I I. pp.

Journal of Srrraid Biochemisrr): Vol. 781 to 790 Per&w~~on Press Ltd 1979. Printed in Great Britain

INTRAOVARIAN BIOSYNTHESIS

CONTROL OF PROGESTERONE BY GRANULOSA CELLS AND CORPUS LUTEUM

G. L. KUMARI* and C. P. CHANNING Department of Physiology, University of Maryland School of Medicine, 660 West Redwood Street, Baltimore, MD 21201, U.S.A. SUMMARY

Some of the recent developments on the intraovarian control of follicular development and luteinization have been discussed. Luteinizing hormone (LH) and follicle-stimulating hormone (FSH) regulate the growth and maturation of the follicle. However, the locus of control, by which the responsiveness of follicles to LH and FSH is regulated at different stages of follicular development, resides within the ovary itself. Granulosa cells obtained from small follicles have more FSH receptors and can accumulate more cyclic AMP and progesterone in response to FSH compared to cells obtained from large mature follicles. The situation is reversed in the case of LH responsiveness; larger follicles are more responsive than small ones. The steroidogenic potential of follicular cell types to secrete oestrogen and progesterone is altered with the stage of the follicle. In oiuo studies in the rhesus monkey demonstrated that thecal cells of the preovulatory follicle are the principal source of oestrogen and progesterone of the ovarian venous blood. In in vitro cultures, thecal cells alone could secrete more oestrogen than granulosa cells, and mixing of the two cell types augmented oestrogen synthesis. Addition of exogenous testosterone (0.15 pgg/ml) to granulosa cell cultures resulted in increased secretion of oestrogens. Granulosa cells in culture produced more progesterone than thecal cells. These findings suggest that the thecal layer is the main source of ovarian vein levels of oestrogen and progesterone and the two cell types may be responsible for the high levels of these steroids in the follicular fluid. A luteinization inhibitor, present in the follicular fluid of small and medium-sized follicles may regulate the responsiveness of follicles to gonadotrophins and prevent luteinization of granulosa cells prior to ovulation. Evidence for the presence of another inhibitor, a LH receptor binding inhibitor (LHRBI) in frozen aqueous extracts of porcine corpus luteum and its ability to inhibit progesterone secretion by cultured granulosa cells of mature porcine follicles has also been presented. MATURATION

OF THE FOLLICLE

The importance of pituitary LH and FSH in regulating ovarian processes such as maturation and rupture of the follicle and its luteinization have been reviewed elsewhere [l-4]. However, the mechanism by which a primary follicle is chosen to mature and ovulate under the influence of gonadotrophins is still a mystery. Since all the follicles are exposed to the same circulating levels of FSH and LH, the mechanism by which a follicle is selected must reside within the ovary itself. Intraovarian regulation of follicular maturation and luteinization will be discussed with special emphasis on the role of local steroidal and non-steroidal factors. (i) Oestrogen and progesterone synthesis in follicular cells Follicular development and maturation are under the influence of gonadotrophins and the mature follicle secretes oestrogen and progesterone after exposure to FSH and LH. Since the follicle is composed of two cell types, the inner granulosa cell layer and an outer thecal layer, the steroidogenic potential of both cells individually and in combination needs to be * Present Address: Department of Reproductive Biomedicine National Institute of Health & Family Welfare D-5 Green Park, New Delhi-110016, India.

assessed before other factors which control follicular maturation can be examined (Fig. 1). In the monkey [%7], as well as in the human [g-11], both cell types have been cultured to study their ability to secrete oestrogen and progesterone at various maturational stages. It has been shown that granulosa cells of small immature follicles (l-2 mm) have no ability to luteinize, but those of medium sized (3-5 mm) follicles will do so, in the presence of LH or FSH or both. Granulosa cells harvested from mature preovulatory (Graafian) follicles, however, luteinized spontaneously in culture and secreted elevated amounts of progesterone [5-7,12-151. The granulosa cells enclosed in the preovulatory follicles are thus prevented from luteinizing by some compound(s) present in the follicular fluid, which will be discussed later. It has been well documented that the preovulatory follicle is the principal source of preovulatory oestrogen in normal cycling mammals [16-201. Conflicting evidence is available as to which cell types contribute towards oestrogen synthesis and its secretion. into follicular fluid and the circulation. Falck[21] proposed, based on the observations of vaginal cornification index as an indication of oestrogen synthesis after transplanting both cell types in the anterior chamber of the eye of the rat, that both the thecal and granulosa cells complement one another in oestrogen synthesis. In vitro studies [22,23] support the

781

G. L. KUMARI and

782

SMALL FOLLICLE

C. P.

CHANNING

LAME PREOVULATORY FOLLICLE

Fig. 1. Diagrammatic sketch of the actions of FSH and LH on follicular development and maturation. Pg E2 = prostaglandin E2. Data taken from Channing and Tsafriri[l] with permission. “two-cell theory” of ovarian oestrogen secretion which indicates that the thecal cells secrete androgens which are transported to granulosa cells and converted to oestrogens. However, the electron microscopic observations [24,25] on the cell types of the preovulatory human and monkey follicle do not support an active role of granulosa cells in follicular steroidogenesis. These cells had little smooth endoplasmic reticulum and poorly developed mitochondrial cristae. It is difficult to logically explain the two cell theory under in oh conditions where there is probable transport of androgens from a well vascularized thecal compartment through a basement membrane to an avascular region of granulosa cells and back through the basement membrane into the blood stream. In t&o experiments to elucidate the follicular PREWULATCRY MONKEYS: MY of ESTROGEN SURGE (N=IS)

cell types responsible for the ovarian and peripheral venous oestrogen and progesterone and in vitro culture studies to assess the ability of the two cell types to synthesize both the steroids have been carried out in the rhesus monkey [18]. The ovary containing the large follicle secreted more oestrogen and progesterone into the ovarian vein blood than did the contralateral ovary containing no large follicles. If the large preovulatory follicles is removed, the estrogen and progesterone levels dropped to those observed in the ovarian vein draining the contralateral ovary. A day after the oestrogen surge, the levels of both oestrogen and progesterone in the ovarian and peripheral venous blood, dropped while that of progesterone in the follicular fluid appeared to be high (Fig. 2). This is the first observation of an increased progesterone level in follicular fluid of the rhesus monkey just prior mEwutAToRY MONKEYS: 1 MY ofmrE5lRoQEN SBGE (H5) Follicb-emmhin#

o#r

f9.2rO.e mm diomeIer1

,

Fig. 2. Levels of oestrogen and progesterone in peripheral plasm& ovarian venous blood and follicular fluid of rhesus monkeys on the day of oestrogen surge and 1 day after (24 h prior to ovulation). The values represent the mean f S.E. of the number of observations indicated at the top of the figure. Data taken from Channing, Wentz and Jones[SO] with permission.

783

Follicular maturation

AFTER REMDVAL DF FDLUCUUR

TIME

FLUID &ND

GRMULOSA

CaLs

b-616)

Fig. 3. Effect of removal of granulosa cells and follicular fluid on the levels of oestrogen and progesterone in ovarian vein blood at 5, 30 and 120min in rhesus monkeys. The levels are expressed as percentage of that prior to removal of granulosa cells and follicular fluid. The values represent the mean $- LE. of the number of observations given in-

side the bars. Data taken from Channing and Coudert[18] with permission. to ovulation. More recently, this has been confirmed using additional monkeys (Channing, manuscript in preparation). Removal of granulosa cells and follicular fluid from the large follicle had no effect on the levels of oestrogen and progesterone of ovarian venous blood after 5, 30 and 120min. (Fig. 3). These results indicate that under the experimental conditions employed, the granulosa cells of large follicles made no contribution to ovarian venous levels of oestrogen and progesterone. In these studies it could be concluded that the thecal cells are the principal source of steroids in the ovarian and peripheral venous blood. Similar findings have been made by Short[16] and Younglai and Short[26] in the mare. In isolated cell cultures, the thecal cells secreted more oestrogen, while the granulosa synthesized greater amounts of progesterone (Fig. 4). The ability of theta cells to secrete oestrogen and granulosa cells STEROIDOGENESIS

BY

MONKEY

to secrete progesterone has been studied in pregnant mare’s serum (PMS) treated monkeys (Fig. 5). Mixing of the two cell types or addition of exogenous testosterone to granulosa cell cultures resulted in greater secretion of oestrogen (Fig. 6). In culture or in short term incubations, granulosa cells have been shown to synthesize oestrogens if supplied exogenously with testosterone [27-321. These observations along with the earlier studies of Ryan et a/.[22 231 support a two-cell theory of intrafollicular oestrogen biosynthesis. But the in uiuo findings of Channing and Coudert[18] and the ultrastructural evidence of Crisp and Channing[25] and Meswerdt et a\[241 indicate that the thecal layer is the main source of oestrogen of ovarian and peripheral venous blood. Figure 7 illustrates diagrammatically this concept of follicular oestrogen biosynthesis. It is suggested that while thecal cells mainly contribute towards the circulating levels of oestrogen and progesterone, the granulosa cells may be responsible for the increased amounts of these hormones in the follicular fluid. Thanki and Channing[33,34] further observed that addition of oestrogen to porcine granulosa cell cultures led to a decrease in progesterone secretion. Thus, it is possible for follicular oestrogen along with other non-steroidal factors to prevent luteinization of granulosa cells prior to follicular rupture. (ii) LH/hCG receptors in granulosa Granulosa the ovarian of follicular ning[35,36]

ESTROCEN

cells offer a good test model for studying response to FSH and LH as a function maturation. Recently Sakai and Chanobserved that both small and large fol-

licles of the rhesus monkey contained

the same levels

of LH as measured by radioreceptor assay. Since all follicles are exposed to the same levels of LH, the differing response of them to mature in response to LH must lie in intraovarian mechanisms. The changes OVARIAN

CELL

THCCA

; d

cells

TYPES

IN CULTURE

GRANULOSA PROGESTEMNE

~;TROGEN

-5ZiF%Z

20,000 16.000 16.000 14.000 12.000

.

600

$

10.000

-

500

?j

6.000

.

400%

6.000

.

300

4,000

.

200

*

100;

2000 246610

3 ::

2466

lo DAVS

246610

w 5 =

24

IN CULTURE

Fig. 4. Levels of oestrogen and progesterone in cultures of thecal and granulosa cells harvested from rhesus monkeys and cultured for 8 to 10 days in medium 199 plus 15% monkey serum. The values represent the mean f SE. of the number of cultures shown on top of the diagram. Data taken from Charming, Wentz and Jones[sO] with permission.

784

G.

L.

KUMARI

and C. P.

CHANNING

STEROIDOGENSIS BY CULTURED MONKEY OVARIAN CELL TYPES HARVESTED FROM 6 PMSG-TREATED MONKEYS GRANULOSA ‘ESTROGEN

\ tONE.

PRDGESl

I3 1

THECA rESTROGEN

p 74000~ w 60,000 . it 50,000 x

40,000 -

2

30,000 -

!

0” 20,000 ” E I o*ooo Y! 2466

2466

24 6 DAYS IN CULTURE

8

Fig. 5. Levels of oestrogen and progesterone in cultures of thecal and granulosa cells harvested from 6 PMSG-treated rhesus monkeys and cultured for 8 to 10 days. Data taken from Charming, Wentz and Jones (1976)[50] with permission.

in follicular response to gonadotrophins has been examined in pig granulosa cells obtained from small, medium and large follicles. The granulosa cells of large follicles have IO-500 fold more LH/hCC receptors compared to those of small follicles [37-391. Charming and Kammerman[38] observed an increased num~r of LHjhCG receptors not only in the granuiosa cells, but also in the thecal cells as the follicle reached maturity. As the follicle matures there is ESTROGEN

SECRETION

BY MONKEY

an increase in the ability of the granulosa cells to accumulate cyclic AMP [40] and secrete progesterone Cl33 in response to LH. Granulosa cells obtained from small follicles are more responsive to FSH compared to those of large follicles in terms of their ability to bind FSH and stimulate cyclic AMP and progesterone levels (Fig. 8) [40,33] and Lindsey and Channing, unpublished observations). Nakano et a&41] reported similar findings.

OVARIAN CELL

TYPES

IN CULTURE GRANULOSA +

O.ISpg/ml

TESTOSTERONE 3opoo-

zi 25,000 e

-

GRANtLDSA

THiCA

Y i_= 20,000’ 2 ‘0 ,o

i!&ooo-

awlo,000

i!

GRANULOSA

THECA

% F

t

l-k I’b 2

4

6

6

IO

2

4

6

8 IO

~46610

,: 2

DAYS IN CULTURE

Fig. 6. Oestrogen secretion by thecal and granulosa cells obtained from three preovulatory monkeys and cultured alone, in combination or in the presence of O.l5fig/ml testosterone added each change of medium. The values represent mean + S.E. of the number of observations given at the top of the bar diagram. Data taken from Channing, Wentz and Jones[SO] with permission.

785

Follicular maturation

?MCA

f1Wli9WA -

Source of Estrogen

and Rogesterone

after ovulation

Fig. 7. Schematic diagram of follicular synthesis of oestrogen and progesterone. Channing, Wentz and Jones[SO] with permission.

(iii) Follicular jluid luteinization inhibitor

CHANGESIN LH AND FSH RESPONSIVENESS IN PORCINEGRANUWSP,CELLS OCCURRING DURING FOLLICULARMATURATION

E%

MEDIUM (I-Smm)

Data taken from

LARGE (6-12mm)

Fig. 8. Changes in LH and FSH responsiveness in porcine granulosa cells during follicular maturation. Data are taken from a Review by Channing, HillensjG and SchaerfCSl].

A compound(s) responsible for preventing luteinization of granulosa cells prior to ovulation is present in the follicular fluid [42-44]. Addition of 50% charcoal-treated follicular fluid obtained from small (l-2 mm) and medium (3-5 mm), but not large porcine follicles inhibited morphological luteinization and progesterone secretion by granulosa cells of large porcine follicles in culture (Table 1). This treatment also inhibited basal and LH-stimulated levels of cyclic AMP in porcine granulosa cells incubated for 24-48 h (Fig. 9). A similar inhibitor was found by Lunenfeld and his colleagues to be present in human follicular fluid [45]. Ledwitz-Rigby[43] also noted that the addition of methyl isobutyl xanthine, a phosphodiesterase inhibitor, reversed the inhibitory action of follicular fluid on cyclic AMP levels. It appears that the luteinization inhibitor reversed the inhibitory action of follicular fluid on cyclic AMP levels. Additionally, it appeari that the luteinization inhibitor prevents the stimulatory action of LH on levels of cyclic AMP and progesterone by interfering either with the adenyl cyclase or cyclic AMP, but not by blocking LH/hCG receptors (Anderson and Channing, unpublished observations). Induction of LH/hCG receptors is due to FSH action upon the follicle [2]. Channing and Anderson have observed

Table 1. Influence of follicular fluid on progestin secretion during 48 h incubations (mean + SE.) Progestin secretion (ng/5 x 10’ cell/day) Type of culture

Medium + 50% SFFI

Medium + 50% LFFI

1

Monolayer

120,000

2

Monolayer

5

Suspension

6

Monolayer

4,700 rl: 400 (3) 1,100 * 43 (3) 5,656 k 548 (4) 423 + 7 (3)

Expt

2,324 f 90 (3) 40,121 f 2,167 (4) 10,595 k 429

Other

2,459 + 156(2) (50% serum) 18,972 + 4,620(6) (medium 199D)

Granulosa cells obtained from large (612 mm) porcine follicles were incubated for 48 h as monolayers or suspensions. Follicular fluids and Sera were pretreated to remove endogenous progesterone. Data were taken from Ledwitz-Rigby et al. [44]. SFFI = Follicular fluid of small follicles; LFFI = Follicular fluid of large follicles.

G. L. KUMAN and C. P.

786 %

LARGE FFL

too 100

z

CHANNING

IOOO-

62 75

T

39 57

0

CONTROL

m

l.Op.g/ml

oLH DURING

POST

INCUBATION

’ 80007 i : 600“2 :: : 5 0 I0

62 69

400200-

I

50%

LARGE FFL

50%

MEDIUM FFL

50V. PIG SERUM

50% SMALL FFL

48 HOUR PRElNCU6ATlON

CONDITIONS

Fig. 9. Effect of porcine follicular fluid (FFL) of small (l-2 mm), medium (35 mm) and large (612 mm) follicles and serum on accumulation of cyclic AMP in granulosa cells of large porcine follicles with or without LH. Granulosa cells were preincubated for 48 h with SO”/, follicular fluid and the cells washed and reincubated for 30min after adding 1 &ml ovine (0) LH. Intracellular levels of cyclic AMP were measured.

follicular fluid which both during follicular ation, and prior to ovulation, may overcome hibitor. The stimulator(s) may be under the of FSH (Anderson and Channing, unpublished vations).

that the luteinization inhibitor present in small follicular fluid can inhibit FSH induction of LH/hCG receptors in cultured granulosa cells obtained from small porcine follicles [Z]. It is possible that atresia of the follicles and the simultaneous choice of which follicle will mature may be related to the amount of luteinization inhibitor present in the follicular fluid. Since large follicles do not contain luteinization inhibitor, Ledwitz-Rigby et aI.[44] proposed the probable presence of a luteinization stimulator(s) in the

CONTROL

LUNG

HEART

GRANULOSA CELLS

LUNG

binding inhibitor in the corpus luteunl

In most lower mammals, granulosa cells form the bulk of the corpus luteum whereas in the primates, thecal cells also form a significant portion of the

a

m

IID

HEART

(iv) LH/hCG

GRANULOSA

0

m

m

%k:y’

CORPUS LUTEUM (CL)

NON LUTEAL TISSUE (NLI

WHOLE OVARY

r

-II

I IO

2 ;_

100 90

i

60

x -

70 60

d

50

f B

40

u

30

&

20

$

10 CONTROL

47.0

33.0

2.0

O.?

2-Y

WEEKS

maturthe incontrol obser-

0.7

TISSUE

1.9

3.5

STORED

7.0

9.0

9.0

11.0

11.0

-2O*

Fig. 10. Ability of the 30,OOOy supernatant fractions of aqueous extracts of porcine tissues stored for several weeks at -35°C to inhibit binding of ‘?-hCG to porcine granulosa cells of large follicles. The binding is expressed as percentage of control binding and the values represent the mean f S.E. of 5 observations. The duration of storage of tissues is shown at the bottom. Data taken from Sakai et al.[47].

787

Follicular maturation corpus luteum. Local control of corpus luteum function and responsiveness of follicles to gonadotrophins may be under the influence of a LH receptor binding inhibitor (LHRBI). Yang, Samaan and Ward[46] observed that extracts of frozen pseudo-pregnant rat ovaries have the ability to inhibit LH binding to luteal cell receptors. Later, Sakai et a/.[473 found a similar inhibitor in the frozen aqueous extracts of pig corpus luteum, but not in non-luteal ovarian tissues, lung or heart (Fig. 10). Inhibition of the binding of “‘1-hCG to granulosa cells of large porcine follicles is observed with the supernatant of the 30,OOOyfraction after centrifugation of the aqueous extracts of pig corpora lutea obtained from pigs at the early, mid, and late luteal phases of the cycle. Maximum inhibition of hCG binding was brought about by extracts of late and mid cycle luteal extracts. Extracts of early corpus luteum had a low ability to inhibit hCG binding [48] (Fig. 11). Addition of 10% aqueous extract of the 30,OOOy supernatant fraction from corpus luteum harvested during the late luteal phase (old corpus luteum) to cultured porcine granulosa cells obtained from large follicles, either directly or after removal of steroids with 0.1% charcoal (w/v), inhibited progesterone secretion in O-2 day and 24 day cultures. Simultaneous addition of ovine LH (1 pg/OS ml) failed to reverse the inhibitor effect on progesterone levels [49] (Figs. 12 and 13). The increase in LHRBI activity as the corpus luteum ages may be responsible for the regression of the corpus luteum at the end of a non-pregnant estrous cycle. Its inhibitory effect on progesterone secretion by granulosa cells of large follicles suggests that LHRBI

may diffuse to adjacent follicles and control responsiveness.

their

CONCLUSIONS The selection of the primary follicle, which will undergo maturation and ovulation, may be dependent on the changes in the ovary whereby follicular responsiveness to LH and FSH are altered. The mature follicle attains more LH receptors, compared to the small and medium follicles which respond more to FSH. The follicle thus destined to ovulate will secrete oestrogen and progesterone into the circulation, from the thecal cells which are the principal source of the steroids. Granulosa cells also synthesize oestrogen if supplied with exogenous androgen, which may contribute towards the follicular fluid levels of oestrogen prior to ovulation. Luteinization of granulosa cells of large follicles can occur spontaneously if they are removed from their follicular fluid environment. Follicular fluid from small and medium follicles contains a luteinization inhibitor which blocks morphological luteinization and LH-stimulated cyclic AMP and progesterone levels in granulosa cells from porcine follicles. The inhibitor regulates follicular responsiveness to gonadotrophins by inhibiting FSH induction of LH receptors and perhaps preventing some follicles from maturing. The presence of an LH binding inhibitor in corpus luteum and the observation of its increased activity as the organ ages, suggests an important role for this inhibitor in the regulation of corpus luteum function

C

0 M Y L H

CONTROL OLD LUTEAL MID-LUTEAL EARLY LUTEAL LUNG HEART

Fig. 11. Ability of the 30,000 g supernatant aqueous fractions of extracts of luteal and non-luteal ovarian tissues obtained during the late (old), mid and early luteal phases of the estrous cycle, lung and heart to inhibit binding of “‘I-hCG to granulosa cells of large follicles. The values represent the mean f S.E. of 5 observations. The open bars show addition of extracts without prior charcoaltreatment and dotted bars represent addition of extracts after treatment with 0.1% charcoal (w/v) c491.

G. L. KUMARIand C. P.

c =

CHANNING

CO&TROL

0 = OLD LUTEAL A( = WIR.LUAL Y EARiY L.!IT~AL q

!

FL T

!H .

l

Y

;



tnorcoa/

hated

CL

Y

04:

UkrCool hated cd.

Fig. 12. Effect of addition of 10% of the 3O.OOOysupernatant fractions of extracts of corpus luteum harvested from the late (old), mid and early luteal phase on progesterone secretion by 0 to 2 day cultures of granulosa cells harvested from large porcine follicles cultured with or without 1 &ml ovine LH. The luteal extracts were added either before or after treatment with 0.1% charcoal (w/v). The values represent the mean k S.k. 01 Y cultures taken from three individual experiments. a = P < 0.001: b = P i 0.005 vs control

450 r\ g: 400 C .. 3 350 t1 300 VI Y 250 3 $ 200 0’ 4 $ 150 Y 2

100

2 3::

50

z

0

cffpu5 hJfwm extract

Fig. 13. Elfect of addition of 10% of the 30,OOOgsupernatant fractions of extracts of corpus luteum obtained from late (old), mid and early luteal phase of the porcine estrous cycle upon progesterone secretion by cultures of granulosa cells obtained from large follicles cultured from day 2 to 4. Some of the luteal extracts were added either before or after treatment with 0.1% charcoal (w/v). The values represent the mean f S.E. of 9 observations of three individual experiments. C = control; 0 = old (late); M = mid-luteal;. Y = early luteal; a = P < 0.001; c = P < 0.01; d = P < 0.05 vscontrol.

as well as control of adjacent follicles in their response to LH. A delicate balance among the steroidal and non-steroidal factors may be responsible for the maturation and selection of a dominant follicle deatined to ovulate and form the corpus luteum.

REFERENCES

I Channing C. P. and Tsafriri A.: Mechanism of action of luteinizing hormone and follicle stimulating hormone on the ovary in vitro. Meraholism 26 (1977) 413468.

Foilicular maturation 2. Channing C. P., Anderson L. D. and Batta S. K.: Follie&u growth and development. C&tics in Obstetrics and Gyaecotogy (Edited by J. E. Tyson), Vol. 5 (1978) pp. 333-390. 3. Schwartz N. B.: A model for the regulation of ovulation in the rat. Recent Prog. Harm. Res. 2S (1969) I-55. 4. Lindner H. R., Amsterdam A., Sabmon Y., Tsafriri A., Nimrod A.. Lamprecht S. A., Zor U. and Koch Y.: intraovarian factors in ovulation: determinants of follicular response to gonadotrophins. J. Reprod. Frrr. 51 (1977) 215-235. 5. Channing C. P.: Effects of stage of the menstrual cycle and gon~otropins on luteinization of rhesus monkey granulosa cells in culture. Endocrinology 87 (1970) 49-60. 6. Charming C. P.: Temporal effects of LH, hCG, FSH and dibutyryl cyclic 3’S’-AMP upon luteinization of rhesus monkey granulosa cells in culture. E~ocr~~logy 94 (1974) 1215-1223. 7,* Channing C. P.: Influences of the in viuo and huitro hormonal environment upon luteinization of granulosa cells in culture. Recent Prog. i-form. Res. 26 (1970) 589-622. 8. Channing C. P.: Steroidogenesis and morphology of human ovarian cell types in tissue culture. J. Endocr. 45 (1969) 297-308. 9. McNatty K. P. and Sawyers R. S.: Relationship between the endocrine environment within the Graafian foilicle and the subsequent rate of progesterone secretion by human granulosa cells in vitro. J. Endocr. 66 (1975) 391-400. 10. Batta S. K., Wentz A. and Jones G.: Effect of testosterone on estrogen and progesterone secretion by human granulosa and granulosa plus thecal ceil cultures. Abstract 51. In the Proc. 10th Annual Meeting of the Society of Reproducrion. Austin. Texas. August 14-17, 1977. 11. Batta S. K., Channing C. P., Wentr A. and Jones G. S.: Influence of follicular maturation and testosterone upon estrogen and progesterone secretion by human follicular cell types cultured in oitro (manuscript submitted). 12. Charming C. P. and Seymour J.: Etfects of dibutyryl cyclic 3’,5’-AMP and other agents upon luteinization of porcine granulosa cells in culture. Endocrinology 87 (1970) 165169. 13. Channing C. P.: Effect of stage of the estrous cycle and gonadotrophins upon luteinization of porcine granulosa cells in culture. ~~oeri~~ogy 87 (1970) 150-164. 14. Goldenberg R. L., Bridson W. E. and Kohler P. 0.: Estrogen stimulation of progesterone synthesis by porcine granulosa cells in culture. Biocfiem. Biophys. Res. Commun. 48 (1972) 101-107. 15. Van Thiel D. II.. Bridson W. E. and Kohler P. 0.: Induction of “luteinization” in cultures of granulosa cells by chorionic gonadotropins. Endocrinology 89 (1971) 622-624. 16. Short R. V.: Ovarian steroid synthesis and secretion in vioo. Recent Prog. Horm. Res. 20 (1964) 303-323. 17. Resko J. A., Koering M. J., Goy R. W. & Phoenix C. H.: Preovulatory progestins: observations on their source in monkeys. J. cfin. Eadocr. Metab. 41 (1975) 120-125. 18. Channing C. P. and Coudert S. P.: Contribution of granulosa cells and follicular fluid to ovarian estrogen secretion in the rhesus monkey in ho. Endocrinology 98 (1976) 590-597. 19. Strott C. A., Yoshimi T., Ross G. T. and Lipsett M. B.: Ovarian Physiology: Relationship between plasma LH and steroidogenesis by the follicle and corpus luteum; Effect of hCG. J. c/in. Endocr. Metah. 29 (1969) 1157-1167.

789

20. Mikhail G.: Sex steroids in blood. Clin. Obstet. Gynecot. 10 (1967) 29-39. 21. Falck K. B.: Site of production of oestrogen in rat ovaries as studied by micro transplants. Acta Physiof. &and. (SuppI) 47 (1959) 163-193. 22. Ryan K. J., Petro 2. and Kaiser J.: Steroid formation by isolated and recombined ovarian grant&a and thecal cells. J. Cfin. Endoer. Metab. t8 (1968) 355-358. 23. Makris A. and Ryan K. J.: Progesterone, androstenedione, testosterone, estrone and estradiol synthesis in hamster ovarian follicle cells. Endocrinology 96 (1975) 694-701. 24. Meswerdt W., Muller 0. and Brandau H.: Die differenzierte structur und funktion der granulosa und theka in verschiedenen follikel-stadien menschlicher ovarien. Arch. Gynak. 222 (1977) 45-71. 25. Crisp T. M. and Channing C. P.: Fine structural events correlated with progestin secretion during lutein~ation of rhesus monkey granulosa cells in culture. Biof. Reprod. 7 (1972) 55-72. 26. Younglai E. V. and Short R. V.: Pathways of steroid biosynthesis in the intact Graafian follicle of mares in o&trus. J. Endoer. 47 (1970) 321-331. 27. Rvan K. J. and Short R. V.: Formation of estradiol by granulosa and thecal cells of the equine ovarian follicle. Endocrinology 76 (1965) 108-l 14. 28. Channing C. P.: Studies on tissue culture of equine ovarian cell types: pathways of steroidogenesis. f. Endocr. 43 (1969) 403414. 29. Erickson G. F. and Ryan K. J. : The effect of LH/FSH. dibutyryl cyclic AMP and prostaglandins on the production of estrogens by rabbit granulosa cells. Endoe~i~o~o~~97 (1975) 108-l 13. 30. Moor R. M.: Sites of steroid production in ovine Graafian follicle in culture. J. Endow. 73 (1977) 143150. 31. Fortune J. E. and Armstrong D. T.: Androgen production by theta and granulosa isolated from proestrus rat CollicIes. E~ocri~Iffgy 108 (1977) 1341-1347. 32. Fortune 3. E. and Armstrong D. T.: Hormonal control of 17&estradiol biosynthesis in proestrus rat follicles: Estradiol production by isolated theta versus granulosa. Endocrinoloav 102 (1978) 227-235. 33. Thanki K. H. andChan&ng C. P.: Influence of serum, estrogen and gonadotropins upon growth and progesterone secretion by cultures of granulosa cells from small porcine follicles. Endocr. Res. Commun. 3 (1976) 319-333. 34. Thanki K. H. and Charming C. P.: Effects of folliclestimulating hormone and oestradiol upon progesterone secretion by porcine granulosa cells in tissue culture. Endocrinology 103 (1978) 74-80. 35. Sakai C. N. and Channing C. P.: Evidence for uptake of biologically active luteinizing hormone (LH) into the preovulatory monkey follicle in t&o. Abstract presented at the Meeting of the Federation of American Society of Experimental Biology held at Atlantic City, April 1978. 36. Sakai C. N. and Channing C. P.: Evidence for uptake of radioreceptor-~sayable luteinizing hormone into the preovulatory monkey follicle in viw. Endocrinology (1979) In press. 37. Channing C. P. and Kammerman S.: Characteristics of gonadotropin receptors of porcine granulosa cells during follicle maturation, E~ocri~~ogy 92 (1973) 531-540. 38. Channing C. P. and Kammerman S.: Binding of gonadotropins to ovarian cells. Biol. Reprod. IO (1974) 179-198. 39. Kammerman S. and Ross J.: Increase in numbers of gonadotropin receptors on granulosa cells during follicle maturation J. c/in. Endocr. Mefah. 41 (1975) 546-550. 40. Lindsey A. M. & Channing C. P.: Influence of follicu-

790

41.

42.

43.

44.

G. L. KUMARIand C. P. CHANNING lar maturation on the ability of porcine granulosa cells (GC) to produce cyclic AMP in response to LH and FSH. Abstract presented at the 8th Annual Meeting of the Society for the Study of Reproduction held at Philadelphia,- August 11-14, 1976. _ Nakano R.. Akahori T.. Katavama K. and Toio S.: Binding of.LH and FSH to porcine granulosa cells during follicular maturation. J. Reprod. Fert. 51 (1977) 23-27. Bernard J.: El&t du liquide folliculaire sur la luteinization in vitro des cellules granulosaires du rat. Comptes Rendus des Secencesde la Societe de Biologic et de Sa Hiales. 167 (1973) 882-885. Ledwi&Rigby F.:‘Possible inhibitory site of action of porcine follicular fluid upon granulosa cell luteinization. Abstract presented at the 7th Annual Meeting of the Society for the Study of Reproduction held at Ottawa 1974. Ledwitz-Rigby F., Rigby B. W., Gay V. L., Young J., Stetson M. and Channing C. P.: Inhibitory action of porcine follicular fluid upon granulosa cell luteinization: Assay and influence of follicular maturation. J. Endocr. 74 (1977) 175-I 84.

45. Lunenfeld B. I., Ben-Aderet N., Michael R. B., Griistein S., Kraiem Z.. Potashak G., Shalit A., Tikotzky D. and Rofehh G.: Correlation of hormonal profile and ovarian morphological features during the preovulatory period in the human. In Endocrinology of the Ovary (Edited by R. Scholler) Proceedings of a Conference held in Fresnes, France 1976. Editions Sepe, Paris (1978) pp. 187-202.

46. Yang K. P., Samaan N. and Ward D. N.: Characterization of an inhibitor for luteinizing hormone receptor site binding. Endocrinology 98 (1976) 233-242. 47. Sakai C. N.. Enael B. and Channina C. P.: Ability of an extract of pig corpus luteum to inhibit binding of ‘2sI-labelled human chorionic gonadotropin to porcine granulosa cells. Proc. Sot. Exp. Biol. Med. 155 (1977) 373-376. 48. Tucker S., Kumari G. L. and Channing C. P.: Observation of greater LH/hCG binding inhibitor activity in aqueous extracts of old compared to young porcine corpus luteum. In Ovarian Follicular and Corpus Luteum Function (Edited by C. P. Channing, J. Marsh and W. Sadler). Plenum Press, New York, 1978 (in press). 49. Kumari G. L., Tucker S. and Channing C. P.: Demonstration of a larger amount of inhibitor of binding of labelled human chorionic gonadotropin and of progesterone secretion by cultured porcine granulosa cells in aqueous extracts of old compared to young corpus luteum, 1978 (submitted for publication). 50 Channing C. P., Wentz A. C. and Jones G. S.: Steroid secretion by human and monkey ovarian cell types in vivo and in vitro. In Symposium on the Ovary (Edited by R. Scholler) held at Fresnes, France 1976, pp. 71-86. Editions Sepe, Paris (1978). 51 Channing C. P, Hillensjb T. and Schaerf F.: Hormonal control of oocyte meiosis, ovulation and luteinization in mammals. In Clinics in Endocrinology and Metabolism (Edited by M. Lipsett and G. Ross). Saunders, Philadelphia, Vol. 7 (1978) pp. 60624.

Intraovarian control of progesterone biosynthesis by granulosa cells and corpus luteum.

I I. pp. Journal of Srrraid Biochemisrr): Vol. 781 to 790 Per&w~~on Press Ltd 1979. Printed in Great Britain INTRAOVARIAN BIOSYNTHESIS CONTROL OF P...
934KB Sizes 0 Downloads 0 Views