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Rat ovarian mast cellsDistribution and cyclic changes.

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THE ANATOMICAL RECORD 197: 489-493 (1980)
Rat Ovarian Mast Cells: Distribution and Cyclic
Laboratory of Compamtiue Reprcductwn, Department of Environmental, Population,
and Organismic Biology, University of Colomdo, Boulder, Colomdo 80309
Numerous tissue mast cells are present in the ovarian medulla
and hilus and in the oviduct of rats. In the medulla, most of these mast cells are
in the connective tissue of the stroma near blood and lymphatic vessels. During
proestrus, many of the medullary mast cells totally degranulate and thus are
not visible histochemically; they then regranulate during estrus. In contrast, the
number of stainable mast cells in the ovarian hilus and oviduct does not change
during the estrous cycle. Histofluorometric methods demonstrate that mast cells
in the ovarian medulla and hilus, as well as the oviduct, contain histamine. In
addition, the lining of small blood vessels in the ovarian medulla contains
histamine. Thus, mast cell and blood vessel histamine secretion may play a role
i n ovarian function.
In most mammals studied, ovarian blood plays a vital role in follicular growth and
flow markedly increases shortly before ovula- ovulation (Ellinwood e t al., 1978), may be
tion (see Ellinwood et al., 1978). More specif- mediated by release of ovarian histamine.
ically, ovarian blood flow in the rat is low That histamine plays a n essential role in
during diestrus and early proestrus, but in- ovarian follicular growth is supported by the
creases t o a peak at 2000 hr of proestrus. The inhibitory effects of exogenous antihistamines
flow rate remains high until early estrus, and on this phenomenon (Lipner, 1971; Knox,
then decreases to a low level at 1500 hr of 1974; Gergely et al., 19761, as well a s by the
estrus (Wurtman, 1964; Yoshinaga e t al., finding that cystic follicle formation is related
1969). Large, preovulatory mammalian folli- to hypersecretion of histamine (Hunter and
cles receive most of this increase in blood Leathem, 1968).
In mammals, histamine exists in bound
delivery (Ellinwood et al., 1978). The blood
vessels of rat preovulatory follicles become form in granules of tissue mast cells as well
hyperemic, and their permeability greatly in- as in circulating basophils and blood platelets
creases (Basset, 1943; Parr, 1974). In the rat, (see Beaven, 1978). Also, the endothelium of
as in other mammals (Ellinwood et al., 1978), small blood vessels synthesizes and secretes
the preovulatory surge of luteinizing hormone “inducible” histamine i n response to local,
(LH) appears to cause the proestrous increase intravascular conditions (Schayer, 1962). The
in ovarian blood flow (Ellis, 1961; Parlow and source of histamine within the ovary, howReichert, 1963; Szego and Gitin, 1964; Wurt- ever, is not known. In this paper, we report
man, 1964; Piacsek and Huth, 1971).
that the ovarian hilus and medulla of the
Histamine is a potent, vasoactive biogenic diestrous, estrous, and metestrous rat contain
amine, and the above-described effect of LH a large number of mast cells. In addition,
on ovarian blood flow in the rat is mimicked degranulation of medullary mast cells during
by histamine but not by follicle-stimulating proestrus results in a significant reduction in
hormone, prolactin, serotonin, epinephrine, or the number of these cells visible under the
norepinephrine (Wurtman, 1964). Exogenous light microscope. We also report that ovarian
LH causes depletion of ovarian histamine in mast cells contain histamine, as does the linthe rat (Szego and Gitin, 19641, and the anti- ing of small blood vessels present in the ovarhistamine promethazine hydrochloride blocks ian medulla.
the hyperemic effect of LH on rat ovaries
(Piacsek and Huth, 1971). Therefore, the LHReceived December 17. 1980. accepted March 5. 1980
induced increase in ovarian blood flow, which
@ 1980 ALAN R. LISS, INC.
We removed one ovary from each of 18
adult, cycling rats (200 gm, Sprague-Dawley).
Sample-size for each stage of the estrous cycle
(as determined by vaginal smears) was: proestrus, 6; estrus, 4; metestrus, 3; diestrus, 5.
Rats were killed in the late afternoon each
day of the estrous cycle. Each ovary was fixed
in 10% neutral-buffered formalin, embedded
in Paraplast, and serially sectioned a t 6 pm.
The sections were stained with toluidine blue,
which stains granules of mast cells (Humason,
1972). For each ovary, a total of 10-20 individual sections, each a t least 36-pm apart,
were scored to produce a mast cell index, or
MCI. A 0.083 mm2 square grid (400 X ) was
used. Each section was viewed and the grid
fixed over a n area of maximal mast cell concentration. All mast cells within the grid were
counted, and the average score from the 10-20
sections produced the MCI for an individual
ovary. An MCI for the ovarian medulla, the
ovarian hilus, and the oviduct was obtained
in this manner. We also scored each mast cell
as being either partially degranulated or
granulated. Our "partially degranulated"
stage was equal to stages 1 and 2 of Gibbons
and Chang (1972), and our "granulated" stage
was their stage 3. Finally, a n ovary from two
rats in each of the four stages of the estrous
cycle was examined for the presence of histamine using the histofluorometric method of
Enerback (1969).
x 8 O r
Fig. 1. Index of total mast cells (granulated plus partially degranulated) in the ovarian medulla, hilus, and
oviduct throughout the rat estrous cycle. Means S.E. D,
diestrus; P, proestrus; E, estrus; M, metestrus. For ovarian
medulla, analysis of variance detected significant variation
(F [3,141= 14.84,p < 0.001).Duncan's Multiple Range
Group Comparison Test indicated that E > D = M > P a t
p < 0.05. Numbers of mast cells in hilus and oviduct were
statistically similar across the estrous cycle.
Mast cells are present in medullary tissue
of the rat ovary. These cells also are abundant
in the connective tissue of the hilus and in the
TABLE 1. Distribution and condition
of mast cells in the rat ovarian medulla, hilus, and
ouiduct durinrr the estrous cycle.
.01 ? .01
.02 ? .02
.08 4 .04
.13 2 .05
.07 2 .04
.03? .03
.03 ? .02
.13 ? .04
.08 ? .03
.40 ? .13
.12? .04
Ovarian Medulla
.O1 2 .O1
.06? .03
.03 ? .01
"Means * S.E.
bConsidering all ovarian compartments, analyses of variance detected significant main-effects for granulation conditions IF (1.101 = 79.19, p < 0.001) and cycle stagp IF 13.14) = 14.48. p < 0.001) for ovarian
medulla There was also a significant interaction between cycle stage and granulation condition of mast cells
in the medulla (1.'[3.10]
= 290.49, p < 0.001). Duncan's Multiple Range Test detected significant differences
lp < 0.051 among degranulated cells in the ovary a t each stage of the estrous cycle.
49 1
interfascicular connective tissue of the oviduct. In contrast, no mast cells were seen in
the theca of ovarian follicles or corpora lutea.
The MCI for the ovarian medulla was significantly higher at estrus, moderate a t diestrus
and metestrus, and extremely low a t proestrus
(Fig. 1). In contrast, the MCI for the hilus and
oviduct was relatively high and similar
through the estrous cycle (Fig. 1).In all tissues
(ovarian medulla, ovarian hilus, and oviduct),
a great majority of the mast cells were partially degranulated a t all stages of the estrous
cycle (Table 1).
In the ovarian medulla, most of the mast
cells were in the connective tissue of the
medullary stroma, many were in the connective tissue surrounding venules, and a few
were adjacent to arterioles, lymphatic vessels,
and interstitial tissue (Table 1; Fig. 2). The
changes in the total MCI for the ovary during
the estrous cycle were reflected in each medullary compartment, and these changes were
mainly accounted for by differences in the
index for partially degranulated cells during
the cycle (Table 1).
Histofluorometric methods demonstrated
that histamine was present (yellow fluorescence) in the mast cells of the ovarian medulla
and hilus as well as the oviduct. Also, the
lining of arterioles and venules in the ovarian
medulla and hilus exhibited yellow fluorescence, indicating that these vessels contain
histamine. Our sample was not large enough
t o determine if the content of histamine in the
mast cells and vessels changed during the
estrous cycle.
.18 t .07
.01 t .01
.05 -c .02
.10 f .06
.78 f .23
.15 f .04
1.05 f .ll
.58 t .07
.06 t .03
+ .04
.ll t .07
It is clear that tissue mast cells are abundant in the ovarian medulla and hilus of the
rat. This is the first report of mast cells in the
rat ovary. In cow ovaries, a few mast cells
along with basophilic leucocytes and plasma
cells congregate in the thecal interstitial
spaces of preovulatory follicles (Cupps et al.,
1959). A similar event occurs in the rabbit
ovary (Zachariae et al., 1958; Bjersing and
Cajander, 1974).Our observation that no mast
cells occur around growing or preovulatory
rat follicles, the finding of Peppler and Greenwald (1970) that ovaries of diestrous rats contain no preovulatory follicles, and the report
of Szego and Gitin (1964) that diestrous rat
ovaries contain high amounts of histamine,
all suggest that mast cells in the ovarian
medulla or hilus and not in preovulatory follicles are a source of ovarian histamine.
The blood vessels in the medulla and hilus
also contain “inducible” histamine, and the
relative roles of mast cell histamine and inducible histamine in influencing ovarian blood
flow are not known. Nevertheless, the presence of variation in mast cell number during
the estrous cycle suggests that the mast cells
degranulate during proestrus and are involved
in the increase in ovarian blood flow during
this stage of the cycle. Mast cell degranulation
is a normal stage in the secretion of histamine
(Goth and Johnson, 1975). We detected a decrease in mast cell number during proestrus,
and an increase during estrus. These changes
in number could reflect changes in synthesis
and release of granules, with cells completely
.26 + .09
.03 t .03
.03f .03
.05 .05
5.38 t -99
5.85 f .46
6.55 f .77
5.29 + .65
.77 t .26
.03 t .03
.12 t .12
.63 f .16
5.22 f .64
6.40 t .28
5.88 + .64
5.75 + .61
‘Considering a l l mast cells. analyses of variance detecLed significanL main effects far ovarian compartment [F14.70) = 27.15.
p < 0.0011 and stageof theestrouscyclelF[3.14] = 14 R4.p < 0 OOl).Theinteraction hetweenstageotcycleandovariancompartment was not significant. Among the ovarian compartment, Duncan’s Multiple Range Test detected t h a t the mast cells in
stromal Connective tissue > venules = Interstitial = arterioles = lymphatics ( p < 0.05).
Fig. 2. Mast cells (-+) in the ovarian medulla of an estrous rat. Note the arteriole (A) and interstitial tissue (IT).
Toluidine blue, x 400.
discharging granules being invisible using toluidine dye. Even though this dye stains heparin (and not histamine) in the granules
(Beaven, 1978), histamine is released with
heparin as granules are discharged.
The LH surge during the late afternoon or
evening of proestrus in the rat (Daane and
Parlow, 1971; Butcher et al., 1974) may cause
the lowering of mast cell number during
proestrus, because this hormone may stimulate mast cell degranulation. The mast cells
then regranulate during estrus. Whatever factor causes ovarian mast cell degranulation, its
effects may be limited to the medulla, since
mast cells in the ovarian hilus and oviduct do
not change in number during the estrous
cycle. Whether LH influences the secretion of
inducible histamine in ovarian blood vessels
is not known. However, both compound 48/80
(which degranulates mast cells) and an antihistamine are needed to inhibit ovarian follicular growth in the rat (Lipner, 1971), suggesting that inducible histamine as well as
mast cell histamine play a role in ovarian
function in this species.
Our discovery that the rat ovary contains
mast cells that vary in number during the
estrous cycle poses more questions. For example, what are the changes in mast cell
condition throughout the day of proestrus?
Does exogenous LH induce ovarian mast cell
degranulation followed by regranulation?
What are the roles of mast cell and inducible
histamine in ovarian function? Further research should be conducted to answer these
We thank David 0. Norris and Kevin Fitzgerald for criticizing this manuscript. Supported by a research grant (HD 11482-02) and
a Research Career Development Award (HD701650 5 ) from the National Institute of Child
Health and Human Development.
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