Testicuiar steroidogenesis in the baboon papio anubis

187

2305

TESTICULAR

James Department The

IN THE BABOON PAP10 --

STEROIDOGENESIS P.

Preslock

of

Reproductive

University

of

Emil

and

Texas

Steinberger

Medicine Medical

ANUBIS

and

School

Biology

at

Houston

and Program The

in

University

Received

of

Reproductive Texas

Biology

Graduate

School

Houston,

Texas

and

Endocrinology

of

Biomedical

Sciences

)+-14-78 ABSTRACT

converts pregnenolone We recently reported that the baboon testis studies were to testosterone through the delta-4 pathway. The present de1 tato determine the metabolism of intermediates of the delta-4 and Fragments (50 mg) were incubated for 3 5 pathway by the baboon testis. pregnenohr with 10 nCi of the following tritium-labelled substrates: lone, progesterone, 17-hydroxypregnenolone, 17-hydroxyprogesterone, Pregnenolone dehydroepiandrosterone, androstenedione, or testosterone. the delta-4 pathway, was converted to testosterone primarily through progesterone, 17-hydroxyprogesterone and 20awith accumulation of dihydroprogesterone as predominant intermediates. Similar results were obtained in progesterone incubations. 17-Hydroxyprogesterone was not efficiently metabolized by the fragments, while 17-hydroxypregconverted nenolone and dehydroepiandrosterone were efficiently into testosterone and androstenedione. Androstenedione was metabolized primarily to testosterone, while testosterone was not a suitable substrate. Sane 5u-androstanediol was identified in each incubate. These results suggest that al though testosterone is formed from the delta-5 intermediates pregnenolone through the de1 ta-4 pathway, the baboon are more suitable substrates for testosterone synthesis in testis.

The vert

of

the

pregnenolone

pathway. lone

testis

to

This

pathway

subhuman

to

testosterone of

conversion

progesterone,

with

17_hydroxyprogesterone,

to

VoZwne

32,

Number 2

primate

a

S

has

been

primarily involves

further

androstenedione,

TDEOXDI

demonstrated through

the

metabolism

metabolism and

of then

to

of

con-

de1 ta-4 pregneno-

progesterone to

to

testosterone.

September,

2978

This

pathway

has

mosets

Saguinus

baboon

Papio

Macaca

mulatta

In

our

conversion ments.

the

studies

anubis

of

studies

was

(41,

pregnenolone

However,

we did enzymes

delta-4

or

the

therefore

steroidogenic

establish

the in

and

and

as

predominant

Callithrix

earlier

in

jacchus

reported

in

the

(3),

the

mar-

in

Rhesus

the

monkey

(5).

earlier

were

reported

(1,2)

(4),

testicular

synthesis

recently

Oedipus

steroidogenic of

been

to not

by

testosterone

determine

investigating

delta-5

pathways

undertaken enzymes

delta-4

this

we demonstrated

pathway

subhuman

primate

to in as

a

by

predominant

baboon

the

specificity

the

metabolism

by

baboon

Papio

testicular of of

testes.

investigate anubis,

f ragtesticular

intermediates The

the

predominant

de 1 ta-4

following

specificity and

for

to

of further

testosterone

specie.

MATERIALS AND METHODS Testicular

Tissues:

Testes were removed from mature baboons (Papio anubis) and placed were weighed, into ice-cold tris-sucrose buffer, pH 7.4. The---- testes and cut into approximately equal 50 mg fragments. The decapsulated, fragments were teased apart and placed into flasks for incubation studies. Materials: silica-gel (Silicar (nanograde 1 and Solvents Louis; purchased from Mallinckrodt Chemical Works, St. Wil ton, steroid carriers were ob ained from Steraloi f tracers cPaC) were Radioactive substrates ( Ii) and Amersham Searle, Inc., Arlington Heights, Ill. Cofactors incubations were obtained from Sigma Chemical Co., St. purchased 1) for chranatographic separations was (No. Steroids were checked for purity Company. Paper chromatography, while paper and silica gel were washed prior to use in these studies.

were TLC- 7Gf ) nonradioactive New Hampshire. purchased from utilized for Louis. Paper from Whatmann thin-layer by with

methanol

S

-lFDEIEOXDrn

Incubations: Testicular fragments were placed into incubation flasks containing Krebs-Ringer bicarbonate buffer, pH 7.4, fortified with NADH, and an NADPH generating system which contained NADP, nicotinamide, glucose6-phosphate, pyruvate, glucose-6-phosphate dehydrogenase, and lactic dehydrogenase. The fragments were incubated for 3 hr with 10 uCi of the following tritium-labelled substrates: pregnenolone, li'-hydroxypregnenolone, dehydroepiandrosterone, progesterone, 17-hydroxyprogesterone, androstenedione, and testosterone. Each substrate was incubated in triplicate for a total of 21 separate determinations. Incubations were performed in a Dubnoff metabolic shaking incubator at 37'C under an atmosphere of 95% 0 15% CO . Upon completion of the incubations the reactions were teriinated sith 0.5 ml 1N HCl, and the incubates were immediately frozen. Extractions: The incubates were thawed and the testicular fragments were homogenized in Krebs-Ringer bicarbonate buffer pli 7.4. The homogenized fragments were pooled with their Radiolabelled tracers (14C) and ,lab~~E~~ct~,",rie~~u~~~onadd~~di~~ each pool, and the pools were extracted ten times with cold diethyl ether:chloroform, 4:l. The solvents were evaporated with nitrogen, and the residues were concentrated. M'ethanol (5 ml) was added to each tube and aliquots were removed for estimates of recoveries. The remaining solvent was evaporated under nitrogen, and the residues were placed onto paper strips for chromatographic separations. Chromatographic procedures: The residues were placed onto Whatmann No. 1 paper strips (2.5 x 50 cm) which were impregnated with formamide. The strips were chromatographed in hexane, with a second chromatographic separation in a hexane:benzene mobile phase. Upon completion of the chrcmatographic separations, the paper strips were dried in a warm air oven. The radioactive peaks were located with a Packard Model 385 Recording Ratemeter. The peaks were eluted from the paper strips with 80 ml of methanol, and the metabolites were isolated by thin-layer chranatography in selected solvent systems. The systems utilized in these studies were benzene:ethyl acetate (80:20, 60:40), benzene:methanol (98:2, 95:5), and chloroform:acetone (90:10, 80:20). Testosterone, 2Oa-dihydroprogesterone, 5a-androstanediol and dehydroepiandrosterone were acetylated, and were separated by thin-layer chromatography. Aliquots for recovery determinations were obtained prior to each thin-layer separation and prior to crystallization of each metabolite. Crystallization procedures: Following isolation and tentative identity by thin-layer chromatography, final identity of each metabolite was es5abi&shed by crystallization to constant specific activities and HI C ratios through

three successive solvent combinations (acetone:hexane; acetone:cyclohexane; acetone: hexane). Conversion qf substrates to metabolites was calculated by determining the total H-DPH for 3each metabolite from crystallization data, correcting the t tal H-DPM for procedura 1 q and dividing the corrected total losses,3 H-DPM for each metabolite by the H-DPM of thq total incubate. Conversions were expressed as a percentage of the 5a-Androstanediol H-DPM of the total incubate. and 2Oa-dihydroprogesterone, although not crystallized, were identified by acetylation and comparison of relative mobilites in four separate thin-layer chranatographic Percent sys terns. conversion of each substrate into these metabolites was estimated from recovery the data. RESULTS Baboon to

testosterone

gesterone, the of

testicular

primarily

metabolites

respective

delta-5

the

was

incubates,

was

1.1%

while

A as

( 0.8%)

were

the

was

non-metabolized

pregnenolone

pathway

(Fig.

30,5%,

minor

in

the

radioactivity 1.5%.

identified

as

pregnenolone

Pro-

in

Sane

and

of

was

24.2%

through

the

(0.9%)

and

incubates. the

occurred

radioactivity,

the

14.8%

Tes-

pregnenolone

5a-reduction

2.4%

substrate

were

19.32,

conversion

identified

was

1).

20a-dihydroprogesterone

17-hydroxypregnenolone

total

androstenedione

5a-androstanediol

delta-4

representing

apparent

of

radiolabelled

and

radioactivities.

pathway

tosterone

while

through

identified,

dehydroepiandrosterone

as

converted

17-hydroxyprogesterone,

major the

fragments

of

the

incubate

radioactivity. Similar since

results

progesterone

were

while

was

(36.3%) 1.1%

of

the

5a-androstanediol

demonstrates incubations.

crystallization

in

progesterone

incubates

17-hydroxyprogesterone

(29.5%),

dihydroprogesterone tosterone

obtained

were

the

major and

radioactivity

was

4.3% data

of

the for

total

(23.7%)

steroids

11,

and

2Oo-

identified.

androstenedione radioactivity.

pregnenolone

(Fig.

and

Teswas

1.9%,

Table progesterone

1

191

Figure

Percent (X) conversion of ptegnenolone and progesterone to metabolites by baboon testicular fragments. The following abbreviations are used in the figures: PRR, pregnenolone; PRO, progesterone ; 17-PRO, S7-hydroxy20a-dihydroprogesterone; progesterone; 20a-PRO, 17-PRR, 17-hydroxypregnenolone; DEA , dehydroepiandrosterone; AND, androstenedione; TRS, testosterone; 5a, 5a-androstanediol.

1.

A substantial remained

(5.7%)

of

for

of

testosterone

Incubations conversion

of

with of

this

the

with

synthesis

occurred

pathway by

of

Table

2).

androstene-

the delta-4

more

suitable

testicular

resulted

testosterone

(86.5%)

in the incubates.

were

baboon

2;

C?.OX) f

intermediates

17-hydroxypregnenolone into

(Fig.

to testosterone (3.8%)

delts-5

substrate

substrate

the 3 hr incubation

substrate

to incubations

intermediates

strates

this

17-hydroxyprogesterone

and 5a-androstanediol

In contrast way,

of

at the termination

Some conversion dione

portion

in and

pathsub-

fragments. a

substantial

androstenedione,

a

TES AND 5a

3,360 2,670 3,540 118 210 474

---149 il0 160 5 10 22

Z 6 8 13

164 99 128 86

3H DPM/III~~'~

48 3 4 7

1.8 1.3 1.6 2.1

1.6 1.1

3 9

91 97

106 90

1.5 1.1

1.6 1.1

3R/'4C

___II

14C DPM/mge'f

2,970 2,190 3,200 103 195 435

3,270 1,970 2,560 1,730 100 84 118 169 264

3H DPM (T)g

^ 88 82 90 87 93 92

91 92 90 93 92 89 94 91 92

% Pur&

% Purity = 3R DPH (T)/3R DPM (I),

17-PRO, 17-hydroxyproThe following abbreviations are used in the tables: PRO, progesterone; gesterone; 2&r-PRO, ZOC-dihydroprogesteron~; PRE, pregnenolone; If-PRE, 17-hydroxypre%~en~lone; DEA, dehydroepiand~oster~ne; TES, testosterone; AND, androstened~one~ 5a, Sa-androstanediol. Approximately 20 mg of crystalline steroid was added to each sample prior to crystallization. ~on~~ta~oliz~d subgtrate. Initial H-DPM (Xl.0 )3before crystallization. Data as mean DPM (XI0 ) of three successive crystallizations of duplicate samples. Solvent systems for crystallizations were: 1. Acetone/Hexane; II. Acetone/Cyclohexane; III. Acetone/ Nexane3 Total H DPM (X~03) following crystallization,

2oa-PRO

PRO 17-PRO

FFo~esteroneYZFstrate

.-~_I___~

Crystallization $ata for metabolism of pregnenolor~-7-~H (10 pCi> and progesterone-7- H (10 uCi) by baboon testicular fragments (Papio anubis) at 3 hr of incubation. ---

2:

3H DFM (T)d Metabolite s,b Prennenolane substrate PRO_ 3,580 17-PRO 2,140 2,840 20a-PRO PREC 1,610 17-PRE 108 DE#i 95 126 TES AND 186 286 5iY

~-~-

TABLE

S these

representing

metabolites

radioactivities

(Fig.

17-hydroxyprogesterone identified

into

substrate

remained

Figure

(0.4X),

2.

metabolites

of

the

respective

Dehydroepiandrosterone

(0.3X),

(3.0%)

17-Hydroxypregnenolone

was

by the baboon of

testis

as only

0.3%

were readily of

this

the incubations.

(X) conversion of 17-hydroxyprogesterone and 17Percent tes titular hydroxypregnenolone to metabolites by baboon Figure 1 lists metabolite abbreviations. fragments.

terone

and androstenedione

of

total

androsterone

62.1%

5a-androstanediol

and

upon termination

Dehydroepiandrosterone

the

and

2).

metabolites.

as minor

converted

28.2%

Table

2,

193

?EBEOXDb

was also

a suitable

substrate

these

metabolites

were

27.9%

( 1.3%)

and

as

radioactivity. substrate

5a-Androstanediol. (2.5%)

were also

identified

for and

testos59.8%

dehydroepi-

in the incubat.es.

a,b

8

h

substrate 42 32 3,570 7,870 355 42

substrate 8,960 209 785 301

311 DPM (Xld

2 2 167 364 16 2

407 11 35 14

3 H DPPf/mge,f

1 92 302 2

1.2

415 7 29

3H/ 14C

1.4 1.8 1.2

.9 1.6 1.2

14 C DPM/mge,f

37 31 3,340 7,280 321 37

8,130 230 706 280

3H DPM (Tjg

Crystallization data for metabolism of 17-hydroxyprogesterone-73W (10 nCi1 and 17-hydroxypregnenolone (10 uCi) by baboon testicular fragments (Papio anubis) at 3 hr of incubation --

Table 1 lists metabolite abbreviations. Approximately 20 mg of crystalline steroid was added to each sample prior to crystallization. Non-meta olized subgtrate. $ Initial H DPM (X10 l3before crystallization. Data as mean DPM (Xl0 ) of three successive crystallizations of duplicate samples. Acetone/Hex~~; II. Acetone~Cyclohe~~ne; System for crystallizations *i-e: I. Solvent Acetong/Nexane. crystallization. Total W D9 (X1031 fojlowing X Purity = N DPM CT)/ H DPM (I).

-l7-Hydroxypreg~~~lone 17-PRl?F DEA TRS AND 5a 17-PRO

5a

17-Hydroxyprogesterone 17-PRO’ Tl?s AND

Metabolite

TABLe 2:

87 94 94 93 90 89

91 91 90 93

III.

Purityh

s A substantial strate

portion

was converted

of

into

portion

of

the substrate

period.

5a-Androstanediol

195

lCBEOXDIll

the

radiolabelled

testosterone (5.8%)

(86.9X),

remaining

was 2.4%

androstenedione

of

at

with

anly

the end of

the

total

the

suba

small

incubation

radioactivity

from

androstenedione. Testosterone fragments

since

incubation

g7.iX

period.

converted (1.4%).

was not a suitable

into Figure

of

testosterone

However,

3.

a

androstenedione 3 and Table

andros terone ) androstenedione

Figure

substrate

3

for

baboon

testicular

remained

at

the

portion

of

testosterone

small (4.9.%),

and

demonstrate and testosterone

the

into data

end

of

the was

So-androstanediol for

dehydroepi-

incubations.

Percent (%I conversion of dehydroepiandrosterone, stenedione and testosterone to metabolites by testicular fragments. Figure 1 lists metabolite viatians.

androbaboon abbre-

339 36 15

607 34 17

Androstenedione substrate TBS ANDC 8,850 825 5ci 319

Testosterone substrate TESC 13,800 AND 741 5u 380 1.6 1.1

1.6 1.0

361 31

211 34

8

1.3 ___--_I_--

73 234

--

3H/"4C

1.6 1.0

-

---p14 C DpM/mg e,f

12,100 680 343

7,980 724 302

2,440 4,880 114 211 _p--

__--

3H DPM (T)'

W-P

III.

88 92 90

90 88 95

94 92 91 89

% Purityh

Table 1 lists metabolite abbreviations. Approximately 20 mg of crystalline steroid was added to each sample prior to crystallization. Non-meta olized subgtrate. 9 Initial H-DPM (X10 j3before crystallization. Data as mean DPM (X10 ) of three successive crystallizations of duplicate samples. II. Acetone/Cyclohexane; Solvent systems for crystallizations were: I. Acetone/Hexane; Acetone/Hexane. Total 3H DPM (X103) following crystallization. % Purity = 3H DPM (T)/~H DPM (I).

122 244 6 11

3H DPM (IId

3 H DPM/mg e,f

D~bydroepiandrosterone substrate TBS 2,610 AND 5,300 5o 126 DEAC 237

Metabolite a,b

TABLE 3: Crystallipation $ata for metabolism of dehydroe~ia*d~osterone-7-3H (10~ Ci), androstenedione-7- H (10 PCi) and testosterone-7- H (10~ Ci) by baboon testicular fragments (Papio anubis) at 3 hr of incubation. --

_DISCUSSION results

The

the baboon

of

Papio

marily

through

bolism

of

the delta-4

are

pathway.

to

similar

Saguinus

to recent

Oedipus -.

Th is

testis

of

testosterone

pri-

involves

meta-

17_hydroxyprogesterone,

with

progesterone

identified findings

(1,2)

to

the

conversion

progesterone,

metabolites

that

pregnenolone

as intermediates,

the major

demonstrate

convert 8

pregnenolone

gesterone

moset

experiments

anubis

androstenedione

results

these

in

and the

17-hydroxypro-

incubates.

which we reported

in which

the

and

delta-4

The se

in

the

mar-

pathway

was

pre-

dominant. is

It

perhaps

significant

that

marily

through

terone

and 17-hydro~progest.erone

testosterone baboon

the delta-4

pathway,

testicular

fragments

the total

radioactivity

synthesis

within

within

delta-4

in fact

the delta-4

intermediates.

species,

the

the pathway for

a consequence pregnenolone genase

of

of

baboon

is metabolized

and isomerase

of

other

of

primate

pregnenolone

specific

primarily is

testosterone

by

and

3S-hydroxysteroid into

of

suggest

testosterone the

delta-5 than

are

non-primate

to testosterone

testicular

converted

tes-

portion

results

the intermediates

of

, pregnenolone

These

for

and

when

pathway,

substantial

into

to

17-hydroxypregnenolone

the delta-5 a

proges-

However,

predominant

converted

conversion

the activities

for

pri-

converted

testis.

with of

is

testis,

more readily

efficiently

incubates.

the

pathway

the baboon

pathway are

In the testis

accounted

occurred

intermediates

by the baboon

intermediates

and androstenedione

the

not

were

were incubated

tosterone

although

metabolism

the delta-4

or to androstenedione

and dehydroepiandrosterone,

that

although

enzymes.

is If

dehydro-

progesterone

and

a predominant is

metabolized

is

converted

delta-4 primarily into

delta-5

pathway

testis,

the

permit

the

(8,9).

of

pregnenolone

pathway.

pathway

are

and

more

to

a

baboon

lwer

with

conversion

of

by

baboon

predominant the

of

testosterone pathway

pregnenolone

may

3S-hydroxy-

consequent

intermediates

delta-4

the

17-hydroxylase

a

to

predominant in

progesterone

converted the

that

pregnenolone

the

testis,

pregnenolone

a consequent

suggest

to

although

efficiently

the

of

isomerase ,

Therefore,

pregnenolone

17-hydroxylase,

results

metabolism

if

However,

with

present activity

delta-4

due

The lower

by

(6,7).

pregnenolone

apparent

dehydrogenase

nant

by

results

17-hydroxypregnenolone

steroid

stenedione

pathway

de1 ta-5

and

may to

be

andropredomi-

17-hydroxypreg-

nenolone. We have

recently

Oedipus

where

mediates

of

the the

testosterone

it

subhuman

pathway

However,

through

appears

the

preferred

earlier

was

was

the

delta-5

there

by which

more

converted was

into

apparently

the

in

no

incubates.

As

differences

specie is

inter-

17-hydroxypregnenolone

since

substantial

Saguinus the

although

there

identified

pregnenolone

marmoset

efficiently

marmoset,

not

the

predominant

pathway

are

in

converted

within

testosterone

into

tissue.

results

from

substrate

efficiently

lone

in

results

were

were

that

primates

testicular The

was

delta-5

similar

pathway

dehydroepiandrosterone

such,

by

delta-4

(10).

conversion and

reported

these for

converted

poorly

converted

postulated

that

studies

baboon into

suggests

testicular

that

progesterone

into progesterone

17-hydroxypregnenolone. is

the

preferred

the

Progesterone

17-hydroxylase.

17_hydroxyprogesterone,

is

while

pregnenoTamaoki

substrate

(7) for

S 17-hydroxylase. reported

However,

that

TPEOXD-

Fevold

17-hydroxylase

(8,9)

and,

more

preferentially

recently,

converts

Kremers

(11) into

pregnenolone

17-hydroxypregnenolone. It

is

interesting

that

there

2&-dihydroprogesterone progesterone marmoset major

substrates. Callithrix

incubated

when

with reported

(12,131 gesterone

in

tosterone Brownie

the

predominant subhuman In

the

pathway.

delta-4

conversion

studies

metabolites

in

enzyme the

may

testis

Saguinus

to

of

progesterone Steinberger

a

the was

a

were nt

metabolite

fragments

which the

the

of from

biosynthesis

Rhesus

formation Sharma

monkey. of

aJ. pro-

normal,

of

al.

to

tes-

Hoschoian

testosterone

et --

pregnenolone

the

of

Papio

baboons,

incubates

increased

(1,2).

were

the by

step as

is

and occurs

reported

(5)

testosterone

in

a this

of

low

interesting

these and

we recently

the

testosterone reported

and

metabolites

within

2Oo-hydroxysteroid

conversion

C17-C201yase for

that

17-hydroxyprogesterone

17-hydroxylase

However,

rate-limiting anubis,

it

accumulation

androstenedione

be

Oedipus

in

However,

The

activities.

progesterone

humans, as

in

the

with

the

suggests

dehydrogenase

in

specie.

present

incubates

or

testicular

is

that

20a-dihydroprogesterone. the

In

primate

delta-5

primate

demonstrated

and

subjects.

investigated

reported

pregnenolone

pregnenolone

human

subhuman

been

(14)

through

of

of

2Oa-dihydroprogesterone

fragments.

Klinefelters

other

has

that

accumulation

both

previously

20a-dihydroprogesterone

and

only

(3)

significant with

have

radiolabelled

incubates

azoospermic, The

We

testicular

a

incubates

jacchus

metabolite

major

in

was

of

17-hydroxy-

suggests

that

synthesis in

the

this within

marmoset

Although we utilized only a three

hr

incubation

in

the

present

studies, which precluded the detection of an early conversion of nenolone to testosterone through the delta-5 pathway,

it

that any major conversion occurred through this pathway. 17-hydroxyprogesterone

and

20a-dihydroprogesterone

a

minor

Progesterone,

conversion and

epiandrosterone were identified in

in

incubates,

but

for

within

through the delta-5 pathway since 17-hydroxypregnenolone the

unlikely

accounted

substantial portion of the radioactivity from pregnenolone incubates. However, it is apparent that

is

preg-

a the

occurred dehydrovery

low

levels.

ACKNOWLEDGMENTS The skilled technical assistance of Mr. Charles E. Sutherland, Jr., is gratefully acknowledged. This work was supported by NICHD Grant P50-HD-08338. REFERENCES

1. Preslock, J.P., and E. Steinberger, STEROIDS 28, 775 (1976). 101 31 2. Preslock, J.P., and E. Steinberger, GEN COMP. ENDOCRINOL. -' (1977). 3. Preslock, J.P., and E. Steinberger, BIOL. REPROD. 17, 289 (1977). 547 33 4. Preslock, J.P., and E. Steinberger, GEN. COMP. EmINOL. _' (1977). 499 5. Sharma, D.C., S.G. Joshi, and R.I. Dorfman, ENDOCRINOLOGY 80, (1967). 6. Tamaoki, B.I., Il. Inano, and H. Nakano, In: The Gonads: McKerns, NHPC, 547 (1963). 7. Tamaoki, B.I., J. STEROID BIOCHEM. 4, 89 (1973). 8. Fevold, A.R., SCIENCE. 156 1753 (1767). 211 BIOCHEM. BIOPHYS. ACTA. -’313 9. Fevold, A.R., and H.B. &ond, (1973). 163 10. Preslock, J.P., and E. Steinberger, J. STEROID BIOCHEM. 2, (1978). 11. Kremers, P., J. STEROID BIOCHEM. 7, 571 (1976). 12. Steinberger, E., M. Ficher, and K.D. Smith, In: The Human Testis: Rosenberg and Paulsen (1970). 13. Steinberger, E., K.D. Smith, R.K. Tcholakian, M. Chowdhury, A. Steinberger, M. Ficher, and C.A. Paulsen, In: Male Sterility and Fertility. Mancini and Martini (1973). 14. Hoschoian, J.C., and A.C. Brownie, STEROIDS -10, 49 (1967).

S The following

trivial

201

TP&OXDO names were utilized

in the text:

Pregnenolone: 3g-hydroxy-5-pregnen-20-one Progesterone: 4-pregnene-3, 20-dione 17-Hydroxypregnenolone: 38, 17-dihydroxy-5-pregnen-20-one 17-Hydroxyprogesterone: 17-hydroxy-4-pregnene-3, 20-dione 20a-Dihydroprogesterone: 20a-hydroxy-4-pregnen-3-one Dehydroepiandrosterone: 3B-hydroxy-5-androsten-17-one Androstenedione: 4-androstene-3, 17-dione Testosterone: 17B-hydroxy-4-androsten-3-one 5a-androstanediol: 5a-androstane-3a, 17g-diol