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© 1985S. K argcr A G . Basel
(HKM—5180/85/1213-0153 S 2.75/0
Acta anat. 121: 153-162 (1985)
Target Cells of Vitamin D in the Vertebrate Retina
D. S. Schreiner^'. S. S. Jamie", D. E. M. Lawsonb
*Department of Anatomy. University of Ottawa. Ont., Canada; ’’University of Cambridge and Medical Research Council. Cambridge. UK
Key Words. Vitamin D •Calcium-binding protein • Vertebrate retina
The classical targets for vitamin D were once thought
to comprise only organs that are directly involved in
calcium homeostasis, e.g. bone, intestine, kidney, para­
thyroids, laying hen shell glands, etc. Recently, some
new vitamin D target organs have been uncovered by a
variety of modern techniques such as: (a) the biochemi­
cal isolation and characterization of high affinity recep­
tors for 1.25 (011)2 D3 [Brumbough and Haussler, 1974;
Lawson and Wilson, 1974; Christakos and Norman,
1980a, b; Pike et al., 1980] which is the most potent
metabolite of vitamin D; (b) the autoradiographic
localization of labelled 1.25 (OH)2D3[Stumpf et al., 1979,
1980; Narbaitz et al.. 1981], and finally (c) the detection
of a specific protein synthesized as a result of genomic
stimulation by vitamin D,. Vitamin D-dependent cal­
cium-binding protein (D-CaBP) is the only well known
protein whose synthesis is de novo under the influence
of 1,25 (OI I)2 D3 [Wassennan and Taylor, 1966; Enilage
et al., 1973; Spencer et al., 1976; Christakos and Nor­
man, 1980a. b]. Although the amount of D-CaPB in
various tissues can be measured by radioimmunoassay
[Christakos et al., 1979; I'homasset et al., 1982], the
localization of D-CaBP in specific cell types can be car­
ried out by immunocytochemical methods [Morrissey et
al., 1978; Jande et al.. 1981a, b; Schreiner et al.. 1983].
Results obtained with these three different techniques
are in good agreement as to the identity of vitamin D
target tissues and cells.
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Abstract. Using PAP technique, cellular localization of vitamin D-dependent calcium-binding protein (D-CaBP)
was investigated in vertebrate retina with monospecific antisera against chick duodenal D-CaBP. In the chick
retina, the receptor cells were positive. In the inner nuclear layer, horizontal cells and some bipolar cells were also
positive. Some amacrine cells as well as different levels of the inner plexiform layer were also positive for D-CaBP.
A few interspersed ganglion cells were positive but their axons forming the optic tract were negative. Muller's cells
were negative. In 1-day-old chicks and 4-week-old rachitic chicks there was paucity and absence, respectively, of DCaBP staining in horizontal cells. In the mouse, rat, and rabbit the receptors had only trace amounts of reaction
product in their outer segment and pedicle. Horizontal cells were densely positive throughout their cellular body
and processes. Some amacrine cells in the inner nuclear layer were positive. In the mouse and rat three horizontal
levels of the outer plexiform layer were very prominent because of their dense staining for D-CaBP. Many ganglion
cells were also positive along with their axons forming the optic nerve. In the rabbit, no positive layers were seen in
the inner plexiform layer, and ganglion cells with their fibers were negative. In the frog retina there were smaller
amounts of D-CaBP in the receptor cells and horizontal cells than that of the chick retina. Also, the fibers of the
ganglionic cells were positive for D-CaBP. In all species studied, some amacrine cells were stained for D-CaBP.
Because of its possible roles in membrane calcium transport and intracellular Ca++ regulation, it has perhaps similar
functions in these positive cells. The synthesis of D-CaBP is dependent upon vitamin D. These positive cells are thus
target cells of vitamin D.
Fig.l. Vertical section of a chick
retina incubated with non-immune
rabbit serum (control). All the com­
ponents of the retina including re­
ceptor cells (RC). the outer nuclear
layer (ONL). the outer plexiform
layer, the inner nuclear layer (INL).
the inner plexiform layer (IPL). the
ganglion cell layer (GC). and the
fiber layer (FL) were negative.
Material and Methods
4-week-old while Leghorn chicks on normal diel and some on
rachitogenic diet [Barnes el at., 19731; I-month-old Sprague-Dawley
rats; black mice (C57BL/6.I Jackson Labs); Paris R3 white mice; 2month-old New Zealand white rabbits, and frogs <liana pipiens) were
used as source for retina. These were decapitated and each eye was
quickly removed and transferred into fixative. The eyes were
hemisected at the equator, the anterior half and the vitreous humor
was removed. Fixation was carried out in Carnoy's fixative for 3 h at
room temperature |Jande et al.. 1981a]. The retinas were then
thoroughly washed in 90% ethyl alcohol and processed for paraffin
sectioning. 5-um thick sections were used for histochemical staining.
The PAP method for paraffin sections according to Sleinberger
11979] was followed. Sections were pre-adsorbed with a 3% solution
of normal goat serum (Gibco). The primary antiserum (anti-D-CaBP)
was diluted 'Am, the link antiserum (anti-rabbit goat IgG. Miles
Laboratories) was diluted Vx and the PAP complex (rabbit peroxidase
anti-peroxidase. Miles Laboratories) 'Aw. All dilutions were done with
0.05 M Tris-HCI saline containing 1% normal goat scrum. The slides
were incubated in 0.5% diaminobenzidine-HCI containing 0.01% U4),
for 20 min at room temperature. Endogenous peroxidase was inhi­
bited according to the method of Heydermtin and Neville 11977], Con­
trol sections were stained with non-immune rabbit serum diluted 'A...
instead of the antisera. All sera were decomplemented for 30 mm at
56 °C.
Monospecific antisera against chick duodenal D-CaBP was
prepared according to Spencer et al. 11976] in the laboratory of
Dr. !). E. M. l.awson.
The localization of D-CaBP was carried out in the
chick, rat. mice, rabbit and frog. The best results were
obtained for the chick retina and these will be presented
Normal Chicks
Control sections, incubated with normal rabbit
serum, did not show any reaction product in the entire
retina (fig. 1). The histology of the chick retina is well
known [Hodges, 1974] and the positive retinal neurons
have been identified according to their location. Inner
to the pigment epithelium is the receptor cell layer
which has been shown to contain various morphological­
ly distinct types of photoreceptor cells [Meyer and
Cooper, 1966; Morris and Shorey, 1967], All receptor
cells have been found to contain the dark reaction prod­
uct indicative of the presence of D-CaBP. Various struc­
tural regions of the receptors namely, the inner seg­
ment. the cytoplasm surrounding the nucleus, some of
the nuclei and all synaptic pedicles have been found to
be very positive for D-CaBP (fig. 2, 3). There was a
sharp demarcation at the outer limiting membrane, be-
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D-CaBP which was originally isolated from chick
duodenum [ Wasserman and Taylor, 1966] has been very
well characterized [Ingersoll and Wasserman, 1971;
Bredderman and Wasserman, 1974; Fullmer and Was­
serman, 1975] and has also been demonstrated in the
brain [Taylor, 1974, 1976]. In the central nervous system
D-CaBP is confined to neurons and their processes of
certain specific nuclei [Jamie et al., 1981a, b; Roth et al.,
The retina, which is an extension of the diencepha­
lon. has been found to also be a target organ of vitamin
D and this communication describes the cellular dis­
tribution of D-CaBP in various retinal cell types and
endeavours to discuss the functional significance of
these new target cells its it relates to calcium
Vitamin D. Calcium-Binding Protein and Vertebrate Retina
Fig.2. Vertical section of a nor­
mal chick retina stained with anti-DCaBP. The inner segment of the re­
ceptor cells (RC) are positive for DCaBP. At the outer limiting mem­
brane (OLM) there is a sharp bound­
ary because of the lighter staining of
outer nuclear layer (ONL). In the
outer plexiform layer (OPL) the
positive pedicles of the RC and the
positive processes and cellular
bodies of horizontal cells (HC) from
two distinct layers. In the inner half
of the inner nuclear layer (INI.) posi­
tive amacrine cells (AC) are seen. In
the inner plexiform layer (IPL) at
least five horizontal bands can be
distinguished. A few scattered gan­
glion cells (GC) are positive. The
fiber layer (FL) and the inner limit­
ing membrane (INC) are negative.
cause the outer nuclear region showed smaller amounts
of D-CaBP (fig.3). The amount of cytoplasm in this
layer is sparse since most of the space is taken up by the
nuclei, of which only a few were positive and thus it
stained lightly. The synaptic pedicles always stained
dark and appear as a distinct layer. Müller cells, which
take part in the formation of the external limiting mem­
brane through their junctional complexes with the re­
ceptor cells, did not show any staining for D-CaBP
The outer plexiform layer where the pedicles of the
receptor cells form synaptic connections with the hori­
zontal and bipolar cells stained quite dark (fig.3). Quite
often in the outer plexiform layer two stratifications
were distinguishable: an outer one which represents the
synaptic pedicles of the photoreceptors and an inner one
which is composed of the cell processes of horizontal
and bipolar cells. Further inwards is the inner nuclear
layer (INL) which contains four types of cells; horizon­
tal cells, bipolar cells. Müller cells and amacrine cells.
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Fig.3. Higher magnification of
normal chick retina stained with
anti-D-CaBP. Here again the inner
segments of the receptor cells RC
are clearly positive. The outer
nuclear layer (ONL) stains lightly.
I'he pedicles of the RC arc positive.
In the inner nuclear layer (1NL).
horizontal cells (HC) and probably
some bipolar cells (BC) are also
positive. Amacrine cells (AC) con­
taining D-CaBP arc present close to
the INL. One AC in particular is
labeled and its sole process is seen
reaching into a D-CaBP positive lay­
er of the inner plexiform layer,
x 562.5
The main cell body of the horizontal cell, including the
nucleus and the cellular processes taking part in the in­
ner plexiform layer were positive for D-CaBP (fig. 2, 3).
It appears that the major amount of reaction product
was on the outer side of each nucleus (fig. 3). The nuclei
of the bipolar cells are situated in the outer half of the
INL. Only a few nuclei in these regions were positive
(fig.3). Muller's cells which begin at the outer limiting
membrane and end at the inner limiting membrane were
completely negative. Further inwards in the INL. closest
to the inner plexiform layer (IPL), are the amacrine cell
bodies. Although it is not possible to identify the various
types of these cells as described by Cajal [1972], it is very
apparent that some of the D-CaBP-positive cells are
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Fig. 4. Section of a retina from a
4-week-old rachitic chick, stained
with anti-D-CaBP. In the outer plcxiform layer (OPL) only the pedicles
of the receptor cells are clearly posi­
tive and the horizontal cells are
negative. In the inner nuclear layer
(INL). scattered positive cells arc
present: some can be clearly iden­
tified as amacrinc cells, others are
probably bipolar cells. Note some
positive ganglion cells. x225.
Fig.5. Tangential section of a re­
tina from a 4-week-old rachitic chick
stained with anti-D-CaBP. The inner
segment of receptor cells (RC) as
well as the cytoplasm in the outer
nuclear layer (ONI.) are positive.
Horizontal cells in the outer plexiform layer (OPL) are not clearly
seen and probably some bipolar cells
(some of the group are labeled as
BC) are positive. x225.
Fig. 6. Section of a retina from a
1-day-old chick stained with anti-DCaBP. The receptor cells, probably
some bipolar cells (BC). and some
amacrinc cells (AC) and ganglion
cells are positive. The horizontal
cells (HC) are only faintly positive.
F’ig.7. Tangential section of a re­
tina from a I-day-old chick stained
with anti-D-CaBP. In the outer
nuclear layer (ONL) the cytoplasm
and the nuclei of the receptor cells
are positive. In the inner nuclear lay­
er (INL). horizontal cells (HC) are
faintly positive while probably some
bipolar cells (BC) and some amacrine cells at the bottom of the INL
arc positive. X365.
Vitamin D. Calcium-Binding Protein and Vertebrate Retina
Rachitic Chicks
The overall staining for D-CaBP in retina of chicks
on rachitogenic diet for 4 weeks w'as lower than that in
retina of chicks on normal diet. The receptor cells
showed less reaction product but each cell's synapticpedicle was positive and thus the outer region of the
outer plexiform layer, as mentioned earlier for the nor­
mal retina, were darkly stained (fig.4). The horizontal
cells, their main cell body and the nucleus and their
cellular processes in the outer plexiform layer, were
completely negative (fig.4). This is very clear in the
tangential section of the retina (fig.5). Thus the inner
stratification of the OPL was not visible with D-CaBP
staining. Some of the bipolar cells and some amacrine
cells were positive (fig.4. 5). The various layers in the
IPL were less distinct (fig.4). Some ganglion cells were
positive. No other major differences were found from
the normal retina.
An examination of the retinal development was car­
ried out on I-day-old chicks and on 2-week-old chicks
on normal diet and 2-week-old chicks on rachitogenic
diet. At day 1, different parts of the receptor cells, espe­
cially their regions situated in the outer nuclear region,
w'ere positive for D-CaBP (fig.6). The horizontal cell
bodies along with their nucleus and their extensions to
the OPL were faintly positive (fig.6. 7). Some amacrine
cells and probably some bipolar cells were also distinctly
positive (fig.6, 7). From the INL inwards, no difference
was observed from the normal 4-week-old chick retina.
In the retina of the 2-week-old chicks on a normal diet,
only some receptor cell bodies in the ONL region were
seen to be positive (fig. 8). The two stratifications of the
OPL. the outer one formed by receptor cells synaptic
pedicles and the innermost layer formed by horizontal
cell processes, were visible since the horizontal cells
were also positive (fig. 8). No major distinctions w'ere
observed in the inner layers of the retina when com­
pared to 4-week-old chick retina. In the retina from
chicks fed a rachitogenic diet for 2 w'eeks the cellular
bodies and the cytoplasm in the synaptic pedicle of the
receptor cells were positive (fig.9). The soma and the
processes of the horizontal cells that take part in the
formation of the outer plexiform layer were completely
negative. Thus of the two components in the OPL, the
outer one formed by synaptic pedicles was distinct while
the inner one composed of the horizontal cell processes
was not visible (fig.9). Further inwards, amacrine cells
and probably some bipolar cells were positive (fig. 9).
The horizontal lines in the IPL were present (fig. 9). No
major differences we re noted in the inner regions of the
normal and rachitic retina.
Mouse and Rat
In the retina of the white and the black mouse and
also in that of the rat. similar neuronal types were posi­
tive. Thus the following description applies to these
three rodents. Only barely detectable amounts of DCaBP were present in the receptor cells -(fig. 10. 11).
The horizontal cells soma and their processes stained
very darkly, making the OPL very prominent (fig. 10.
11). In the INL. the cell body and nucleus of many
amacrine cells were positive (fig. 10, II). In some of
these cells, the single main process was seen to reach
into the IPL (fig. 10. 11). Three horizontal lines in the
IPL were very prominent because of their dense staining
for D-CaBP. All the ganglion cells along with their
axons, which form the optic tract, were positive (fig. 10.
11 ).
In the rabbit retina (fig. 12. 13) as in the mouse and
rat, only a very faint staining was observed in the inner
segment and none was visible in the receptor cell pedi­
cles. Here again the OPL was the most prominent layer
mainly because of the dark positive staining in the hori­
zontal cells. The INL is very thin and bipolar cells along
with Müller cells were negative. However, some amac-
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indeed amacrine cells (fig.3). Their location in the INL
and their mode of branching in the IPL support such a
conclusion. In a number of positive amacrine cells their
cellular process can be seen extending into the IPL
where they branch and take part in synaptic connections
(fig. 3).
In the avian retina, the IPL is made up of many hori­
zontal layers as a consequence of the stratification of the
synapses between the processes of bipolar, amacrine
and ganglion cells. I lowever. five layers, two being very
prominent, were visible because of the positive staining
for D-CaBP (fig. 2. 3).
Some of the ganglion cells were found to be positive.
The reaction product was seen in some nuclei as well as
in the cytoplasm (fig.2). The distribution of positive
cells varied a great deal and thus in sonic parts of a
section only a few while in other regions many positive
cells were observed. The cellular processes of the gan­
glion cells that take part in the IPL and others that form
the fiber layer, were negative (fig.2). The inner limiting
membrane formed by the processes of Muller's cells was
negative (fig. 2).
fine cells were positive (fig. 12). No horizontal stratifica­
tion stood out in the IPL as described in the chick and
other rodents (fig. 12).
The outer segment of the frog receptor cells as well as
their pedicles were faintly positive (fig. 14). Some dis­
tinct horizontal cells stained darkly. In the INL the cyto­
plasm as well as nuclei of many bipolar cells were posi­
tive (fig. 14). Some amacrine cells at the border of the
INL and IPL were quite positive (fig. 14). In the IPL
many sublayers stood out because of their positive stain­
ing (fig. 14). Many ganglion cells along with their axons
forming the optic tract were positive (fig. 14).
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Fig. 8. Section of a 2-week-oid
normal chick retina stained with
anti-D-CaBP. In the outer nuclear
layer (ONL) some of the cell bodies
of receptor cells are positive. In the
inner nuclear layer (INL) horizontal
cells, probably some bipolar cells
and some amacrine cells arc positive.
In the inner plexiform layer (IPL)
three positive bands are visible.
Fig. 9. Section of a 2-week-old
rachitic chick retina stained with
anti-D-CaBP. Receptor cell outer
segments and cell bodies in the outer
nuclear layer (ONL) are positive.
RC pedicles also stained for D-CaBP
but horizontal cells (HC) arc nega­
tive. Probably some bipolar cells
(BC). some amacrine cells and some
ganglion cells are positive. x365.
F'ig. 10. Sections of mouse retina
stained with anti-D-CaBP. The re­
ceptor cells (RC) have but a trace
amount of D-CaBP. In the inner
nuclear layer (INL) horizontal cell
bodies and the processes are densely
stained. Some amacrine cells (AC)
arc also positive. In the inner plex­
iform layer three horizontal lines are
distinctly stained. Most ganglion
cells (GC) and their fibers (FL) arc
positive. x225.
F'ig. 11. Section of a rat retina
stained with anti-D-CaBP. Here
again receptor cells are very lightly
stained. Horizontal cells (HC) and
some amacrine cells (AC) arc posi­
tive. Most ganglion cells (GC) and
their fibers (FL) are positive. x225.
Vitamin D, Calcium-Binding Protein and Vertebrate Retina
The presence of D-CaBP in the central nervous sys­
tem has been well documented [Taylor, 1974: Jmule et
ah, 1982; Roth et ah, 1981; Baimbridge et ah, 1980,
1982; Feldman and Christakos, 1983]. Since the retina is
an extension of the diencephalon, it is not surprising
that some retinal neurons contain D-CaBP. Here is thus
a newly discovered target organ for vitamin D in the
species investigated.
Earlier studies with epithelial tissues such as intes­
tine, hen shell gland and kidney have connected the
function of D-CaBP with transcellular calcium transport
as seen in calcium absorption and egg shell formation
[Jande et ah, 1981a, b. 1982]. This view is clearly not
supported by the presence of D-CaBP in so many cell
types such as epithelial reticular cells of the thymus, the
macrophage-like cells in spleen, the neuronal cells in the
brain, follicular cells of the ovary [Jande et ah, 1982]
which apparently are uninvolved in such processes. It
seems that D-CaBP is involved in more than one phy­
siological function in which case it may have a single
molecular action which has been incorporated into a
variety of processes. The latter almost certainly involves
the membrane transport of calcium but whether DCaBP is involved at this point or in the subsequent need
to regulate intracellular Ca2+ levels still remains unclear.
The presence of D-CaBP in various neurons in the reti­
na suggests special needs of these cells for calcium which
arc dependent on this protein either for membrane
transport and/or for intracellular Ca’+ concentration.
In all the species observed there was a wide variation
in the concentration of D-CaBP in the receptor cells.
Staining was deepest in the chick receptor cells and
weakest in those of the albino rat. There is evidence that
calcium acts as a transmitter in light induction of recep­
tors [Hagina, 1972; Yoshikami et al.. 1980; Hendriks et
al., 1974], The wide difference in D-CaBP observed in
chick and rat retina may represent different Ca:+ needs
which can only be solved by D-CaBP via its functions
outlined above.
Our developmental studies on normal and rachitic
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Fig. 12. Section of a rabbit retina
stained with anti-D-CaBP. Receptor
cells (RC) are very faintly positive.
Horizontal cells are the most densely
stained cells. Some amacrine cells
are also positive. x225.
Fig. 13. Partially tangential sec­
tion of a rabbit retina showing the
extensive branching of D-CaBP posi­
tive horizontal cell processes. x225.
Fig. 14. Section of a frog retina
stained with anti-D-CaBP. The re­
ceptor cells (RC) outer segments, as
well as their cellular bodies in the
outer nuclear layer (ONL) and their
pedicles are faintly positive. Hori­
zontal cells (HC). bipolar cells and
some amacrine cells (AC) in the in­
ner nuclear layer (INL) are positive.
Four horizontal lines in the inner
plexiform layer are positive. Many
ganglion cells (GC) and their fibers
(FL) are positive. x225.
Schreiner/J ande/Lawson
ous structures still remains to be solved. Thus in the
vertebrate retina various neurons have been demon­
strated to be the target cells of vitamin D through the
localization of D-CaBP in these ceils. The proper func­
tion of this protein in these cells still remains unclear.
Thanks are due to MRC of Canada (Grant No. MT 3089) and
MRC of UK for support of the research. Ms. Schreiner is holder of an
MRC Studentship.
Araki. M.; M aeda.T.; Kimura, H.: Dopaminergic cell differentiation
in the developing chick retina Rrain Res. 10: 97-103 (1983).
Arnold. B.M.; Kuttner, M.; Willis. D .M .; Hitchman. A .J.W .: Har­
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1135-1140 (1975).
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Baimbridgc, K.G.; Selke, P.A.: Ferguson, M.; Parkcs, C.O.: Hu­
man calcium-binding protein; in Seigel, Carafoli, Kretsingcr. MacLennan. Wasserman. Calcium-binding proteins; structure and
function, pp. 401-404 (Elsevier/North Holland. Amsterdam 1980).
Barnes. M .J.: Constable. B.J.; Morten, L .F.; Kodicek. E.: The in­
fluence of dietary calcium deficiency and parathyroidectomy on
bone collagen structure. Biochim. biophys. Acta 328: 373-382
Buckerfield. M.; Olivier, J.; Chubb, I.W.; Somatostatin-like iinmunoreactivity in amacrine cells of the chicken retina. Neurosci­
ence 6: 689-695 (1981).
Brecha. N.; Karten. H .J.; Laverack. C.: Enkephalin-containing
amacrine cells in the avian retina: immunohistochcmical localiza­
tion. Proc. natn. Acad. Sci. USA 76: 3010-3014 (1979).
Bredderman. P. B. ; Wasserman. R. H.: Chemical composition, affini­
ty for calcium and some related properties of the vitamin D-dependent calcium-binding protein. Biochemistry 13: 1687-1694 (1974).
Brumbough. P .F .; Haussler, M R.: la,25-Dihydroxycholecalciferol
receptors in intestine. 1. Association of la.25-Dihydroxycholecalciferol with intestinal mucosa chromatin. J. biol. Chem. 249:
1251-1257 (1974).
Cajal, S.R.Y .: The structure of the retina (Thomas, Springfield
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a radioimmunoassay for vitamin D-dependcnt chick intestinal cal­
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retina of chicks clearly demonstrate the dependency for
the synthesis of D-CaBP in the horizontal cells on vita­
min D. The horizontal cells take part in the formation of
OPL and the complexity of this layer differs in certain
species. These differences are related to the analytical
capacity of the retina [Michael, 1969], It appears that
horizontal cells in their function have special Ca:+ needs
that are dependent on D-CaBP.
In all the vertebrates observed, some amacrine cells
in the retina were always positive. The vertebrate retina
has been shown to consist of several types of amacrine
cells containing various different peptides such as:
acetylcholine [Nichols and Roelle, 1968), glycine,
GABA [Marshall and Voaden, 1974a, b] dopamine
[Araki et al., 1983], indolamine [Hauschild and Lalies,
1973; Florén, 1979). enkephalin [Brecha ct ah. 1979;
Tornquist et ah, 1981], somatostatin [Kirsch and
Leon hardi, 1979; Buckerfield et ah, 1981 ; Eskay et ah,
1980; Tornquist et ah, 1981; Ellis et ah, 1983], TRH
[Eskay et ah, 1980; Tornquist et ah, 1981], substance P
[Fukuda et ah, 1981 ; Karlen and Brecha, 1980; Eskay et
ah, 1980] glucagon and neurotensin [Tornquist et ah,
1981]. Evidence concerning the role of these neuropep­
tides also found in other areas of the brain has suggested
that these may function as neurotransmitters [Tornquist
et ah. 1981], In the retina, the release of TRH, somatos­
tatin and substance P under depolarizing conditions is
calcium dependent [Eskay et ah, 1980], and this could
be related to the functions of D-CaBP. Experiments are
under way in order to find out whether these D-CaBP
containing amacrine cells represent those that also con­
tain these neuropeptides.
In all the speeies observed except for the rabbit,
some horizontal layers in the IPL were densely positive.
These represent the synaptic contacts between D-CaBP
positive amacrine, bipolar and ganglion cells. Thus, like
the OPL, the IPL formed by the synaptic complexes of
bipolar, amacrine and ganglion cells, have special cal­
cium needs which as stated earlier, may require DCaBP.
In the rat, mouse and frog, most of the ganglion cells
along with their dendritic fibers were positive for DCaBP. However, in the chick only a few ganglion cells
were positive and their fibers were completely negative.
This correlates well with the observations of Feldman
and Christakos [1983] who noted that the rat optic tract
fibers were positive for D-CaBP and of Roth ct al.
[1981] who demonstrated that chick optic tract fibers
were negative. The functional significance for this dis­
parity in D-CaBP content between these two homolog­
Christakos. S.; Norman, A. W.: Vitamin D-dependenl calcium bind­
ing protein and its relation to 1,25-Dihydroxyvitamin D receptor
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Received: February 25. 1984
Accepted: April I, 1984
D. S.Schreiner, MD.
Department of Anatomy.
University of Ottawa,
Ottawa, Om. K1H 8M5 (Canada)
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