Change in the alkaline phosphatase activity and glycogen content in the leukocytes of white rat blood under the influence of penicillin preparations]

The contents of alkaline phosphatase and glycogen in the neutrophils of the peripheral blood of unbred albino rats weighing 170-200 gm were determined. Benzylpenicillin and oxacillin in doses of 50000 and 20000 Units/kg respectively were administered intramuscularly once a day over a long period of time. Two-phase changes in the contents of alkaline phosphatase and glycogen under the effect of penicillin were found. An increase in their contents was observed on the 5-10th days then it was followed by a decrease. Increase in the contents of the metabolites proceeded simultaneously with enhancing of the digestive capacity of the leucocytes and activity of lysozyme in them.     

Adaptive response of hypertrophied skeletal muscle to endurance training

The response of hypertrophied soleus and plantaris muscle of rats to endurance training was studied. Hypertrophy was produced by bilateral extirpation of the gastrocnemius muscle. A 13-wk training program of treadmill running initiated 30 days after removal of the gastrocnemius muscle accentuated (P less than 0.01) the hypertrophy. Succinate dehydrogenase activities of the enlarged muscles of sedentary rats were similar to those of normal animals, as were the increases associated with training. Phosphorylase and hexokinase activities were unaltered as a result of the experimental perturbations. Rates of glycogen depletion during exercise were lower (P less than 0.01) in the liver and soleus and plantaris muscles of endurance-trained animals. No difference existed in the rate of glycogen depletion of normal and hypertrophied muscle within the sedentary or trained groups. These data demonstrate that extensively hypertrophied muscle responds to training and exercise in a manner similar to that of normal muscle.     

Modulation of radiation-induced biochemical alterations in mice by rosemary (Rosemarinus officinalis) extract

Radioprotective effect of leaves extract of Rosemarinus officinalis (ROE) has been studied against 6 Gy gamma-radiations in the liver of Swiss albino mice at various post-irradiation intervals between 12 h and 30 days. In control animals (without ROE treated irradiated), an elevation in glycogen, protein, acid and alkaline contents was found till day 5th, but thereafter decreased at successive intervals without returning to normal. Cholesterol level was found to be lower than normal till 10th day, then increased up to 20th day but later declined without restoring normal level. A similar trend of variation in these biochemical parameters was observed in experimental group (ROE pretreated irradiated) also but to a lower extent. ROE significantly delayed and inhibited the rise in these biochemical parameters. Almost normal values of such constituents were regained by day 30th in experimental animals; whereas in control animals, normal values were not ever attained. In control animals, there was an elevation in lipid peroxidation (LPx) and a decrease in glutathione (GSH) in blood and liver; whereas in experimental group, decline in LPx accompanied by an increase in GSH concentration was observed.     

Training increases muscle blood flow in rats with peripheral arterial insufficiency

This study investigated the effect of physical training on muscle blood flow (BF) in rats with peripheral arterial insufficiency during treadmill running. Bilateral stenosis of the femoral artery of adult rats (300-350 g) was performed to reduce exercise hyperemia in the hindlimb but not limit resting muscle BF. Rats were divided into normal sedentary, acute stenosed (stenosed 3 days before the experiment), stenosed sedentary (limited to cage activity), and stenosed trained (run on a treadmill by a progressively intense program, up to 50-60 min/day, 5 days/wk for 6-8 wk). Hindlimb BF was determined with 85Sr- and 141Ce-labeled microspheres at a low (20 m/min) and high treadmill speed (30-40 m/min depending on ability). Maximal hindlimb BF was reduced to approximately 50% normal in the acute stenosed group. Total hindlimb BF (81 +/- 5 ml.min-1.100 g-1) did not change in stenosed sedentary animals with 6-8 wk of cage activity, but a redistribution of BF occurred within the hindlimb. Two factors contributed to a higher BF to the distal limb muscle of the trained animals. A redistribution BF within the hindlimb occurred in stenosed trained animals; distal limb BF increased to approximately 80% (P less than 0.001) of the proximal tissue. In addition, an increase in total hindlimb BF with training indicates that collateral BF has been enhanced (P less than 0.025). The associated increase in oxygen delivery to the relatively ischemic muscle probably contributed to the markedly improved exercise tolerance evident in the trained animals.     

Histochemical studies on the effect of thyroid hormone on alkaline and acid phosphatase activities in liver of fish and amphibia.

The Lata fishes (Ophicephalus punctatus) showed increased alkaline and acid phosphatase activities in liver after immersion for 15-30 days in thyroxine-containing medium (0.025 mug/ml). A single injection of thyroxine (1-2 mug/g of body weight) caused increased acid phosphatase activity in liver of Lata fish in comparison to the controls on the 5th day after experiment but the alkaline phosphatase activity remained unchanged. Both alkaline and acid phosphatases showed increased activities in liver of Lata fishes treated with a single injection of 4 mug of thyroxine per g of body weight on the 5th day. Immersion of Lata fishes in thiourea solution (1 mg/ml) for 15 days did not show any alteration in alkaline or acid phosphatase activities but these enzyme activities decreased after 30 days' immersion in thiourea solution in comparison to the controls. A seasonal variation of alkaline and acid phosphatase activities was observed in liver of Lata fishes. More alkaline phosphatase activity was found in liver of summer fishes than in winter fishes. The winter fishes showed more acid phosphatase activity than the summer fishes. Three consecutive injections of thyroxine (0.1 mug/g of body weight) to toads (Bufo melanostictus) caused increased alkaline and acid phosphatase activities in liver on the 5th day of the experiment, in comparison to the controls.     

Exercise and exhaustion effects on glycogen synthesis pathways

Gunderson, Hans, Nadja Wehmeyer, Diane Burnett, John Nauman,Cynthia Hartzell, and Scott Savage. Exercise and exhaustion effects on glycogen synthesis pathways. J. Appl.Physiol. 81(5): 2020-2026, 1996.FemaleSprague-Dawley rats were infused with [1-¹³C]glucose tomeasure the effect of endurance training and the effect of variousmetabolic conditions on pathways of hepatic glycogen synthesis. Fourmetabolic states [sedentary (S), trained (T), sedentary exhausted(SE), and trained exhausted (TE)] were studied. T and TE ratswere trained on a motor-driven treadmill (30 m/min, 15% grade, 1.0 h/day, 5 days/wk) for 8-10 wk. After a 24-h fast, SE and TE ratswere run to exhaustion (sedentary average = 78 min, trained average = 155 min) at a training pace and immediately infused with labeledglucose for 2 h. S and T rats were infused after a 24-h fast. Afterinfusion, tissues were removed and glycogen was isolated and hydrolyzedto glucose. The glucose was measured for distribution of¹³C by using nuclear magneticresonance. Glycogen was synthesized predominantly by the indirectpathway for all metabolic states, indicating that infused glucose wasfirst metabolized primarily in the peripheral tissue. Thedirect-pathway utilization was greater in rested S than in rested Tanimals (30 vs. 14%); however, for exhausted animals, the trained useof the direct pathway was greater (22 vs. 9%). Both TE and rested Tanimals utilize the indirect pathway a comparable amount. Sedentaryanimals, on the other hand, dramatically decreased utilization of thedirect pathway, with exhaustive exercise changing from 30 to 9%. Theresults indicate that endurance training modifies glucose utilizationduring glycogen synthesis after fasting and exhaustive exercise.

     

Effect of exhaustive ultra-endurance exercise in muscular glycogen and both Alpha1 and Alpha2 Ampk protein expression in trained rats

Glycogen is the main store of readily energy in skeletal muscle and plays a key role in muscle function, demonstrated by the inability to sustain prolonged high-intensity exercise upon depletion of these glycogen stores. With prolonged exercise, glycogen depletion occurs and 5′-AMP-activated protein kinase (AMPK), a potent regulator of muscle metabolism and gene expression, is activated promoting molecular signalling that increases glucose uptake by muscular skeletal cells. The aim of this study was primarily to determine the effect of ultra-endurance exercise on muscle glycogen reserves and secondly to verify the influence of this type of exercise on AMPK protein expression. Twenty-four male Wistar rats, 60 days old, were divided into four experimental groups: sedentary, sedentary exhausted (SE), endurance trained (T) and endurance trained exhausted (TE). The animals ran for 10 to 90 min/day, 5 days/week, for 12 weeks to attain trained status. Rats were killed immediately after the exhaustion protocol, which consisted of running on a treadmill (at approximately 60 % V _max until exhaustion). Optical density of periodic acid-Schiff was detected and glycogen depletion observed predominantly in type I muscle fibres of the TE group and in both type I and II muscle fibres in the SE group. Plasma glucose decreased only in the TE group. Hepatic glycogen was increased in T group and significantly depleted in TE group. AMPK protein expression was significantly elevated in TE and T groups. In conclusion, acute exhaustive ultra-endurance exercise promoted muscle glycogen depletion. It seems that total AMPK protein and gene expression is more influenced by status training.     

Exercise effects on chromium excretion of trained and untrained men consuming a constant diet

Chromium excretion of eight trained and five sedentary men was determined on rest days and after exercise to exhaustion at 90% of maximum O2 consumption (VO2max) to determine if degree of physical fitness affects urinary Cr losses. Subjects were fed a constant daily diet containing approximately 9 micrograms Cr/1,000 kcal. VO2max of the trained runners was in the good or above range based on their age and that of the sedentary subjects was average or below. While consuming the control diet, basal urinary Cr excretion of subjects who exercise regularly was significantly lower than that of the sedentary control subjects, 0.09 +/- 0.01 and 0.21 +/- 0.03 microgram/day (mean +/- SE), respectively. When subjects consumed self-chosen diets, basal urinary Cr excretion of the trained subjects was also significantly lower than that of the untrained subjects. Daily urinary Cr excretion of trained subjects was significantly higher on the day of a single exercise bout at 90% VO2max compared with nonexercise days, 0.12 +/- 0.02 and 0.09 +/- 0.01 microgram/day, respectively. Urinary Cr excretion of sedentary subjects was not altered after controlled exercise. These data demonstrate that basal urinary Cr excretion and excretion in response to exercise are related to VO2max and therefore degree of physical fitness.     

Sub-chronic diclofenac sodium induced alterations of alkaline phosphatase activity in serum and skeletal muscle of mice

The present study has been carried on changes in activity of alkaline phosphatase in serum and gastrocnemius muscle of mice after sub-chronic use of diclofenac. Mice in experimental group received diclofenac (10 mg/kg body wt /day) for 30 days while control group received normal saline. Alkaline phosphatase was assayed in muscle and serum and its activity was localized histochemically in muscle. Results showed that diclofenac induced changes in specific activity of alkaline phosphatase at different periods of treatment variably compared to control group. Specific activity of alkaline phosphatase decreased significantly in gastrocnemius initially (48.74%), increased thereafter (132.96%) and slight decrease (13.97%) was noticed after 30 days. In serum, the specific activity of alkaline phosphatase decreased slightly after 10 days (18.78%), increased in the middle of the treatment period (132.04%) as well as showed increase (109.09%) compared to control group after 30 days stage of investigation. These findings were also confirmed by electrophoretic studies in muscle.     

Water exercise prevents femur density loss associated with ovariectomy in the retired breeder rat

The effect of non-weight-bearing exercise on skeletal bone remains controversial. The objective of this pilot study was to examine the effects of water exercise training on femur density and serum alkaline phosphatase activity in ovariectomized and sham-operated (ovaries left intact) retired breeder rats. Exercised animals swam at progressively increasing duration from 5 minutes to 75 min.d(-1), 5 d.wk(-1), for a 6-week conditioning period. Exercised rats had greater (p < 0.02) soleus muscle citrate synthase activity than sedentary rats, confirming an aerobic training effect. Femur density (g.cm(-3)) was greater (p < 0.0007) for exercised rats than sedentary rats but lower (p < 0.01) for ovariectomized rats compared to sham rats. Serum alkaline phosphatase activity tended (p < 0.06) to be greater for exercised rats compared to sedentary rats. These results indicate that dynamic water-flotation exercise prevents the femur bone loss associated with ovariectomy in rats. We conclude that this form of exercise could be beneficial in maintaining bone density in hormone-deficient postmenopausal women, especially the elderly who may not be able to perform weight-bearing activities.     

The effects of swimming and running regimens on skeletal muscle glycogen in the rat.

Endurance capacity and the effects of different post-exercise states on skeletal muscle glycogen have been studied in rats trained by swimming or running and in sedentary controls. Regular endurance exercise resulted in increased skeletal muscle glycogen stores. A greater depletion was observed in trained animals than in non-trained animals after a training bout or exhaustive exercise. While muscle glycogen levels did not reflect a differential training stimulus (running vs swimming), swimming as a measure of exhaustive exercise was deemed invalid because of the ability of trained swimmers to avoid stenuous exercise by an alteration of swimming pattern.     

Post-exercise ketosis and the glycogen content of liver and muscle in rats on a high carbohydrate diet

Post-exercise ketosis is known to be suppressed by physical training and by a high carbohydrate diet. As a result it has often been presumed, but not proven, that the development of post-exercise ketosis is closely related to the glycogen content of the liver. We therefore studied the effect of 1 h of treadmill running on the blood 3-hydroxybutyrate and liver and muscle glycogen concentrations of carbohydrate-loaded trained (n = 72) and untrained rats (n = 72). Resting liver and muscle glycogen levels were 25%-30% higher in the trained than in the untrained animals. The resting 3-hydroxybutyrate concentrations of both groups of rats were very low: less than 0.08 mmol.l-1. Exercise did not significantly influence the blood 3-hydroxybutyrate concentrations of trained rats, but caused a marked post-exercise ketosis (1.40 +/- 0.40 mmol.l-1 h after exercise) in the untrained animals, the time-course of which was the approximate inverse of the changes in liver glycogen concentration. Interpreting the results in the light of similar data obtained after a normal and low carbohydrate diet it has been concluded that trained animals probably owe their relative resistance to post-exercise ketosis to their higher liver glycogen concentrations as well as to greater peripheral stores of mobilizable carbohydrate.     

Histochemical response of mice to mistletoe lectin I (ML I)

Summary The acute toxicity of lectin ML I from the toxic drug, mistletoe, was demonstrated in previous experiments. Because the reason for this extremely high toxicity is not yet clear, mice were studied histochemically at different times after treatment with various doses of ML I, ML I A or ML I B chain separately, or recombinations of ML I A and ML I B. Various plasma membrane-associated hydrolases as well as Golgi apparatus-and endoplasmic reticulum-linked hydrolases, peroxisomal and extraperoxisomal oxidases, lysosomal hydrolases, mitochondrial dehydrogenases, the cytoskeletal proteins keratin and vimentin as well as iron, glycogen and lipids were analysed in all organs and tissues of female mice. Irrespective of the dose, a clear-cut response was only observed in the liver. After ML I treatment, glycogen disappeared completely from all hepatocytes, and this effect did not depend on the ML I-concentration and exposure time. The increase in activity of Golgiassociated thiamine pyrophosphatase in hepatocytes and of non-specific alkaline phosphatase in the sinusoidal endothelial cells depended on the applied ML I concentration and the time of treatment. Doses of 600 or 900 ng ML I/kg drastically increased the phosphatase activities. These clear-cut changes of glycogen and enzyme activities were not observed after administration of the ML I B chain alone, and less so when the mice were treated only with the ML I A chain, or were treated with a recombination of ML I A and ML I B even at concentrations higher than that of ML I. The glycogen as well as thiamine pyrophosphatase and non-specific alkaline phosphatase responses after ML I administration were highly reproducible. None of the other investigated cell constituents responded to treatment with ML I, irrespective of the organs, tissues and cells analysed. This was also true for mice treated with ML I concentrations higher than the LD₅₀, after which the animals died with 2–3 days. In conclusion, application of ML I induces clear-cut liver cell changes, which are not observed after treatment with individual constituents of the ML I molecule either singly or in recombination. However, these changes do not explain the deaths of the animals.Supported by WTZ (Vereinbarung über wissenschaftlich-technische Zusammenarbeit: Projekt 13)     

Activities before and after stress of digestive enzymes in rats separated at different time from lactating females

The effect of separation of rat pups from lactating females at different time after birth (at the 18th, 21st, 30th, 36th or 43rd day) and stress exposed at the day of the experiment (at the 75th day of life) or twice (at the day of weaning and at the day of the experiment) on activities of membrane-bound (invertase, alkaline phosphatase, aminopeptidase M) and predominantly intracellular (glycyl-L-leucine dipeptidase) enzymes was studied in different parts of intestine of rats (males and females). A change of the enzyme activities has been shown for the first time in the animals separated from lactating females not only at early time, which has already been known, but also at later time, in comparison with their activities in the animals weaned at usual time, at the 30th day of life. The preliminary stress of rat pups at the day of separation from lactating females promotes in many cases a reduction of the response to the stress, to which the animal is exposed in the adult period of life. Deceased.     

Cytochemical studies of rat liver glycogen and alkaline phosphatase under the effect of hepatocarcinogens.

Cytochemical changes of glycogen and alkaline phosphatase were studied under sequential effect of p-dimethylaminoazobenzene (DAB) feeding and CCl4 injections. It has been found that in liver cells of experimental animals, the concentration of glycogen and the activity of alkaline phosphatase were decreased.     

Influence of diet on the storage, mobilization and utilization of energy reserves in trained and untrained rats

The effect of the major dietary energy source (fat or carbohydrate) on some of the adaptations to physical training, particularly body composition and tissue glycogen concentrations, were studied in growing male Wistar rats. Resting liver glycogen concentrations were lower in both trained and sedentary rats fed a high fat diet compared to corresponding rats fed a high carbohydrate (low fat) diet. Trained rats on both diets had higher liver glycogen levels than corresponding sedentary controls. Resting gastrocnemius muscle glycogen concentrations were not influenced by diet or training. Rates of liver and muscle glycogen depletion during a 60-min swim were lower in trained rats but were not influenced by diet. Significant interactions were noted between the dietary energy source and exercise training with respect to body weight gain, body fat content, liver weight and liver glycogen concentrations.     

Treatment of immature mice with gonadotrophins. The influence of mouse age on the response of ovarian alkaline phosphatase activities to gonadotrophins

Treatment of mice aged 23-25 days with chorionic gonadotrophin induced large amounts of an ovarian alkaline phosphatase activity (phosphatase Ib) kinetically distinct from that of untreated ovaries (phosphatase I). The activities of alkaline phosphatase I and Ib varied with age in untreated mice. Phosphatase Ib appeared when serum luteinizing hormone concentrations increased (days 4-10 and days 35-45), and disappeared when concentrations were low (days 11-35). Injection of human chorionic gonadotrophin induced progressively larger amounts of phosphatase Ib activity between day 19 and day 29. However, gonadotrophin treatment failed to induce this activity on days 10-18 and 30-35. Nevertheless, during the latter period, human chorionic gonadotrophin induced especially large increases in uterine weight. Treatment at different ages with sheep luteinizing hormone plus human pituitary follicle-stimulating hormone induced a pattern of response identical with that induced by human chorionic gonadotrophin, although sheep luteinizing hormone alone was ineffective before 35 days. In contrast, human luteinizing hormone induced a response in the absence of exogenous follicle-stimulating hormone.     

Investigations of alkaline phosphatase Ca+2-ATPase and Na+, K+-ATPase during beta-APN-induced initial bone mineralization inhibition

Tibias of 6-day-old white Leghorn chick embryos treated with beta-aminopropionitrile (beta-APN; 0.1 mg/egg/day) for 4 days and injected with 3H-proline or 3H-tetracycline on the 11th day were analyzed for incorporation of 3H-proline and 3H-tetracycline. The incorporation of 3H-proline was comparable in the controls and beta-APN-treated embryos. However, the incorporation of 3H-tetracycline was significantly lower in beta-APN-treated embryos. The bone ash contents were also lower in the latter group. Alkaline phosphatase and Ca+2-ATPase were found to be significantly lower in beta-APN-treated embryonic bones. There was, however, no difference in the activity of Na+, K+-ATPase. The histochemical examination showed the alkaline phosphatase to be present on osteoblasts and matrix vesicle plasma membranes at the periosteal surface. The chick embryonic liver tissue showed no significant differences in the activities of any of the above enzymes. The results suggest that beta-APN-induced inhibition of the bone mineralization may be due to the bone-specific inhibition of alkaline phosphatase and Ca+2-ATPase.     

Gene expression of TGF-beta, TGF-beta receptor, and extracellular matrix proteins during membranous bone healing in rats

Poorly healing mandibular fractures and osteotomies can be troublesome complications of craniomaxillofacial trauma and reconstructive surgery. Gene therapy may offer ways of enhancing bone formation by altering the expression of desired growth factors and extracellular matrix molecules. The elucidation of suitable candidate genes for therapeutic intervention necessitates investigation of the endogenously expressed patterns of growth factors during normal (i.e., successful) fracture repair. Transforming growth factor beta1 (TGF-beta1), its receptor (Tbeta-RII), and the extracellular matrix proteins osteocalcin and type I collagen are thought to be important in long-bone (endochondral) formation, fracture healing, and osteoblast proliferation. However, the spatial and temporal expression patterns of these molecules during membranous bone repair remain unknown. In this study, 24 adult rats underwent mandibular osteotomy with rigid external fixation. In addition, four identically treated rats that underwent sham operation (i.e., no osteotomy) were used as controls. Four experimental animals were then killed at each time point (3, 5, 7, 9, 23, and 37 days after the procedure) to examine gene expression of TGF-beta1 and Tbeta-RII, osteocalcin, and type I collagen. Northern blot analysis was used to compare gene expression of these molecules in experimental animals with that in control animals (i.e., nonosteotomized; n = 4). In addition, TGF-beta1 and T-RII proteins were immunolocalized in an additional group of nine animals killed on postoperative days 3, 7, and 37. The results of Northern blot analysis demonstrated a moderate increase (1.7 times) in TGF-beta1 expression 7 days postoperatively; TGF-beta1 expression returned thereafter to near baseline levels. Tbeta-RII mRNA expression was downregulated shortly after osteotomy but then increased, reaching a peak of 1.8 times the baseline level on postoperative day 9. Osteocalcin mRNA expression was dramatically downregulated shortly after osteotomy and remained low during the early phases of fracture repair. Osteocalcin expression trended slowly upward as healing continued, reaching peak expression by day 37 (1.7 times the control level). In contrast, collagen type IalphaI mRNA expression was acutely downregulated shortly after osteotomy, peaked on postoperative days 5, and then decreased at later time points. Histologic samples from animals killed 3 days after osteotomy demonstrated TGF-beta1 protein localized to inflammatory cells and extracellular matrix within the fracture gap, periosteum, and peripheral soft tissues. On postoperative day 7, TGF-beta1 staining was predominantly localized to the osteotomized bone edges, periosteum, surrounding soft tissues, and residual inflammatory cells. By postoperative day 37, complete bony healing was observed, and TGF-beta1 staining was localized to the newly formed bone matrix and areas of remodeling. On postoperative day 3, Tbeta-RII immunostaining localized to inflammatory cells within the fracture gap, periosteal cells, and surrounding soft tissues. By day 7, Tbeta-RII staining localized to osteoblasts of the fracture gap but was most intense within osteoblasts and mesenchymal cells of the osteotomized bone edges. On postoperative day 37, Tbeta-RII protein was seen in osteocytes, osteoblasts, and the newly formed periosteum in the remodeling bone. These observations agree with those of previous in vivo studies of endochondral bone formation, growth, and healing. In addition, these results implicate TGF-beta1 biological activity in the regulation of osteoblast migration, differentiation, and proliferation during mandibular fracture repair. Furthermore, comparison of these data with gene expression during mandibular distraction osteogenesis may provide useful insights into the treatment of poorly healing fractures because distraction osteogenesis has been shown to be effective in the management of these difficult clinical cases.     

Changes in osteoblast, chondrocyte, and adipocyte lineages mediate the bone anabolic actions of PTH and small molecule GSK-3 inhibitor

Parathyroid hormone (PTH) and glycogen synthase kinase-3 (GSK-3) inhibitor 603281-31-8, administered once daily increased bone formation in vivo. We investigated the molecular mechanisms of the anabolic responses of PTH and 603281-31-8 in rat osteopenia model. Female 6-month-old rats were ovariectomized (Ovx) and permitted to lose bone for 1 month, followed by treatment with PTH (1-38) at 10 microg/kg/day s.c. or 603281-31-8 at 3 mg/kg/day p.o. for 60 days. Twenty-four hours after the last treatment, RNA from distal femur metaphysis was subjected to gene expression analysis. Differentially expressed genes (P<0.05) were subjected to pathway analysis to delineate relevant bio-processes involved in skeletal biology. Genes involved in morphogenesis, cell growth/differentiation, and apoptosis were significantly altered by Ovx and the treatments. Analysis of morphogenesis genes showed an overrepresentation of genes involved in osteogenesis, chondrogenesis, and adipogenesis. A striking finding was that Ovx decreased several markers of osteogenesis/chondrogenesis and increased markers of adipogenesis/lipid metabolism. Treatment with either PTH or the GSK-3 inhibitor reversed these effects, albeit at different levels. Histological analysis confirmed that osteopenia in Ovx animals was associated with three-fold increase in marrow adiposity. PTH and GSK-3 inhibitor restored bone volume, and reversed or normalized marrow adiposity. Ex vivo studies showed that PTH and GSK-3 inhibitor increased the ratio of colony forming marrow stromal progenitors (CFU-fs) that were alkaline phosphatase positive (putative osteoblasts). Our results suggest that the bone anabolic actions of PTH and GSK-3 inhibitor in vivo involve concerted effects on mesenchymal lineages; osteoblasts, chondrocytes, and adipocytes.