneous cells working in a n au++
+I No
Axons
tocrine or paracrine manner, as is suggested for keraSensory corpuscles
tinocytes and dendritic cells (Di Marco et al., 1991;
++
+ +I+
?
Capsule*
Yaar et al., 1991). In this way Schecterson and Both++
+ +I+
?
Outer-core*
well (1992) have suggested a role for NFG in neuron to
+++
+++
?
Schwann-related cells
target tissue signaling. Moreover, since non-neuronal
++
+INo
Axon
cutaneous cells displaying gpl40-trkA-like IR also exEpidermis
press p75 IR, a collaboration between both types of
+++
++
Yes
Basal keratinocytes
NGF receptors could be hypothesized for the skin. How+I+
Yes
Suprabasal layers
?
++
?
Merkel cells
ever, co-expression of both NGF receptors to mediate
+++
+++
?
Dendritic cells
biological responses of NGF in other systems remains
controversial (Glass et al., 1991; Hempstead et al.,
Dermis
Fibroblasts
1991; Ibanez et al., 1992; Lee et al., 1992). Further
Muscle cells
studies will be necessary to clarify the role of NGF in
the human skin cells which express NGF receptors,
Sweat glands
++
+++
?
Acini
and the functional and clinical relevance of the present
+++
+++
?
Excretory ducts
results remain to be elucidated.
TABLE 2. Distribution of gpl4O-trkA-likeIR
and p75 IR in human digital skin, and cells
synthesizing and secreting NGF'
-
Blood vessels
Endothelium
Muscular layer
Adventitia
-
++
-
-
+
+ +I+
No
Yes
?
'+ + + indicates strong, + + moderate, and + weak I R - indicates
no I R ? = data not done; * indicates Pacinian corpuscles.
'From Chesa et al. (1988) and Vega et al. 11992, 1993).
3From Rohrer et al. (1988), Creedon and Tuttle (19911, Di Marco et al.
(1991). Yaar et al. (1991), Thoenen et al. (19921, Yamamoto et al.
(1993).
vessels (see Del Valle e t al., 19921, including the cutaneous ones (Vega et al., 1992). Our observations suggest that vascular smooth muscle cells might be responsive to NGF. In supporting this opinion, Messina
and Bell (1991) and Zettler et al. (1991) demonstrated
that NGF may be involved in the regulation of blood
vessel pressure, probably throughout a sympathetic
way (Isaacson et al., 1990) since NGF acts as a n inhibitory neuromodulator of adrenergic transmission in the
rat mesenteric artery (Ueyama et al., 1991).
NGF is synthesized and released by different cells
present in the developing and adult human skin (see
Table 2) including keratinocytes (Rohrer e t al., 1988;
Wyatt et al., 1990, Di Marco et al., 1991; Yaar et al.,
1991), fribroblasts, Schwann cells, and smooth muscle
cells (Creedon and Tuttle, 1991; Thoenen et al., 1992;
De Vellis, 1993; Reynolds and Woolf, 1993). In this order, the tunica media of elastic arteries (MacGrogah et
al., 1992; Scarisbrick et al., 1993) contain high mRNA
levels for neurotrophins, including NGF. Although no
data are so far available on the synthesis and release of
NGF from perineurial cells and their corpuscular derivatives, a presumptive NGF production by these cells
could be assumed (see Yoshida and Gage. 1992: Thoenen et al., 1992), since they have a fibroblastic' origin
(Halata et al., 1990). The Same may apply for the
Schwann-related cells of the Meissner and Pacinian
ACKNOWLEDGMENTS
Present study was supported by grants from FISS
(91/0361) and from the University of Oviedo (DF-92/63,
DF-93 218-57, and TA-93 218-35). E.V. was recipient of
a grant fellowship from DGICYT (92/0261). Technical
assistance of Mr. J.A. Garcia-Sanchez is gratefully acknowledged.
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dorsal, supplying, nerve, high, trka, skin, sensore, sympathetic, human, 140, roots, adults, immunohistochemical, localization, digital, ganglia, ngf, corpuscles, receptov, affinity
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