scholarly journals Androgen profiles during pubertal Leydig cell development in mice

Reproduction ◽  
2010 ◽  
Vol 140 (1) ◽  
pp. 113-121 ◽  
Author(s):  
Xiufeng Wu ◽  
Ramamani Arumugam ◽  
Ningning Zhang ◽  
Mary M Lee
Keyword(s):  
Leydig Cell ◽  
Developmental Changes ◽  
Side Chain ◽  
Adult Rat ◽  
Testosterone Production ◽  
Chain Cleavage ◽  
Cleavage Enzyme ◽  
Species Specific

Postnatal Leydig cell (LC) development in mice has been assumed empirically to resemble that of rats, which have characteristic hormonal profiles at well-defined maturational stages. To characterize the changes in LC function and gene expression in mice, we examined reproductive hormone expression from birth to 180 days, and quantified in vivo and in vitro production of androgens during sexual maturation. Although the overall plasma androgen and LH profiles from birth through puberty were comparable to that of rats, the timing of developmental changes in androgen production and steroidogenic capacity of isolated LCs differed. In mice, onset of androgen biosynthetic capacity, distinguished by an acute rise in androstenedione and testosterone production and an increased expression of the steroidogenic enzymes, cytochrome P450 cholesterol side-chain cleavage enzyme and 17α-hydroxylase, occurred at day 24 (d24) rather than at d21 as reported in rats. Moreover, in contrast to persistently high testosterone production by pubertal and adult rat LCs, testosterone production was maximal at d45 in mice, and then declined in mature LCs. The murine LCs also respond more robustly to LH stimulation, with a greater increment in LH-stimulated testosterone production. Collectively, these data suggest that the mouse LC lineage has a delayed onset, and that it has an accelerated pace of maturation compared with the rat LC lineage. Across comparable maturational stages, LCs exhibit species-specific developmental changes in enzyme expression and capacity for androgen production. Our results demonstrate distinct differences in LC differentiation between mice and rats, and provide informative data for assessing reproductive phenotypes of recombinant mouse models.

scholarly journals Evaluation of relative roles of LH and FSH in regulation of differentiation of Leydig cells using an ethane 1,2-dimethylsulfonate-treated adult rat model

2003 ◽  
Vol 176 (1) ◽  
pp. 151-161 ◽  
Author(s):  
V Sriraman ◽  
MR Sairam ◽  
AJ Rao
Keyword(s):  
Leydig Cells ◽  
Serum Testosterone ◽  
Side Chain ◽  
Mrna Levels ◽  
Rt Pcr ◽  
Lh Receptor ◽  
Side Chain Cleavage ◽  
Chain Cleavage ◽  
Cleavage Enzyme

The relative role of LH and FSH in regulation of differentiation of Leydig cells was assessed using an ethane 1,2-dimethylsulfonate (EDS)-treated rat model in which endogenous LH or FSH was neutralized from day 3 to day 22 following EDS treatment. Serum testosterone and the in vitro response of the purified Leydig cells to human chorionic gonadotropin (hCG) was monitored. In addition RNA was isolated from the Leydig cells to monitor the steady-state mRNA levels by RT-PCR for 17alpha-hydroxylase, side chain cleavage enzyme, steroidogenic acute regulatory protein (StAR), LH receptor, estrogen receptor (ER-alpha) and cyclophilin (internal control). Serum testosterone was undetected and the isolated Leydig cells secreted negligible amount of testosterone on stimulation with hCG in the group of rats that were treated with LH antiserum following EDS treatment. RT-PCR analysis revealed the absence of message for cholesterol side chain cleavage enzyme and 17alpha-hydroxylase although ER-alpha and LH receptor mRNA could be detected, indicating the presence of undifferentiated precursor Leydig cells. In contrast, the effects following deprival of endogenous FSH were not as drastic as seen following LH neutralization. Deprival of endogenous FSH in EDS-treated rats led to a significant decrease in serum testosterone and in vitro response to hCG by the Leydig cells. Also, there was a significant decrease in the steady-state mRNA levels of 17alpha-hydroxylase, cholesterol side chain cleavage enzyme, LH receptor and StAR as assessed by a semiquantitative RT-PCR. These results establish that while LH is obligatory for the functional differentiation of Leydig cells, repopulation of precursor Leydig cells is independent of LH, and also unequivocally establish an important role for FSH in regulation of Leydig cell function.

scholarly journals The compartmentation of non-esterified and esterified cholesterol in the superovulated rat ovary

1971 ◽  
Vol 123 (2) ◽  
pp. 143-152 ◽  
Author(s):  
A. P. F. Flint ◽  
D. T. Armstrong
Keyword(s):  
Microsomal Fraction ◽  
Specific Radioactivity ◽  
Subcellular Fractions ◽  
Side Chain ◽  
Electron Microscopic ◽  
Side Chain Cleavage ◽  
Chain Cleavage ◽  
Cleavage Enzyme ◽  
Rat Ovary

1. The specific radioactivities of non-esterified and esterified cholesterol, progesterone and 20α-hydroxypregn-4-en-3-one were determined in slices of superovulated rat ovary after incubation with [1-14C]acetate in vitro for various times. The specific radioactivities of progesterone and 20α-hydroxypregn-4-en-3-one were equal, and (during the fourth hour of incubation) exceeded those of the non-esterified cholesterol and the esterified cholesterol by factors of 2.8 and 7.6 respectively. 2. After separation of homogenates of superovulated rat ovary slices previously incubated with [14C]acetate into subcellular fractions by differential centrifugation, the specific radioactivities of non-esterified cholesterol in the cytosol, mitochondria, lipid-containing storage granules and microsomal fraction were 1220, 1510, 1420 and 4020d.p.m./μmol respectively; the corresponding values for the specific radioactivity of the esterified cholesterol were 600, 700, 730 and 760d.p.m./μmol. The specific radioactivities of progesterone and 20α-hydroxypregn-4-en-3-one were equal in all fractions; the corresponding mean specific radioactivity of progesterone+20α-hydroxypregn-4-en-3-one was 6150d.p.m./μmol. 3. By using glutamate dehydrogenase and cytochrome (a+a3) as mitochondrial markers, the presence of cholesterol side-chain cleavage enzyme was demonstrated in microsomal fraction free of mitochondrial contamination. 4. The specific radioactivities of ovarian non-esterified and esterified cholesterol, progesterone and 20α-hydroxypregn-4-en-3-one were determined up to 8h after the intravenous injection of [4-14C]cholesterol into superovulated rats. At all times the specific radioactivities of progesterone and 20α-hydroxypregn-4-en-3-one were equal to the specific radioactivity of non-esterified cholesterol and exceeded, by up to 3.3-fold, that of the esterified cholesterol. 5. It is concluded that non-esterified cholesterol formed from [14C]acetate in the endoplasmic reticulum equilibrates slowly with non-esterified cholesterol in other subcellular fractions, and is preferentially converted into steroids. Such a mechanism presupposes the operation of a microsomal cholesterol side-chain cleavage enzyme using non-esterified cholesterol as its substrate. Unrelated evidence is presented in support of the existence of such an enzyme. The results are discussed in the light of other biochemical and electron-microscopic findings relating to the compartmentation of cholesterol in steroidogenic tissues.

scholarly journals Activities of enzymes responsible for steroid biosynthesis and cholesterol ester metabolism in rabbit ovarian interstitial tissue and corpora lutea. A comparison of enzyme activities with flow rates

1973 ◽  
Vol 132 (2) ◽  
pp. 301-311 ◽  
Author(s):  
A. P. F. Flint ◽  
D. T. Armstrong
Keyword(s):  
Cholesterol Ester ◽  
Cholesterol Esterase ◽  
Interstitial Tissue ◽  
Corpora Lutea ◽  
Chain Cleavage ◽  
Cleavage Enzyme ◽  
Storage Granules ◽  
Ester Synthetase

A method involving the use of isolated cholesterol ester-storage granules as substrate is described for the assay of cholesterol esterase in rabbit ovarian tissues. Activities of cholesterol esterase 100–200-fold higher than those previously reported in ovarian tissues were measured by using this method. In addition to that of cholesterol esterase, activities of cholesterol ester synthetase, cholesterol side-chain cleavage enzyme and 3β-hydroxy steroid dehydrogenase were determined in rabbit ovarian interstitial tissue and corpora lutea. Activities of these enzymes are in general compatible with the flows through them measured under a variety of conditions both in vivo and in vitro. It is concluded that, in the rabbit ovarian tissues investigated, these enzymes are capable of catalysing the conversions usually attributed to them.

Transcriptional Regulation of Steroidogenic Genes: STARD1, CYP11A1 and HSD3B

2009 ◽  
Vol 234 (8) ◽  
pp. 880-907 ◽  
Author(s):  
Holly A. LaVoie ◽  
Steven R. King
Keyword(s):  
Transcriptional Regulation ◽  
Current Knowledge ◽  
Regulatory Protein ◽  
Side Chain ◽  
Combinatorial Regulation ◽  
Cytochrome P450scc ◽  
Chain Cleavage ◽  
Cleavage Enzyme ◽  
Steroidogenic Acute Regulatory ◽  
Species Specific

Expression of the genes that mediate the first steps in steroidogenesis, the steroidogenic acute regulatory protein (STARD1), the cholesterol side-chain cleavage enzyme, cytochrome P450scc (CYP11A1) and 3β-hydroxysteroid dehydrogenase/Δ5-Δ4 isomerase (HSD3B), is tightly controlled by a battery of transcription factors in the adrenal cortex, the gonads and the placenta. These genes generally respond to the same hormones that stimulate steroid production through common pathways such as cAMP signaling and common actions on their promoters by proteins such as NR5A and GATA family members. However, there are distinct temporal, tissue and species-specific differences in expression between the genes that are defined by combinatorial regulation and unique promoter elements. This review will provide an overview of the hormonal and transcriptional regulation of the STARD1, CYP11A1 and specific steroidogenic HSD3B genes in the adrenal, testis, ovary and placenta and discuss the current knowledge regarding the key transcriptional factors involved.

Hachimijiogan Produces Testosterone in Adult Rat Testes

1988 ◽  
Vol 16 (03n04) ◽  
pp. 93-105 ◽  
Author(s):  
Satoshi Usuki
Keyword(s):  
Male Rats ◽  
In Vivo Study ◽  
Steroid Production ◽  
Adult Rat ◽  
Testosterone Production ◽  
Incubation Study ◽  
Estradiol 17Β

The effect of Hachimijiogan (HZ) and Keishibukuryogan (KB) on the steroid production in rats was examined in vivo and in vitro. In an in vivo study, HZ stimulated the testes from ten-week old male rats to produce testoterone, whereas KB decreased the tissue testosterone concentrations. The Δ4-androstenedione and estradiol-17β (E2) showed no significant changes. In an incubation study, HZ also stimulated the testosterone production. The data suggested that HZ produces testosterone in rat testes. The role of KB is questionable.

Localization and functional activity of cytochrome P450 side chain cleavage enzyme (CYP11A1) in the adult rat kidney

2011 ◽  
Vol 332 (1-2) ◽  
pp. 253-260 ◽  
Author(s):  
Melina A. Pagotto ◽  
María L. Roldán ◽  
Romina M. Pagotto ◽  
María C. Lugano ◽  
Gerardo B. Pisani ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Functional Activity ◽  
Rat Kidney ◽  
Side Chain ◽  
Side Chain Cleavage ◽  
Adult Rat ◽  
Chain Cleavage ◽  
Cleavage Enzyme

SITES OF METYRAPONE INHIBITION OF STEROID BIOSYNTHESIS BY RAT ADRENAL MITOCHONDRIA

1974 ◽  
Vol 76 (4) ◽  
pp. 703-711 ◽  
Author(s):  
Andres Carballeira ◽  
Su Chiau Cheng ◽  
Lawrence M. Fishman
Keyword(s):  
Inhibitory Effect ◽  
Side Chain ◽  
Steroid Biosynthesis ◽  
Side Chain Cleavage ◽  
Chain Cleavage ◽  
Low Concentrations ◽  
Wide Range ◽  
Generating System

ABSTRACT Rat adrenal mitochondrial preparations supplemented with an NADPH-generating system were incubated with various labelled substrates in order to evaluate further the action of metyrapone on the utilization of cholesterol for steroid biosynthesis1). The formation of pregnenolone from [4-14C] cholesterol (0.5, 1.0 and 2.0 μCi) in 5, 10 and 15 min incubations was decreased by 72–82 % in the presence of metyrapone (0.5 mm). Similarly, the generation of labelled side chain fragments from [26-14C]-cholesterol was depressed 36–42 % by 0.2 mm metyrapone and 65–70 % by 1.0 mm inhibitor during 30, 60 and 90 min incubations. Metyrapone inhibition of the side chain cleavage was not observed, however, if cholesterol was replaced as substrate by its C-20 hydroxylated analog: The formation of pregnenolone from [7-3H]20α-hydroxycholesterol (0.5, 1.0 and 2.0 μCi), also an NADPH-mediated mitochondrial reaction, was not affected by similar concentrations of metyrapone, indicating that the inhibition observed with cholesterol as substrate is not related to non-specific toxic effects, to interference with NADPH generation or to impairment of NADPH function in the mitochondrial electron transport system. Parallel incubations with [4-14C] 11-deoxycorticosterone and with [4-14C] cholesterol over a wide range of inhibitor concentrations (0.01–1.0 mm) demonstrated that the effects of metyrapone on 11β-hydroxylation and on the side chain cleavage were dose-related; at low concentrations, however, metyrapone was a more potent inhibitor of 11β-hydroxylation than of cholesterol conversion to pregnenolone. These studies demonstrate clearly in the rat adrenal the dual inhibitory effect of metyrapone sug-gested by previous in vivo and in vitro observations in man.

Stimulatory and inhibitory factors of Leydig cell steroidogenesis are secreted simultaneously by the rat seminiferous tubules and do not affect Leydig cell inhibin production in vitro

1994 ◽  
Vol 6 (6) ◽  
pp. 693 ◽  
Author(s):  
JR McFarlane ◽  
Kretser DM de ◽  
GP Risbridger
Keyword(s):  
Conditioned Medium ◽  
Leydig Cell ◽  
Leydig Cells ◽  
Seminiferous Tubules ◽  
Low Doses ◽  
Adult Rat ◽  
Testosterone Production ◽  
Significant Reversal ◽  
The Mean

The effect of conditioned medium from rat seminiferous tubules (at Stages VII-VIII and Stages IX-VI) cultured with or without follicle-stimulating hormone (FSH) on the production of testosterone and immunoactive inhibin by Leydig cells was examined. Low doses of conditioned medium from unstimulated tubules at Stages VII-VIII significantly (P < 0.05) increased the mean testosterone production to greater than 31 +/- 11% over that achieved with luteinizing hormone (LH) alone. At the highest dose, the conditioned medium significantly inhibited (P < 0.05) LH-stimulated testosterone production by 13 +/- 7%. Low doses of conditioned medium from unstimulated tubules at Stages IX-VI increased the mean testosterone production to 22 +/- 10%, whereas at higher doses, a significant reversal in the stimulation occurred although not to the same extent as that found with medium from tubules at Stages VII-VIII. Conditioned medium from FSH-stimulated tubules at Stages VII-VIII and Stages IX-VI, significantly increased testosterone production to 39 +/- 7% and 31 +/- 13% respectively. Immunoactive inhibin production by the Leydig cells remained unaffected by exposure to conditioned medium from FSH stimulated and unstimulated tubules at Stages VII-VIII and Stages IX-VI. The data demonstrate that tubule culture medium contains FSH-modulated activities which can specifically stimulate and inhibit testosterone synthesis by adult rat Leydig cells in vitro and therefore explains the contradictory reports in the literature.

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