вход по аккаунту


Serotonin-like immunoreactivity in Merkel cells and their afferent neurons in touch domes from the hairy skin of rats.

код для вставкиСкачать
THE ANATOMICAL RECORD 232:112-120 (1992)
Serotonin-Like lmmunoreactivity in Merkel Cells and Their Afferent
Neurons in Touch Domes From the Hairy Skin of Rats
Department of Physiology, University of Utah School of Medicine, Salt Lake City, Utah
Immunoreactivity to serotonin was observed in Merkel cells as
well as the afferent type I nerves terminating upon them in touch domes excised
from the belly skin of rats. Type I nerves were strongly immunoreactive and could
be traced through the dermis of the domal papilla. Merkel cell immunoreactivity
was sometimes seen in the entire cell, but was often localized in the Merkel cell
cytoplasm adjacent to nerve terminals and may have been in the terminals themselves. Domes were fixed by immersion in 4% paraformaldehyde-lysine-sodiumm-periodate (PLP) fixative at 4°C for 2.5-3 hours and cryoprotected in 30% sucrose
overnight. Sections were processed with the avidin-biotin complex peroxidase
(ABC), peroxidase-antiperoxidase(PAP), and indirect immunofluorescence techniques with rabbit antiserum generated against serotonin.
Merkel cells are situated in the basal epidermis of
the most tactually sensitive regions of vertebrate skin.
They are found in fish (Lane and Whitear, 1977), amphibians (Crowe and Whitear, 1978) and mammals, including humans (Merkel, 1875; Smith, 1970). Their
presence in so many classes implies they offer a selective advantage in those animals possessing them
(Simon and Hiller, 19891,but just what that role is has
yet to be verified.
In the hairy skin of rats, Merkel cells have a high
density in the upper cylindrical cuff of the external root
sheath of vibrissae. They are also in the basal epidermis of touch domes located on trunk skin or the whisker pad (Nurse et al., 1983; Nurse and Diamond, 1984).
Following depilation, domes are visible as small elevations (170-314 pm in diameter; English, 1977; Leon
and McComas, 1984) rising above the skin surface. On
the trunk of adult rats, they are spaced about 3-5 mm
apart and are associated with large tylotrich hairs
(Straile, 1960).
Almost all Merkel cells are innervated (Pasche et al.,
1990). A type I afferent fiber forms a discoid shaped
terminal on the dermal side of a Merkel cell’s surface
(Iggo and Muir, 1969; English et al., 1980). Merkel
cells characteristically contain dense cored granules
-100 nm in diameter that are polarized next to the
nerve terminals (Fig. 1).Sometimes granules fuse at
synapse-like junctions (Chen et al., 1973). Mechanical
depression of touch domes elicits a n irregular, slowly
adapting discharge in the afferent type I fibers (Iggo
and Muir, 1969).
There is physiological evidence in support of two conflicting hypotheses: either Merkel cells transduce mechanical energy into nerve impulses (Horch e t al.,
1974; Findlater et al., 1987), or type I nerve endings
are the true receptors (Diamond et al., 1988; Gottschaldt and Vahle-Hinz, 1981). In addition, Merkel
cells have been proposed to be the following: (1)paracrine regulators of the epidermis (Moll et al., 1986), (2)
targets for growing nerves (Scott et al., 1981; Diamond,
1982; Yasargil et al., 1988), (3) neuromodulators of the
nerve terminal’s mechanosensitivity, or (4) developmental “positional labelers” for the central nervous
system (Diamond et al., 1988).For any of these functions, the contents of the dense cored granules and the
stimuli causing their release are likely to be of central
Because serotonin has previously been shown to excite type I cutaneous afferent neurons in vivo (Beck
and Handwerker, 1974), the present immunohistochemical study was conducted to determine if i t might
be found in Merkel cell-axonal complexes. If present,
serotonin’s role as a neurotransmitter in this system
could be tested in a recently modified in vitro skin
preparation (Reeh, 1986).
Adult Sprague Dawley rats of both sexes were anesthetized with a n intramuscular injection of ketamine
hydrochloride (100 mgiml) mixed with acepromazine
(10 mg/ml) at 10 parts to 1, followed by 4% chloral
hydrate administered subcutaneously according to
body weight. Because of their large size, domes were
removed from the belly after the fur was clipped and
depilated (Nair).
The tissue was (1) immersed into ice-cold, freshly
prepared 4% paraformaldehyde-lysine-sodium-m-peroidate (PLP) fixative in 0.1 M phosphate-buffered saline
(PBS; pH 7.4) for 2.5-3 hours; (2) rinsed in PBS for 10
minutes, followed by 15% sucrose in PBS for 1 hour;
and (3) left in 30% sucrose in PBS overnight at 4°C.
Individual domes were oriented on their side for sectioning, snap frozen in OCT, sectioned at 5-10 km, and
Received March 22, 1991; accepted May 10, 1991.
Address reprint requests to Kathleen B. English, Ph.D., Department of Physiology, University of Utah School of Medicine, 410 Chipeta Way, Research Park, Salt Lake City, UT 84108.
Fig. 1. Electron micrograph of a Merkel cell (MI and its associated
type I nerve terminal (NT) with numerous mitochondria. The touch
dome was excised from an adult rat. The Merkel cell has dense cored
granules (g) in its cytoplasm. Spine-like processes (*) extend among
neighboring keratinocytes of the epidermis. Merkel cells are above
the epidermal basement lamina (bl) and form desmosomes (d) with
the keratinocytes. Scale bar = 0.5 pm.
mounted on chrome alum-subbed slides. The tissue was
processed for immunocytochemical staining using conventional peroxidase-antiperoxidase PAP (Sternberger, 1986), avidin-biotin complex peroxidase ABC (Hsu
et al., 1981), and indirect immunofluorescence techniques (Coons, 1958).
Sections were stained with rabbit primary antiserum
to serotonin [1:5,000 (NT102 [lyophilizedl; Eugene Tek
Int.) or 1:200-300 (AB125; Chemicon)] overnight a t
4°C. Blocking serum was 10% normal goat serum in 0.1
M PBS saline. All antibodies were diluted in 0.1 M PBS
saline plus 1% normal goat serum and 1%crystalline
grade bovine serum albumin (Sigma, A7638). A 10minute wash in 3% H,02 quenched endogenous peroxidase activity in the skin. Biotin in sebaceous glands
was bound up with a n avidin-biotin blocking step
For the ABC and PAP techniques, the secondary
antisera were biotinylated goat anti-rabbit (1:400; Vector) or goat antiserum to rabbit (1:lOO).The PAP complex was prediluted (Dakopatts). The reaction products
were visualized by the diaminobenzidine (DAB) reaction, which was closely monitored under a brightfield
microscope. DAB was freshly filtered and prepared
each run using 10 mg DAB (Sigma), 20 mls 0.1 M PBS,
pH 7.4, and 4 ~1 30% HzOz (Sigma). The secondary
antiserum for indirect immunofluorescence was goat
anti-rabbit IgG-L-Rhodamine [(1:200-1:400) Boehringer Manheiml.
The specificity of the serotonin antiserum was
checked by preabsorbing serotonin antiserum at working dilution with 5-HT bound to agarose (Sigma,
H7136) or with serotonin creatinine sulfate (Sigma).
Other controls consisted of inclusion of 2 mg/ml of polyL-lysine (Sigma #P0879) in the antiserum diluent
(Scopsi et al., 19861, omission of the primary or secondary antiserum, or substituting primary antiserum with
normal, nonimmune rabbit serum (Eugene Tek Int.).
Rat carotid body served a s positive control tissue.
Serotonin-like immunoreactivity was observed in
Merkel cells, in afferent type I neurons with Merkel
cells (Fig. 2) and in dermal mast cells. Immunoreactivity in Merkel cells was often strongest in their cytoplasm adjacent to nerve terminals or in the terminals
themselves (Figs. 3A, 4A). The reaction time for immunostaining to occur was approximately 2-5 min. Not
every Merkel cell in a given section was immunoreactive, but roughly 25% were. Type I neurons were
strongly immunoreactive (Figs. 2B, 4B,C). Immunostaining was identical for both antisera tested (NT102
Fig. 2. A Arrows point to Merkel cells immunoreactive to serotonin
in the basal epidermis of a touch dome from a n adult rat. ABC immunoperoxidase technique, interference contrast microscopy. Scale
bar = 10.0 pm. B: Arrows point to Merkel cells in the epidermis, and
the open arrow points to the type I nerve in the dermis of a touch
dome. Both are imrnunoreactive to serotonin. ABC irnmunoperoxidase technique, interference microscopy. Scale bar = 10.0 pm.
Fig. 3. A Section of a touch dome with Merkel cells (arrows) and
their nerve terminals immunofluorescently labeled against serotonin. Scale bar = 10.0 km. B: Control section of a touch dome where
normal rabbit serum was substituted for the primary antiserum. Sections receiving preabsorbed antiserum were identical in appearance.
Scale bar = 10.0 pm.
Fig. 4. A Touch dome from an adult rat. Arrows point to type I
nerve terminals immunofluorescently labeled for serotonin. Scale bar
= 10.0 pm. B,C: Open arrows point to immunofluorescent type I
nerves labeled for serotonin. The nerves rise in the dermis and terminate upon Merkel cells (arrow) in the basal epidermis of touch
domes. Scale bar = 10.0 pm.
Ilyophilizedl Eugene Tek Int. and AB125 Chemicon).
Glomus cells of the carotid body were immunoreactive
for serotonin (Gronblad et al., 1983) and thus substantiated the specificity of the antisera tested.
Control sections incubated without primary or secondary antisera failed to demonstrate any immunoreactivity, as did sections incubated with normal rabbit
serum or antiserum preabsorbed with bound serotonin
(Fig. 3B). Preabsorption with serotonin creatinine sulfate required 10-1M concentrations for immunostaining to be eliminated and was not reliably reproducible
(perhaps because the antiserum was generated against
serotonin in its conjugated form). Inclusion of po1y-Llysine in the antiserum diluent did not abolish staining
in the Merkel cells or the type I nerves, indicating the
immunoreaction was not due to nonspecific binding of
basic peptides.
In the present investigation, serotonin-like immunoreactivity was found in Merkel cells and their afferent nerves in touch domes excised from the hairy skin
of rats. This supports previous immunohistological
studies demonstrating serotonin in Merkel cells from
the eel (Zaccone, 1986) and pig (Garcia-Caballero et al.,
1989b), as well as studies employing permanganate
staining and reserpine treatment to demonstrate
monoamines in Merkel cell granules of mice (Chao and
Liang, 1975). However, it is a t variance with other less
sensitive and selective histochemical and pharmacological investigations in cats and rats (Smith and
Creech, 1967; Iggo and Muir, 1969; Hartschuh and
Grube, 1979; see also Diamond et al., 1986).
In rats, Merkel cells have been shown to contain the
opioid peptide metenkephalin (Hartschuh e t al., 1979)
and the putative neurotransmitter adenosine triphosphate (ATP; Crowe and Whitear, 1978; Nurse et al.,
1983). In various other species, Merkel cells have been
demonstrated to be immunoreactive for neuron specific
enolase, chromogranin A, vasoactive intestinal polypeptide (VIP), substance P, pancreastatin, and calcitonin gene-related peptide (Alvarez et al., 1988; Gu et
al., 1981; Hartschuh and Weihe, 1988, 1989).
It is common for biogenic amines to be colocalized
with transmitters and neuropeptides in excitatory or
receptosecretory cells (Gauweiler et al., 1988; Kanagawa et al., 1986; Forloni et al., 1987). Indeed by definition, cells of the APUD (amine precursor uptake and
decarboxylation), DNES (diffuse neuroendocrine system), or paraneuron class characteristically possess
amines and peptides as well a s marker molecules such
as neuron specific enolase and chromogranin A (Fujita,
1983; Pearse, 1986; Hartschuh and Weihe, 1988).
In general, Merkel cells of the skin, along with chromaffin cells of the adrenal medulla, neuroepithelial
cells of the lung, enterochromaffin cells of the gastrointestinal tract, and glomus cells of the carotid body
are all considered to be neuroendocrine cells, belonging
to the APUD, DNEs, paraneuron, or receptosecretory
class of cells. The presence of biogenic amines in Merkel cells might therefore be expected, but it is not a n
absolute requirement for the receptosecretory classification to stand.
The present study expands the number of species in
which serotonin has been found in Merkel cells. As
methodologies are refined, there may be more correlation between immunohistochemical profiles of Merkel
cells (see Table 1).For instance, met-enkephalin was
originally reported to only occur in Merkel cells from
rodents (guinea pigs, mice, and hamsters), but not in
those from cats, dogs, pigs, and humans, which demonstrated immunoreactivity to vasoactive intestinal polypeptide (VIP) instead (Hartschuh et al., 1984). Yet a t
the same time other investigators found met-enkephalin immunoreactivity in Merkel cells from primates
(Warner e t al., 1983). Unfortunately, there are technological difficulties with immunohistochemical staining. This is exemplified by synaptophysin reactivity
only occurring in perfusion-fixed guinea pig tissue and
not in immersion-fixed pig and human skin. Similarly,
chromogranin A is shown with bovine-generated antiserum rather than porcine-generated antiserum
(Hartschuh and Weihe, 1988).
The existence of serotonin-like immunoreactivity
both in Merkel cells and their afferent nerves in the
present study was unexpected. It was found in Merkel
cells but not afferent nerve fibers in the pig and eel
(Zaccone, 1986; Garcia-Caballero et al., 198913). However, similar observations have been made in other
APUD cells and their sensory nerve supply. For example, in the adrenal medulla, both adrenaline cells and
their afferent neurons express immunoreactivity for
enkephalins (Schultzberg et al., 1978; Pelto-Huikko et
al., 1985). Similarly, catecholaminergic primary sensory neurons terminate upon catecholamine-containing type I glomus cells in the carotid body (Katz and
Black, 1986). The physiological significance of dual localization of putative neurotransmitter substances in
APUD cells and their afferent neurons is presently unknown.
Earlier enzymatic histochemical studies of the type I
nerve endings of Merkel cells indicated acetylcholinesterase and leucine aminopeptidase were present
in rabbits, and alkaline phosphatase in cats and guinea
pigs (Winkelmann, 1977). The present investigation is
the first study showing a specific immunohistochemical reaction in the type I nerve terminals on Merkel
cells. Other investigators have found CGRP immunoreactive nerve fibers occasionally associated with Merkel cells in the footpad of rats, but these were not the
type I fibers of the Merkel cell-axonal complex (IshidaYamamoto et al., 1989).
Serotonin is known to be ultrastructurally associated
with synaptic vesicles (Sanders-Bush and Martin,
1982). Merkel cells have numerous dense cored granules, but their afferent neurons do not. There is, however, considerable experimental evidence that a large
proportion of serotonin is stored in a cytoplasmic extravesicular form (Halaris and Freedman, 1977; Lewis
and Moertel, 1978; Sanders-Bush and Martin, 1982).
Therefore, the sparcity of dense-cored granules in the
afferent nerves terminating upon Merkel cells does not
exclude the possibility of serotonin being present in
cytoplasmic stores.
The role that serotonin plays in the Merkel cell-axonal complex remains to be determined. It may be involved in any one of several biological processes. For
example, serotonin may be important in the generation
of type I responses in afferent nerves because both serotonin formation and type I response are highly de-
TABLE 1. Merkel cell immunohistochemistry
Substance P
CGRP (calcitonin gene-related peptide)
PHYPMI (peptide histidine isoleucine)
NSE (neuron specific enolase)
Cytokeratin 8,18
PGP (protein gene product 9.5)
guinea pig, mouse, hamster
adult and fetal pig
adult cat, adult mouse, adult pig
adult and fetal pig
adult pig, adult cat, adult mouse
adult cat
adult pig
dog, cat, human, pig
adult cat, mouse, pig
adult pig
adult cat, mouse, pig
adult and fetal pig
adult human and pig
fetal human
cat, rat
adult and fetal human
adult human
rabbit, pig, human
human, rabbit, pig
adult guinea pig
adult and fetal pig
adult cat, adult pig
adult and fetal pig
Chromogranin A
pendent upon POz. Type I receptors fail to respond to
mechanical stimulation in a n oxygen-depleted environment. Furthermore, Merkel cells become degranulated
when touch domes are mechanically stimulated under
hypoxic conditions (Findlater et al., 1987). On the other
hand, serotonin may be required for synthesis and storage of polypeptides (Owman e t al., 1973). Alternatively, recent research indicates bidirectional communication exists between neuroendocrine cells and the
immune system, wherein serotonergic systems inhibit
immune responses (Warren e t al., 1990; Devoino et al.,
1990). Now that serotonin has been identified in the
Merkel cell-axon complex, its function will be the focus
of future electrophysiological and neuropharmacoligical investigations.
The authors wish to express thanks to Dr. Andy
Towles of ETI, Dr. P. Burgess for his critique of the
manusmirk. and to Vicki Skelton for twine: the manuscript. Supported by NIH grants" -NS%7938 and
Alvarez, F.J., C. Cervantes, R. Villalba, I. Blasco, R. MartinezMurillo, J.M. Polak, and J. Rodrigo 1988 Immunocytochemical
analysis of calcitonin gene-related peptide and vasoactive intestinal polypeptide in Merkel cells and cutaneous free nerve endings of cats. Cell Tissue Res. 254r429-437.
Hartschuh et al. (1979)
Hartschuh et al. (1983)
Warner et al. (1983)
Hartschuh et al. (1989)
Gauweiler et al. (1988)
Hartschuh et al. (1989)
Gauweiler et al. (1988)
Alvarez et al. (1988)
Hartschuh et al. (1989)
Hartschuh et al. (1984)
Gauweiler et al. (1988)
Hartschuh et al. (1989)
Gauweiler et al. (1988)
Hartschuh et al. (1989)
Hartschuh and Weihe ( 989)
Zaccone (1986)
Garcia-Caballero et al. 1989b)
Gould et al. (1985)
Zaccone (1986)
Gu et al. (1981)
Moll et al. (1984)
Ness et al. (1987)
Ortonne and Darmon (1985)
Dalsgard et al. (1989)
Wang et al. (1990)
Garcia-Caballero et al. (1989a)
Ortonne et al. (1988)
Hartschuh and Weihe (1988)
Hartschuh et al. (1989)
Gauweiler et al. (1988)
Hartschuh and Weihe (1988)
Beck, P.W., and H.O. Handwerker 1974 Bradykinin and serotonin
effects on various types of cutaneous nerve fibres. Pfliigers Arch.
Chao, C.-F., and H.-M. Liang 1975 Ultrastructural and cytochemical
studies of Merkel cell granules in the mouse Haarscheibe. Bull.
Inst. Zool. Academia Sinica 14r71-78.
Chen, S.-Y., S. Gerson, and J. Meyer 1973 The fusion of Merkel cell
granules with a synapse-like structure. J. Invest. Dermatol. 61:
Coons, A.H. 1958 Fluorescent antibody methods. In: General Cytochemical Methods. J.F. Danielli, ed. Academic Press, New York,
pp. 399-422.
Crowe, R., and M. Whitear 1978 Quinacrine fluorescence of Merkel
cells in Xenopus laevis. Cell Tissue Res. 190:273-283.
Dalsgaard, C.-J., M. Rydh, and A. Haegerstrand 1989 Cutaneous innervation in man visualized with protein gene produce 9.5 (PGP
9.5) antibodies. Histochemistry 92:385-389.
Devoino, L., M. Cheido, G. Idova, N. Morazova, 0. Papsuevich, and G.
Chipens 1990 Low molecular weight peptides in neuroimmunomodulation. Central effect and interaction with monoamine systems. Ann. N.Y. Acad. Sci. 594t449-451.
Diamond, J. 1982 Modeling and competition in the nervous system:
Clues from the sensory innervation of skin. In: Neuronal Development, Part 111, Neuronal Specificity, Plasticity and Patterns.
A.A. Moscona and A. Monroy, eds. Academic Press, New York.
Diamond, J., M. Holmes, and C.A. Nurse 1986 Are Merkel cell-neurite reciprocal synapses involved in the initiation of tactile responses in salamander skin? J. Physiol. 376:101-120.
Diamond, J., L.R. Mills, and K.M. Mearow 1988 Evidence that the
Merkel cell is not the transducer in the mechanosensory Merkel
cell-neurite complex. Prog. Brain Res. 74:51-56.
English, K.B. 1977 Morphogenesis of Haarscheiben in rats. J. Invest.
Dermatol. 69.38-67.
English, K.B., P.R. Burgess, and D. Kavka-Van Norman 1980 Development of rat Merkel cells. J. Comp. Neurol. 194r475-496.
Findlater, G.S., E.J. Cooksey, A. Anand, A.S. Paintal, and A. Iggo
1987 The effects of hypoxia on slowly adapting type I (SAI) cutaneous mechanoreceptors in the cat and rat. Somatosens. Res. 5:
Forloni, G., R. Grzanna, R.D. Blakely, and J.T. Coyle 1987 Co-localization of n-acetyl-aspartyl-glutamate in central cholinergic, noradrenergic, and serotonergic neurons. Synapse 1t455-460.
Fujita, T. 1983 New aspects of cells secreting neuropeptides. In: Endocrinology. K. Shizume, H. Imura, and N. Shimizu, eds. Excerpta Medica, Amsterdam, pp. 35-43.
Garcia-Caballero, T., J. Cuevas, R. Gallego, E. RosCtn, J. Forteza, and
A. Beiras 1989a Synaptophysin-like immunoreactivity in the
Merkel cells of pig-snout skin. Ultrastruct. Pathol. 1355-61.
Garcia-Caballero, T., R. Gallego, E. Roson, D. Basanta, G. Morel, and
A. Beiras 1989b Localization of serotonin-like immunoreactivity
in the Merkel cells of pig snout skin. Anat. Rec. 225t267-271.
Gauweiler, B., E. Weihe, W. Hartschuh, and N. Yanaihara 1988 Presence and co-existence of chromogranin A and multiple neuropeptides in Merkel cells of mammalian oral mucosa. Neurosci. Lett.
Gottschaldt, K.-M., and C. Vahle-Hinz 1981 Merkel cell receptors:
Structure and transducer function. Science 214r183-186.
Gould, V.E., R. Moll, I. Moll, I. Lee, and W.W. Franke 1985 Biology of
disease. Neuroendocrine (Merkel) cells of the skin: Hyperplasias,
dysplasias and neoplasms. Lab. Invest. 52t334-353.
Gronblad, M., P. Liesi, and L. Rechardt 1983 Serotonin-like immunoreactivity in rat carotid body. Brain Res. 276:348-350.
Gu, J., J.M. Polak, F.J. Tapia, P.J. Marangos, andA.G.E. Pearse 1981
Neuron-specific enolase in the Merkel cells of mammalian skin.
The use of specific antibody as a simple and reliable histological
marker. Am. J . Pathol. 104t63-68.
Halaris, A.E., and D.X. Freedman 1977 Vesicular and juxtavesicular
serotonin: Effects of lysergic acid diethylamide and reserpine. J .
Pharmacol. Exp. Ther. 203575-586.
Hartschuh, W., and D. Grube 1979 The Merkel cell-a member of the
APUD cell system? Fluorescence and electron microscopic contribution to the neurotransmitter function of the Merkel cell granules. Arch. Dermatol. Res. 265:115-122.
Hartschuh, W., M. Reinecke, E. Weihe, and N. Yanaihara 1984 VIPimmunoreactivity in the skin of various mammals: Immunohistochemical, radioimmunological and experimental evidence for a
dual localization in cutaneous nerves and Merkel cells. Peptides
Hartschuh, W., and E. Weihe 1988 Multiple messenger candidates
and marker substances in the mammalian Merkel cell-axon complex: A light and electron microscopic immunohistochemical
study. Prog. Brain Res., 74t181-187.
Hartschuh, W., and E. Weihe 1989 Pancreastatin-like immunoreactivity in epidermal Merkel cells of pig and man. Neurosci. Lett.
98:258 -263.
Hartschuh, W., E. Weihe, M. Biichler, V. Helmstaedter, G.E. Feurle,
and W.G. Forssman 1979 Met-enkephalin-like immunoreactivity
in Merkel cells. Cell Tissue Res. 201r343-348.
Hartschuh, W., E. Weihe, and N. Yanaihara 1989 Immunohistochemical analysis of chromogranin A and multiple peptides in the
mammalian Merkel cell: Further evidence for its paraneuronal
function? Arch. Histol. Cytol. 52t423-431.
Hartschuh, W., E. Weihe, N. Yanaihara, and M. Reinecke 1983 Immunohistochemical localization of vasoactive intestinal polypeptide (VIP) in Merkel cells of various mammals: Evidence for a
neuromodulator function of the Merkel cell. J . Invest. Dermatol.
Horch, K.W., D. Whitehorn, and P.R. Burgess 1974 Impulse generation in type I cutaneous mechanoreceptors. J . Neurophysiol. 37:
Hsu, S.-M., L. Raine, and H. Fanger 1981 Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques. A comparison between ABC and unlabeled antibody (PAP) procedures.
J . Histochem. Cytochem. 29577-580.
Iggo, A,, and A.R. Muir 1969 The structure and function of a slowly
adapting touch corpuscle in hairy skin. J . Physiol. 200:763-796.
Ishida-Yamamoto, A,, E. Senba, and M. Tohyama 1989 Distribution
and fine structure of calcitonin gene-related peptide-like immunoreactive nerve fibers in the rat skin. Brain Res. 491t93-101.
Kanagawa, Y., T. Matsuyama, A. Wanaka, S. Yoneda, K. Kimura, T.
Kamada, H.W.M. Steinbusch, and M. Tohyama 1986 Coexistence
of enkephalin- and serotonin-like substances in single small intensely fluorescent cells of the guinea pig superior cervical ganglion. Brain Res. 379t377-379.
Katz, D.M., and I.B. Black 1986 Expression and regulation of cate-
cholaminergic traits in primary sensory neurons: relationship to
target innervation in vivo. J . Neurosci. 6:983-989.
Lane, E.B., and M. Whitear 1977 On the occurrence of Merkel cells in
the epidermis of Teleost fishes. Cell Tissue Res. 182:235-246.
Leon, J., and A.J. McComas 1984 Touch dome properties as a function
of age. Exp. Neurol. 84r274-282.
Lewis, J.C., and C.G. Moertel 1978 Platelet 5-hydroxytryptamine
storage in the carcinoid syndrome. An electron microscopic autoradiographic study. Am. J . Clin. Pathol. 70t628-631.
Merkel, F. 1875 Tastzellen und taskorperchen bei den hausthieren
und beim menschen. Arch. Mikr. Anat. 11:636-652.
Moll, R., I. Moll, and W.W. Franke 1984 Identification of Merkel cells
in human skin by specific cytokeratin antibodies: Changes of cell
density and distribution in fetal and adult plantar epidermis.
Differentiation 28t136-154.
Moll, I., R. Moll, and W.W. Franke 1986 Formation of epidermal and
dermal Merkel cells during human fetal skin development. J .
Invest. Dermatol. 87t779-787.
Ness, K.H., T.H. Morton, and B.A. Dale 1987 Identification of Merkel
cells in oral epithelium using antikeratin and antineuroendocrine monoclonal antibodies. J. Dental Res. 66t1154-1158.
Nurse, C.A., and J . Diamond 1984 A fluorescent microscopic study of
the development of rat touch domes and their Merkel cells. Neurosci. 11:509-520.
Nurse, C.A., K.M. Mearow, M. Holmes, B. Visheau, and J . Diamond
1983 Merkel cell distribution in the epidermis as determined by
quinacrine fluorescence. Cell Tissue Res. 228t.511-524.
Ortonne, J.P., and M. Darmon 1985 Merkel cells express desmosomal
proteins and cytokeratins. Acta Derm. Venereol. 65t161-164.
Ortonne, J.P., J.P. Petchot-Bacque, P. Verrando, A. Pisani, G.
Pautrat, and F. Bernerd 1988 Normal Merkel cells express a
synaptophysin-like immunoreactivity. Dermatologica 177tl-10.
Owman, C., R. Hlkanson, and F. Sundler 1973 Occurrence and function of amines in endocrine cells producing polypeptide hormones.
Fed. Proc. 32t1785-1791.
Pasche, F., Y. Merot, P. Carraux, and J.-H. Saurat 1990 Relationship
between Merkel cells and nerve endings during embryogenesis in
the mouse epidermis. J. Invest. Dermatol. 95t247-251.
Pearse, A.G.E. 1986 The diffuse neuroendocrine system: Peptides,
amines, placodes and the APUD theory. Prog. Brain Res., 68:
Pelto-Huikko, M., T. Salminen, and A. Hervonen 1985 Localization of
enkephalins in adrenaline cells and the nerves innervating
adrenaline cells in rat adrenal medulla. Histochemistry 82t377383.
Reeh, P.W. 1986 Sensory receptors in mammalian skin in an in vitro
preparation. Neurosci. Lett. 66t141-146.
Sanders-Bush, E., and L.L. Martin 1982 Storage and release of serotonin. In: Biology of Serotonergic Transmission. N.N. Osborne,
ed. John Wiley & Sons, New York, pp. 95-97.
Schultzberg, M., J.M. Lundberg, T. Hokfelt, L. Terenius, J . Brandt,
R.P. Elde and M. Goldstein 1978 Enkephalin-like immunoreactivity in gland cells and nerve terminals of the adrenal medulla.
Neuroscience 3:1169-1186.
Scopsi, L., B.-L. Wang, and L.-I. Larsson 1986 Nonspecific immunocytochemical reactions with certain neurohormonal peptides and
basic peptide sequences. J . Histochem. Cytochem. 34t1469-1475.
Scott, S.A., E. Cooper, and J . Diamond 1981 Merkel cells as targets of
the mechanosensory nerves in salamander skin. Proc. R. SOC.
Lond. 8211t455-470.
Simon, E.J., and J.M. Hiller 1989 Opioid peptides and opioid receptors. In: Basic Neuroschemistry. G. J . Siegel, ed. Raven Press,
New York, p. 271.
Smith, K.R. 1970 The ultrastructure of the human Haarscheibe and
Merkel cell. J . Invest. Dermatol. 54t150-159.
Smith, K.R., and B.J. Creech 1967 Effects of pharmacological agents
on the physiological responses of hair discs. Exp. Neurol. 19t477482.
Sternberger, L.A. 1986 The unlabeled antibody peroxidase-antiperoxidase (PAP) method. In: Immunocytochemistry. L.A. Sternberger, ed. John Wiley & Sons, New York, pp. 90-209.
Straile, W.E. 1960 Sensory hair follicles in mammalian skin: The
tylotrich follicle. Am. J . Anat. 106:133-147.
Wang, L., M. Hilliges, T. Jernberg, D. Weigleb-Edstrom, and 0. Johansson 1990 Protein gene produce 9.5-immunoreactive nerve
fibres and cells in human skin. Cell Tiss. Res. 261t25-33.
Warner, T.F.C.S., H. Uno, G.R. Hafez, J. Burgess, C. Bolles, R.V.
Lloyd, and M. Oka 1983 Merkel cells and Merkel cell tumors.
Ultrastructure, immunocytochemistry and review of the literature. Cancer 52t238-245.
Warren, R.P., K.K. Kane, R.A. Berger, and V.K. Singh 1990 Seroto-
nin-induced suppression of lymphocyte DNA synthesis and natural killer cell activity. Ann. N.Y. Acad. Sci. 594:429-431.
Winkelmann, R.K. 1977 The Merkel cell system and a comparison
between it and the neurosecretory or APUD cell system. J. Invest.
Dermatol. 69:41-46.
Yasargil, G.M., L. Macintyre, R. Doucette, B. Visheau, M. Holmes,
and J. Diamond 1988 Axonal domains within shared touch domes
in the rat: A comparison of their fate during conditions favoring
collateral sprouting and following axonal regeneration. J. Comp.
Neurol. 270r301-312.
Zaccone, G. 1986 Neuron-specific enolase and serotonin in the Merkel
cells of conger-eel (Conger-conger) epidermis. An immunohistochemical study. Histoehemistry 85:29-34.
Без категории
Размер файла
1 132 Кб
like, touch, merkel, serotonin, skin, neurons, dome, afferent, immunoreactivity, rats, hair, cells
Пожаловаться на содержимое документа