Downregulation of AR may occur by aromatization of Test to Estrogen

[Dr.X]

Ok, the experimental systems employed aren't ideal for our purposes so keep that in mind, but nevertheless provide evidence for AR downregulation...by estrogen! Low levels of T seem to slightly increase AR expression but high levels (which are more likely to convert to E) inhibit the AR.

In regards to the second abstract, it is interesting that the use of proscar increases conversion to E (as expected) therefore may not be advisable without concommittant use of an anti-E.

Autoregulation of estrogen and androgen receptor mRNAs and downregulation of androgen receptor mRNA by estrogen in primary cultures of lizard testis cells.
Cardone A, Angelini F, Varriale B.
Gen Comp Endocrinol. 1998 Jun;110(3):227-36.

Steroid hormones regulate many developmental and physiological processes via specific receptors whose number can be up- or downregulated. The regulation of estrogen (ER) and androgen (AR) receptor mRNAs in primary cultures of lizard testis is described. The high degree of homology between the probes used and the receptor mRNAs in lizard testis was consistent with the high-stringency hybridisation conditions and the molecular size of ER mRNAs (7.4 and 4.5 kb) and AR mRNA (9.5 kb). Primary cultures of testis cells revealed a time- and drug-dependent relationship between ER and AR mRNAs. 17beta-oestradiol (E) autoregulated ER mRNA and downregulated AR mRNA. The antiestrogen ICI 164,384 reversed the latter effect. Cycloheximide (Cy), to inhibit protein synthesis, in combination with E, impaired the AR mRNA expression. Testosterone (T) autoregulated the expression of its own receptor mRNA whereas this effect was reversed by both flutamide (F) and Cy. Dose-response experiments showed that low concentrations of steroids (E or T 10(-12) M) increased ER or AR mRNA levels, respectively. These results suggest that both estrogen and androgen may autoregulate the expression of their own receptor mRNAs. Since in lizard testis androgens are significantly involved in meiosis and spermiogenesis and E dramatically impairs the AR mRNA expression, the latter effect may be key in regulating certain phases of reproduction.

Testosterone down-regulates the levels of androgen receptor mRNA in smooth muscle cells from the rat corpora cavernosa via aromatization to estrogens.
Lin MC, Rajfer J, Swerdloff RS, Gonzalez-Cadavid NF.
J Steroid Biochem Mol Biol. 1993 May;45(5):333-43.

Androgens down-regulate the levels of androgen receptors (AR) and AR mRNA in the penis and prostate of castrated rats, and are assumed to cause their decrease during sexual maturation in the penile smooth muscle of intact rats. In order to determine whether these effects occur directly at the target cell level, and to what extent they are due to testosterone (T) or to their metabolites, we have measured AR mRNA in cultures of smooth muscle cells from the adult rat corpora cavernosa treated in vitro with sex steroids. T at high concentrations (100 nM) acted like dihydrotestosterone (DHT) in increasing moderately the levels of AR mRNA in both proliferating and contact-inhibited cells. However, when conversion of T to DHT was blocked by the 5-alpha reductase inhibitor finasteride, the levels of AR mRNA were considerably down-regulated by T (10-500 nM), particularly in the contact-inhibited cells. Finasteride by itself was inactive. These effects in both types of cultures were inhibited by platelet derived growth factor (PDGF) (20 ng/ml), a growth factor that up-regulates AR mRNA levels, and by fadrozole (100 nM), an aromatase inhibitor of the T/estrogen conversion. Estradiol (50 nM) was even more potent than T in decreasing AR mRNA levels. With the exception of PDGF none of the treatments affected significantly cell growth, as measured by DNA synthesis and content. Our results indicate that it is possible to modulate in vitro AR mRNA levels in the penile smooth muscle cells, and that under normal conditions DHT and T act as moderate up-regulators. When DHT formation is inhibited, the aromatization pathway of T to estradiol will prevail and induce a pronounced down-regulation of AR mRNA levels. We assume that the in vivo AR down-regulation in the penile smooth muscle by androgens is an indirect effect mediated by a paracrine or endocrine mechanism elicited in another tissue.

 

[Ulter]

I am not a lizard or a rat. These studies do not translate to humans.

Taking issue with the idea of androgen receptor down-regulation.
By Bryan Haycock MS.

There is as much misinformation about steroids as there is good information had among bodybuilding enthusiasts. Go to any gym and you will hear some kid spouting off to his buddies about how steroids do this, or how they do that, or whatever. This soon starts somewhat of a pissing contest (excuse the expression) as to who knows more about steroids. It's the same kind of titillating and infectious banter that adolescent boys get into about girls and sex. With steroid banter you hear all the popular terms like Deca, Test, GH, gyno, zits, raisins, "h-u-u-u-ge", roid, freak, monster, roid-rage, "I knew this guy once", etc., etc.. If by some rare chance they are smart and have been reading this or some other high quality bodybuilding site on the net, they may actually get a few details right. More often than not they know just enough to be dangerous. Fortunately steroids haven't proven to be all that dangerous. Not only that, but most of these guys who are infatuated with steroids won't ever use or even see them except in magazines.

This kind of ego driven gym talk doesn't really bother me until they begin giving advice to other clueless people who actually have access to them. Spewing out steroid lingo gives other less experienced kids the impression that these kids actually know what they are talking about. That's how all of the psuedo-science folklore about steroids perpetuates. This is also why most people who actually use steroids know little about them. This last fact should bother anyone who cares about bodybuilding and/or bodybuilders.

I started out with this article planning on giving some textbook style explanation as to why using steroids doesn't down regulate androgen receptors (AR). Then after considering some of my critics views that I tend to write articles that hardly anyone can read, I decided to write an easy to read, yet informative explanation about what androgens actually do and how this precludes androgen receptor down regulation. I still have a few references but not so many that it looks like a review paper.

Androgen receptors down-regulate….Don't they?
One misunderstood principle of steroid physiology is the concept of androgen receptors (AR), sometimes called "steroid receptors", and the effects of steroid use on their regulation. It is commonly believed that taking androgens for extended periods of time will lead to what is called AR "down regulation". The premise for this argument is; when using steroids during an extended cycle, you eventually stop growing even though the dose has not decreased. This belief has persisted despite the fact that there is no scientific evidence to date that shows that increased levels of androgens down regulates the androgen receptor in muscle tissue.

The argument for AR down-regulation sounds pretty straightforward on the surface. After all, we know that receptor down-regulation happens with other messenger-mediated systems in the body such as adrenergic receptors. It has been shown that when taking a beta agonist such as Clenbuterol, the number of beta-receptors on target cells begins to decrease. (This is due to a decrease in the half-life of receptor proteins without a decrease in the rate that the cell is making new receptors.) This leads to a decrease in the potency of a given dose. Subsequently, with fewer receptors you get a smaller, or diminished, physiological response. This is a natural way for your body to maintain equilibrium in the face of an unusually high level of beta-agonism.

In reality this example using Clenbuterol is not an appropriate one. Androgen receptors and adrenergic receptors are quite different. Nevertheless, this is the argument for androgen receptor down-regulation and the reasoning behind it. The differences in the regulation of ARs and adrenergic receptors in part show the error in the view that AR down-regulate when you take steroids. Where adrenergic receptor half-life is decreased in most target cells with increased catecholamines, AR receptors half-live's are actually increased in many tissues in the presence of androgens.1

Let me present a different argument against AR down-regulation in muscle tissue. I feel that once you consider all of the effects of testosterone on muscle cells you come to realize that when you eventually stop growing (or grow more slowly) it is not because there is a reduction in the number of androgen receptors.

Testosterone: A multifaceted anabolic
Consider the question, "How do anabolic steroids produce muscle growth?" If you were to ask the average bodybuilding enthusiast I think you would hear, "steroids increase protein synthesis." This is true, however there is more to it than simple increases in protein synthesis. In fact, the answer to the question of how steroids work must include virtually every mechanism involved in skeletal muscle hypertrophy. These mechanisms include:

Enhanced protein synthesis

Enhanced protein synthesis

Enhanced growth factor activity (e.g. GH, IGF-1, etc.)

Enhanced activation of myogenic stem cells (i.e. satellite cells)

Enhanced myonuclear number (to maintain nuclear to cytoplasmic ratio)

New myofiber formation

Starting with enhanced growth factor activity, we know that testosterone increases GH and IGF-1 levels. In a study by Fryburg the effects of testosterone and stanozolol were compared for their effects on stimulating GH release.2 Testosterone enanthate (only 3 mg per kg per week) increased GH levels by 22% and IGF-1 levels by 21% whereas oral stanozolol (0.1mg per kg per day) had no effect whatsoever on GH or IGF-1 levels. This study was only 2-3 weeks long, and although stanozolol did not effect GH or IGF-1 levels, it had a similar effect on urinary nitrogen levels.

What does this difference in the effects of testosterone and stanozolol mean? It means that stanozolol may increase protein synthesis by binding to AR receptors in existing myonuclei, however, because it does not increase growth factor levels it is much less effective at activating satellite cells and therefore may not increase satellite cell activity nor myonuclear number directly when compared to testosterone esters. I will explain the importance of increasing myonuclear number in a moment, first lets look at how increases in GH and IGF-1 subsequent to testosterone use effects satellite cells.

Don't forget Satellite cells!
Satellite cells are myogenic stem cells, or pre-muscle cells, that serve to assist regeneration of adult skeletal muscle. Following proliferation (reproduction) and subsequent differentiation (to become a specific type of cell), satellite cells will fuse with one another or with the adjacent damaged muscle fiber, thereby increasing the number of myonuclei for fiber growth and repair. Proliferation of satellite cells is necessary in order to meet the needs of thousands of muscle cells all potentially requiring additional nuclei. Differentiation is necessary in order for the new nucleus to behave as a nucleus of muscle origin. The number of myonuclei directly determines the capacity of a muscle cell to manufacture proteins, including androgen receptors.

In order to better understand what is physically happening between satellite cells and muscle cells, try to picture 2 oil droplets floating on water. The two droplets represent a muscle cell and a satellite cell. Because the lipid bilayer of cells are hydrophobic just like common oil droplets, when brought into proximity to one another in an aqueous environment, they will come into contact for a moment and then fuse together to form one larger oil droplet. Now whatever was dissolved within one droplet (i.e. nuclei) will then mix with the contents of the other droplet. This is a simplified model of how satellite cells donate nuclei, and thus protein-synthesizing capacity, to existing muscle cells.

Enhanced activation of satellite cells by testosterone requires IGF-1. Those androgens that aromatize are effective at not only increasing IGF-1 levels but also the sensitivity of satellite cells to growth factors.3 This action has no direct effect on protein synthesis, but it does lead to a greater capacity for protein synthesis by increasing fusion of satellite cells to existing fibers. This increases the number of myonuclei and therefore the capacity of the cell to produce proteins. That is why large bodybuilders will benefit significantly more from high levels of androgens compared to a relatively new user.

Testosterone would be much less effective if it were not able to increase myonucleation. There is finite limit placed on the cytoplasmic/nuclear ratio, or the size of a muscle cell in relation to the number of nuclei it contains.4 Whenever a muscle grows in response to training there is a coordinated increase in the number of myonuclei and the increase in fiber cross sectional area (CSA). When satellite cells are prohibited from donating viable nuclei, overloaded muscle will not grow.5,6 Clearly, satellite cell activity is a required step, or prerequisite, in compensatory muscle hypertrophy, for without it, a muscle simply cannot significantly increase total protein content or CSA.

More myonuclei mean more receptors
So it is not only true that testosterone increases protein synthesis by activating genetic expression, it also increases the capacity of the muscle to grow in the future by leading to the accumulation of myonuclei which are required for protein synthesis. There is good reason to believe that testosterone in high enough doses may even encourage new fiber formation. To quote the authors of a recent study on the effects of steroids on muscle cells:

"Intake of anabolic steroids and strength-training induce an increase in muscle size by both hypertrophy and the formation of new muscle fibers. We propose that activation of satellite cells is a key process and is enhanced by the steroid use."7

Simply stated, supraphysiological levels of testosterone give rise to increased numbers of myonuclei and thereby an increase in the number of total androgen receptors per muscle fiber. Keep in mind that I am referring to testosterone and testosterone esters. Not the neutered designer androgens that people take to avoid side effects. This is not an argument to rapidly increase the dosages you use. It takes time for these changes to occur and the benefits of higher testosterone levels will not be immediately realized.

Maintenance of the kind of muscle mass seen in top-level bodybuilders today requires a given level of androgens in the body. That level will vary from individual to individual depending on their genetics. Nevertheless, if the androgen level drops, or if they were to "cycle off" the absolute level of lean mass will also drop. Likewise, as the level of androgens goes up, so will the level of lean mass that individual will be able to maintain. All of this happens without any evidence of AR down regulation. More accurately it demonstrates a relationship between the amount of androgens in the blood stream and the amount of lean mass that you can maintain. This does not mean that all you need is massive doses to get huge. Recruitment of satellite cells and increased myonucleation requires consistent "effective" training, massive amounts of food, and most importantly, time. Start out with reasonable doses. Then, as you get bigger you can adjust your doses upwards.

References:

Kemppainen JA, Lane MV, Sar M, Wilson EM. Androgen receptor phosphorylation, turnover, nuclear transport, and transcriptional activation. Specificity for steroids and antihormones. J Biol Chem 1992 Jan 15;267(2):968-74

Fryburg DA., Weltman A., Jahn LA., et al: Short-term modulation of the androgen milieu alters pulsatile, but not exercise- or growth hormone releasing hormone-stimulated GH secretion in healthy men: Impact of gonadal steroid and GH secretory changes on metabolic outcomes. J Clin Endocrinol. Metab. 82(11):3710-37-19, 1997

Thompson SH., Boxhorn LK., Kong W., and Allen RE. Trenbolone alters the responsiveness of skeletal muscle satellite cells to fibroblast growth factor and insulin-like growth factor-I. Endocrinology. 124:2110-2117, 1989

Rosenblatt JD, Yong D, Parry DJ., Satellite cell activity is required for hypertrophy of overloaded adult rat muscle. Muscle Nerve 17:608-613, 1994

Rosenblatt JD, Parry DJ., Gamma irradiation prevents compensatory hypertrophy of overloaded extensor digitorum longus muscle. J. Appl. Physiol. 73:2538-2543, 1992

Phelan JN, Gonyea WJ. Effect of radiation on satellite cell activity and protein expression in overloaded mammalian skeletal muscle. Anat. Rec. 247:179-188, 1997

Kadi F, Eriksson A, Holmner S, Thornell LE. Effects of anabolic steroids on the muscle cells of strength-trained athletes. Med Sci Sports Exerc 1999 Nov;31(11):1528-34

 

[Dr.X]

Ulter,
1. Most of the references used in Mr. Haycocks transcript were not from human studies either. In fact, due to the nature of anabolic investigations, there are hardly any human studies - therefore the dismissal of animal models in terms of applicability to human physiology is somewhat foolish. Yea, you have to consider there are probably differences but you can't ignore them.

2. The article doesn't provide any evidence that AR downregulation doesn't occur.

3. There is no solid evidence that new fiber formation occurs in humans, at least to a significant degree, and as such there will not be a corresponding increase in AR number.

Changes in testosterone muscle receptors: effects of an androgen treatment on physically trained rats.
Bricout VA, Germain PS, Serrurier BD, Guezennec CY.
IMASSA-CERMA, Departement de Physiologie Systemique, Bretigny sur Orge, France.

Cell Mol Biol (Noisy-le-grand). 1994 May;40(3):291-4.
From results obtained in physiological investigations carried out on various tissues sensitive to androgens, it seems that the hormonal receptivity can reflect changes in the endocrine status and specific response of a tissue. The purpose of the present investigation was to test whether an androgen treatment could modify the receptivity to testosterone of the skeletal muscle and myocardium of endurance trained rats. The experiment extended over 8 weeks, and animals received injections of delayed testosterone heptylate every seven days. The myocardium and two skeletal muscles with opposed functions and typology were examined: the extensorum digitorum longus (EDL), and the soleus (SOL). Results obtained using techniques based upon the radio-competition principles provided information on the testosterone-receptor binding. The binding curves were plotted up to the saturating concentration of tritiated mibolerone, a synthetic androgen specific of androgen receptors. The quantity of receptors, calculated at the specific saturation plateau is expressed in fmol/mg protein. Results show that contractile muscular activity always increased the quantity of receptors whereas the steroid treatment decreased it. Thus for EDL and SOL of control trained rats the quantity of receptors was 0.78 and 0.82 fmol/mg protein, respectively, compared to 0.23 and 0.43 fmol/mg protein for sedentary testosterone-treated rats. The same "contractile activity" effect was observed on the myocardium but enhanced with values of 1.63 fmol/mg protein for control trained rats versus 0.30 fmol/mg protein for sedentary testosterone-treated rats. The receptivity to testosterone of the skeletal muscle and myocardium changes under the effect of an androgen treatment.

 

[Ulter]

Right. Without any human study none of this means anything. If rat studies were all that was needed we wouldn't test humans would we? But humans and rats are not the same animal and so these studies on rats can be used to argue this both ways and neither makes any difference to us humans.
Like I said before, men who get testosterone replacement do not have to raise their doses which is anecdotal evidence that the AR doesn't downregulate. There are no studies showing otherwise in humans.

 

[Lumberg]

Ulter,
1. Most of the references used in Mr. Haycocks transcript were not from human studies either. In fact, due to the nature of anabolic investigations, there are hardly any human studies - therefore the dismissal of animal models in terms of applicability to human physiology is somewhat foolish. Yea, you have to consider there are probably differences but you can't ignore them.

2. The article doesn't provide any evidence that AR downregulation doesn't occur.

3. There is no solid evidence that new fiber formation occurs in humans, at least to a significant degree, and as such there will not be a corresponding increase in AR number.

Changes in testosterone muscle receptors: effects of an androgen treatment on physically trained rats.
Bricout VA, Germain PS, Serrurier BD, Guezennec CY.
IMASSA-CERMA, Departement de Physiologie Systemique, Bretigny sur Orge, France.

Cell Mol Biol (Noisy-le-grand). 1994 May;40(3):291-4.
From results obtained in physiological investigations carried out on various tissues sensitive to androgens, it seems that the hormonal receptivity can reflect changes in the endocrine status and specific response of a tissue. The purpose of the present investigation was to test whether an androgen treatment could modify the receptivity to testosterone of the skeletal muscle and myocardium of endurance trained rats. The experiment extended over 8 weeks, and animals received injections of delayed testosterone heptylate every seven days. The myocardium and two skeletal muscles with opposed functions and typology were examined: the extensorum digitorum longus (EDL), and the soleus (SOL). Results obtained using techniques based upon the radio-competition principles provided information on the testosterone-receptor binding. The binding curves were plotted up to the saturating concentration of tritiated mibolerone, a synthetic androgen specific of androgen receptors. The quantity of receptors, calculated at the specific saturation plateau is expressed in fmol/mg protein. Results show that contractile muscular activity always increased the quantity of receptors whereas the steroid treatment decreased it. Thus for EDL and SOL of control trained rats the quantity of receptors was 0.78 and 0.82 fmol/mg protein, respectively, compared to 0.23 and 0.43 fmol/mg protein for sedentary testosterone-treated rats. The same "contractile activity" effect was observed on the myocardium but enhanced with values of 1.63 fmol/mg protein for control trained rats versus 0.30 fmol/mg protein for sedentary testosterone-treated rats. The receptivity to testosterone of the skeletal muscle and myocardium changes under the effect of an androgen treatment.

OK so we are looking at two competing factors for regulation of AS receptors:

Exercise, which UPregulates, and

AS use (with no exercise), which downregulates.

Unfortunately, this study does not list the baseline androgen receptivity.

I am inclined to think that the exercise effect outweighs the AS effect. I bet you that if the study had taken baseline saturation figures they would have been closer to the sedentary AS treated rats than the exercising AS treated rats.

One knock against the study is that in AS usage in humans, the subject has usually trained for a significant period of time and has thus probably experienced some UPregulation of AS receptors due to what gaisn he has already made.

This is another strong argument for maximizing your genetic potential before starting AS usage. The bigger you already are, the bigger bang for your buck you are going to get on your first cycle.

JC