Lizard Liver Research Articles

Absorption, distribution, metabolism, and elimination of epoxiconazole enantiomers in lizards (Eremias argus)

Epoxiconazole (EPX) is a world widely used chiral triazole fungicide in the agriculture field. The excessive application of this triazole may cause damage to lizards. However, limited information is known about the toxicokinetics of EPX on lizards. Our study aimed to investigate the enantioselective absorption, distribution, metabolism, and elimination (ADME) of EPX in lizards following low and high dose exposure (10 and 100 mg kg−1 bodyweitht (bw)). The results demonstrated that (+)-EPX was easier absorbed than (−)-EPX in lizard plasma. Both (+)-EPX and (−)-EPX were detected in the liver, gonad, kidney, skin, brain, and intestine, with (+)-EPX preferentially distributed in these tissues. The elimination of (−)-EPX was faster than that of (+)-EPX in lizard liver and kidney in the high dose groups. Chiral conversion was found between EPX enantiomers in lizard skin. Simultaneously, five metabolites including M2, M4, M10, M18 and M19 were detected in lizard liver and kidney after EPX enantiomers exposure. The relative concentrations of M2, M4, and M10 were higher in the liver and kidney of (−)-EPX groups than those produced from (+)-EPX groups. The metabolic enzymes CYP3A4 and SULT1A1 primarily mediated enantioselective metabolism of EPX. The conclusions drawn from this study significantly enhance our understanding of the enantioselective behaviors of chiral triazole fungicides in reptiles, offering essential guidance for assessing the risks associated with different enantiomers of triazole fungicides.

Temperature-Dependent Bioaccumulation, Metabolism, and Hepatotoxicity of Flufiprole in Lizards (Eremias argus).

As a phenylpyrazole insecticide, flufiprole is an important substitute for fipronil in the agricultural field of China. However, its bioaccumulation and metabolism in terrestrial organisms especially in the lizards living in the agricultural area have rarely been investigated. As an ectothermic animal, lizards are also sensitive to temperature changes. Considering global warming, this study measured bioaccumulation, metabolism, and hepatotoxicity of flufiprole in the Chinese native lizard (Eremias argus) under different temperature stresses. Lizards exposed to flufiprole-contaminated soil adsorbed flufiprole through the skin and flufiprole was preferred to accumulate in lizard liver and brain. The oxidation product fipronil sulfone was the main metabolite of flufiprole in both lizard liver and human liver microsomes, which were mainly metabolized by lizard CYP3A19 or human CYP3A4. The fipronil sulfone concentration increased with increased temperature in lizard tissues. In addition, more serious oxidative damage was shown under higher temperature as the glutathione (GSH), malondialdehyde (MDA), and 8-hydroxy-2'-deoxyguanosine (8-OHdG) levels in lizards increased with increased temperature after flufiprole exposure. Flufiprole exposure also induced lizard liver lesions, and these lesions became more serious in the higher-temperature groups. This study provided new insights into the risk assessment of flufiprole in lizards under global warming.

Acute temperature adaptation mechanisms in the native reptile species Eremias argus

Reptiles are sensitive to temperature changes as ectotherm animals. The climate warming may pose more serious threat to reptiles. Although the behavior effect and reproduction biology have been well studied, little information is available about the adaptation mechanisms of reptiles to temperature stress. In this study, the native Chinese species, Eremias argus were incubated at 15 (cold stress), 25 (control group) and 35 °C (thermal stress) for 24 h. The transcriptome and metabolome technology were applied to investigate the molecular regulation mechanisms of lizards to acute temperature changes. The CIRBP and HSPA8 were hub genes in response to temperature adaptation. The increased expression of PER gene in lizard circadian rhythm is associated with tyrosine metabolism after cold or thermal stress. The poly-unsaturated fatty acids in female lizard liver were significantly increased with up-regulation of FASN and ACACA genes after thermal stress, which proved the disruption of fatty acid biosynthesis pathway in corporation with the altered body weight. The cortisol and testosterone were important steroid hormones in response to temperature changes especially in male lizard liver. The increased CIRBP gene expression in lizard gonads suppressed the KDM6B gene, which regulates the testis development and may induce sex reversal in male lizard after thermal stress. The adaptation responses of lizards to temperature stress may threaten the health status of wild population.

Lambda-cyhalothrin and its common metabolite differentially modulate thyroid disruption effects in Chinese lizards (Eremias argus)

Although the thyroid effects of pyrethroids on non-target organisms have been well studied, research on the toxic effects of pyrethorid metabolites is still limited. In this study, a type of representative Chinese lizards was used as the model and exposed to environmentally relevant concentrations of lambda-cyhalothrin (LCT) and 3-phenoxybenzoic acid (PBA) through cultivation on 3 and 15 μg/g soil to evaluate and compare their disruption effects on lizard hypothalamus-pituitary-thyroid (HPT) axis. The alterations occurred in lizards were examined through histopathology analyses, hormone level and gene expression measurements, the molecular binding interactions were analyzed in silico as well. The results showed that LCT exposure increased the plasma triiodothyronine (T3), thyroxine (T4) levels and the follicular epithelium heights of thyroid glands, whereas PBA induced no or much less degree of alterations. The ugt and dio2 gene expression in lizard liver was significantly up-regulated by LCT, but PBA caused less or opposite effects. The in silico homology simulation illustrated that LCT binds to TRα in the similar way of T3, while PBA binds to TRβ in the same manner of T3. The results demonstrated that both LCT and its metabolite-PBA could disrupt lizard HPT axis but through distinct mechanisms. The information would facilitate the comprehensive environmental safety assessment of pyrethroids.

Enantioselective accumulation, elimination and metabolism of fenbuconazole in lizards (Eremias argus)

The enantioselective accumulation, elimination and metabolism of fenbuconazole in lizards were determined following a single-dose (25 mg/kgbw) exposure to racemic or enantiomeric fenbuconazole. Accumulation of fenbuconazole was found in lizard fat with rac-form > enantiopure enantiomers. The enantiomer fractions (EFs) were higher than 0.5 in the blood, while EFs were less than 0.5 in the liver, brain, skin and stomach. There was conversion from (+)-fenbuconazole to (−)-fenbuconazole in lizard liver and conversion from (−)-fenbuconazole to (+)-fenbuconazole in lizard liver and blood. The results showed that enantioselective accumulation appeared in lizards, but the direction varied among blood and different tissues. The elimination half-lives (t1/2) of (+)-fenbuconazole were higher than those of (−)-fenbuconazole in the blood and liver, suggesting that (−)-fenbuconazole eliminated faster than (+)-fenbuconazole in these tissues. In addition, both (+)-fenbuconazole and (−)-fenbuconazole eliminated faster in the liver and stomach exposed to racemate than those exposed to enantiopure enantiomers. On the contrary, the form of racemate decreased the elimination rate of fenbuconazole in lizard fat. Synergistic elimination may occur when two enantiomers coexisted in lizard liver and stomach, while the racemate produced antagonistic elimination in lizard fat. Simultaneously, three metabolites, RH-6467, RH-9029&RH-9030 and keto-mchlorophenol, were discovered in lizard liver. Only two metabolites, RH-6467 and RH-9029&RH-9030, were found in lizard blood. RH-9029&RH-9030 were the major metabolites. The discovered enantiomers of (+)-fenbuconazole metabolites were different from those of (−)-fenbuconazole. The findings of this study may provide a better understanding of the enantioselective behaviors of chiral triazole fungicides in reptiles.

Environmental relevant concentration of λ-cyhalothrin and 3-phenoxybenzoic acid caused endocrine-disrupting effects on male lizards (Eremias argus)

In the present study, the endocrine toxicity of LCT and PBA was investigated through exposure to Eremias argus for two weeks under environmental relevant concentration. RNA-sequencing identified 4442 and 4653 differentially expressed genes in lizard liver after LCT and PBA exposure. Four differentially expressed genes (hsd17β, ar, sult, ugt) related with hypothalamic-pituitary-gonadal axis were quantified by qPCR. The expression of genes associated with HPG axis in different tissues differed significantly. In LCT treatment group, ar, cyp17 and hsd3β genes involved in testosterone synthesis and transportation were significantly decreased in lizard testes, and the spermatogensis was inhibited in the testes, which indicated the anti-androgenic activity of LCT. After PBA exposure, the genes related with estradiol synthesis, transportation and metabolism, such as hsd17β, erα, ugt in lizard liver were important biomarkers and the significant decrease of estradiol level was highly correlated with hsd17β, erα, ugt gene expressions. The relative high binding affinity of PBA with ERα further demonstrated the anti-estrogenic activity of PBA. Our results elucidate the different toxic mechanism of LCT and PBA on lizard endocrine system at environmental relevant concentration. Pyrethroids metabolism may cause more seriously toxicity rather than detoxification.

Myclobutanil accumulation, transcriptional alteration, and tissue injury in lizards (Eremias argus) treated with myclobutanil enantiomers

Enantioselective toxicokinetics, accumulation, and toxicity of myclobutanil were investigated by oral exposure of myclobutanil enantiomers to lizards. After a single oral administration, the absorption half-lives (t1/2Ka) and elimination half-lives (t1/2k) were in the range of 0.133–14.828 and 3.641–17.682 h, respectively. The absorption and elimination half-lives of (+)-myclobutanil showed no significant differences from those of (-)-myclobutanil in lizard blood, whereas preferential enrichment of (-)-enantiomer was observed in the liver, fat, skin, intestine, lung and kidney. In the bioaccumulation experiments, the residue of (-)-myclobutanil was detected in most tissues at 7, 14, and 28 days, while (+)-myclobutanil was found only in lizard skin, at a concentration lower than that of (-)-myclobutanil. Thus, (-)-myclobutanil was preferentially accumulated in lizards. The transcriptional responses of metabolic enzyme genes indicated that cytochrome P450 1a1 (cyp1a1), cyp2d3, cyp2d6, cyp3a4 and cyp3a7 played a crucial role in the metabolism of (+)-myclobutanil, whereas cyp1a1, cyp2d3, cyp2d6, cyp2c8, and cyp3a4 contributed to the metabolism of (-)-myclobutanil. The difference in metabolism pathways may be a reason for the enantioselectivity of myclobutanil in lizard. Myclobutanil also affected the expression of antioxidant enzyme genes, and the (+)-myclobutanil treatment might produce higher oxidative stress in lizard liver when compared with its antipode. Hepatic histopathological changes such as hepatocellular hypertrophy, nuclear pyknosis, vacuolation, and non-zonal macrovesicular lipid accumulation were observed in the liver of lizards for both (+)-myclobutanil and (-)-myclobutanil treatments. Thus, myclobutanil could affect lizard liver upon multiple exposure. The findings of this study provide specific insights into the enantioselective metabolism and toxicity of chiral triazole fungicides in lizards.

Dose-dependent effects of flufenoxuron on thyroid system of mature female lizards (Eremias argus) and their offspring

More and more studies are focusing on toxic effect of pesticides on lizards. However, the responses of different life-stage lizards to pesticides have not been reported. In this study, according to RNA-seq library data, thyroxine 5′-deiodinase activity showed significant difference between mature and immature lizard liver. In addition, triiodothyronine (T3) level in immature lizard serum was higher than that in mature lizard serum. Thus, we investigated the thyroid disruption of flufenoxuron with different concentrations (0, 5, 20 mg/kg) to both mature lizards and their offspring. No significant differences were observed in immature lizard body weight compared between control and exposure groups while the body weight of mature lizards was significantly decreased after flufenoxuron exposure. Moreover, the levels of thyroid hormones and the expression of thyroid related genes changed with exposure concentration of flufenoxuron and exhibited different regulation mechanism between mature and immature lizards. In immature lizard liver, trα, trβ, dio1 and ugt genes controlled the thyroid hormone levels interfered by flufenoxuron. In contrast, the thyroxine (T4) pathway in mature lizard liver was significantly disrupted especially by 20 mg/kg flufenoxuron exposure. This study elucidated the different thyroid disruption effects of flufenoxuron to lizards based on different exposure doses and lizard life stages.

Metabolism of Diflubenzuron in Lizard ( Eremias argus) and Comparative Toxicity of Diflubenzuron and Its Metabolite.

The metabolic process of diflubenzuron in rat or fish has been well studied, but little is known about its elimination pathway in lizard. The current study predicted the metabolic route of diflubenzuron in lizard feces and compared the toxicity of diflubenzuron and 4-chloroaniline on lizard thyroid system. The amido bond cleavage was the major route for diflubenzuron elimination in lizard feces. 4-Chloroaniline as the most toxic diflubenzuron metabolite was also abundant in feces. According to liver slices, 4-chloroaniline exposure induced significant changes of nuclear shape, while diflubenzuron exposure caused significant hepatocytes clustering. On the basis of thyroid hormone and thyroid-related gene levels, triiodothyronine (T3) level in lizard liver was regulated by thyroid hormone receptors, while thyroxine (T4) concentration was modulated by dio2 and udp genes after diflubenzuron or 4-chloroaniline exposure. These results showed that both diflubenzuron and 4-chloroaniline could disrupt lizard thyroid system, which could provide evidence for lizard population decline.

Stereoselective degradation and thyroid endocrine disruption of lambda-cyhalothrin in lizards (Eremias argus) following oral exposure

The disturbance of the thyroid system and elimination of chiral pyrethroid pesticides with respect to enantioselectivity in reptiles have so far received limited attention by research. In this study, bioaccumulation, thyroid gland lesions, thyroid hormone levels, and hypothalamus-pituitary-thyroid axis-related gene expression in male Eremias argus were investigated after three weeks oral administration of lambda-cyhalothrin (LCT) enantiomers. In the lizard liver, the concentration of LCT was negatively correlated with the metabolite-3-phenoxybenzoic acid (PBA) level during 21 days of exposure. (+)-LCT exposure induced a higher thyroid follicular epithelium height than (−)-LCT exposure. The thyroxine levels were increased in both treated groups while only (+)-LCT exposure induced a significant change in the triiodothyronine (T3) level. In addition, the expressions of hypothalamus-pituitary-thyroid axis-related genes including thyroid hormone receptors (trs), deiodinases (dios), uridinediphosphate glucuronosyltransferase (udp), and sulfotransferase (sult) were up-regulated after exposure to the two enantiomers. (+)-LCT treatment resulted in higher expression of trs and (−)-LCT exposure led to greater stimulation of dios in the liver, which indicated PBA-induced antagonism on thyroid hormone receptors and LCT-induced disruption of thyroxine (T4) deiodination. The results suggest the (−)-LCT exposure causes higher residual level in lizard liver while induces less disruption on lizard thyroid activity than (+)-LCT.

Caveolae and caveolin-1 in reptilian liver

Caveolae are plasma-membrane invaginations that, by interacting with membrane-associated molecules such as endothelial nitric oxide synthase and tyrosine kinases, precisely regulate cell-signalling pathways responsible for cell structure and cell function. Indeed, there is widespread evidence that caveolae associate, structurally and functionally, with proteins, lipids and solutes to facilitate transcellular transport of these macromolecules. Caveolin-1, one of the family of membrane proteins that form caveolae, is most prominently expressed in endothelial cells of the vascular bed. Therefore, we have applied advanced electron microscopy as well as molecular biology techniques to study the presence of caveolae and caveolin-1 in the liver sinusoidal endothelium of reptiles. Reptiles are known to store excess lipid in the liver as an energy source for hibernation, and so offer a useful animal model in which to assess the structural and functional implications these subcellular compartments might have on liver sinusoidal endothelial transport. This study demonstrates that caveolae are indeed conserved across vertebrate species, whether mammalian or reptilian. It also presents as first novel data on the presence of caveolin-1-associated, tubular structures located within the cytoplasm of the lizard liver sinusoidal endothelium.

Influence of Temperature on the Liver Circadian Clock in the Ruin Lizard Podarcis sicula

Reptiles represent an interesting animal model to investigate the influence of temperature on molecular circadian clocks. The ruin lizard Podarcis sicula lives in a continental climate and it is subjected to wide range of environmental temperatures during the course of the year. As consequence, ruin lizard daily activity pattern includes either the hibernation or periods of inactivity determined by hypothermia. Here we showed the rhythmic expression of two clock genes, lPer2 and lClock, in the liver of active lizards exposed to summer photo-thermoperiodic conditions. Interestingly, the exposition of lizards to hypothermic conditions, typical of winter season, induced a strong dampening of clock genes mRNA rhythmicity with a coincident decrease of levels. We also examined the qualitative and quantitative distribution of lPER2 and lCLOCK protein in different cellular compartments during the 24-h cycle. In the liver of active lizards both proteins showed a rhythmic expression profile in all cellular compartments. After 3 days at 6 degrees C, some temporal fluctuations of the lCLOCK and lPER2 are still detectable, although, with some marked modifications in respect to the values detected in the liver of active lizards. Besides demonstrating the influence of low temperature on the lizard liver circadian oscillators, present results could provide new essential information for comparative studies on the influence of temperature on the circadian system across vertebrate classes.

Purification and Characterization of Liver Lectins from a Lizard, Sceloporus spinosus

This study discusses the purification of soluble β‐galactose lectins obtained from the lizard liver of Sceloporus spinosus. The first lectin named lizard hepatic lectin‐1 (LHL‐1) presented a molecular weight of 31,750, with an isoelectric point of 4.25. The highest specific hemagglutinating activity was achieved using human blood type A1: N‐acetilgalactosamine (GalNAc)–galactose (Gal)–fucose (Fuc). Carbohydrate inhibition assays indicated a higher lectin specificity for GalNAc. For LHL‐2 the molecular weight obtained was 23,850 with an isoelectric point of 3.25. The highest carbohydrate specificity was observed for Gal. These lizard hepatic lectins are similar to the mammal hepatic lectins previously reported. However, it is different from the alligator hepatic lectin (AHL). The homology analyses of LHL‐1 resulted in 100% identity with the Steroidogenic acute regulatory protein (StAR), while LHL‐2 was similar to adenylate kinase (75% identity). We suggest that these liver lectins are related to the inherent functions of liver previously reported.

The Tooth Puller [L'arracheur de dents

FROM THE MIDDLE AGES through the early 19th century, tooth pulling was often performance art. Barber-surgeons cut hair, set bones, let blood, and pulled teeth. Toothache could be treated with leeches, blistering, cupping, and laxatives, as well as with prescriptions of lizard liver, green frogs, and a urine gargle.1 Opium mixtures were popular painkillers. When refined sugar became widely available in the 17th century, dental caries became common. For more desperate sufferers, itinerant tooth pullers set up shop in marketplaces and at fairs. The “dentist” and his assistants attracted a crowd by telling stories, singing and dancing, performing tricks, or juggling. The tooth puller's assistant was usually dressed as a clown or a harlequin, with a pointed hat on which was inscribed the insignia of Saint Apollonia, the patron saint of toothache sufferers. In more elaborate cases, such as the one depicted in this image from late 18th-century France, the tooth puller (arracheur de dents) had a stage as well as 2 assistants. The music that they provided attracted the crowd and also helped to drown out the cries of the patient. The tooth extraction was an essential part of the entertainment. The public setting provided considerable drama and let the gathered crowd judge the skill of the “dentist.” If he were smooth enough—if he could extract teeth without causing too much pain—others might volunteer as patients. Many artists tried to capture the pomp, pageantry, and pain of early dentistry. This etching was created by Jean Duplessis-Bertaux (1747–1819), a well-known artist and engraver who chronicled many scenes of the French Revolution and the Napoleonic era.

Hepatic Outer-Ring Deiodinase in a Mexican Endemic Lizard (Sceloporus grammicus)

The kinetic characterization of the outer-ring deiodination pathway using rT3 (rT3-ORD) in male, female, and pregnant female livers of an endemic lizard, Sceloporus grammicus, is reported. The ORD pathway does not have the characteristics of deiodinase type II; it is exclusively carried out by deiodinase type I (DI). DI enzymatic activity in lizard liver contains one of the highest activities reported in vertebrates. This activity is sexually dimorphic, with males presenting the highest activity during the reproductive season. The properties of this enzyme correspond to those described in mammals, such as specificity for rT3, susceptibility to inhibition by 6-n-propyl-2-thiouracil and gold–thioglucose, cofactor requirement, and kinetic pattern. Unlike other vertebrates, the lizard DI exhibits conspicuous stability in the thermal range of 15 to 42°C and in the pH range of 5.0 to 9.0. Male true kinetic constants exhibit a direct correlation with temperature. This is in agreement with short-term adaptation to microenvironmental changes and the feasible expression of enzymatic forms/variants which, together, endow this lizard species with a greater adaptation to natural daily ambient thermal fluctuations.

Regulation of skeletal muscle metabolism in the lizard Dipsosaurus dorsalis by fructose-2,6-bisphosphate.

Changes in liver and skeletal muscle fructose-2,6-bisphosphate (Fru-2,6-P2) concentrations were compared during fasting, exercise, and recovery in the lizard Dipsosaurus dorsalis and in outbred mice (Mus musculus). We present the first correlative evidence that suggests that a decrease in the content of Fru-2,6-P2 may mediate elevated gluconeogenesis in lizard skeletal muscle. Contents of Fru-2,6-P2 in lizard gastrocnemius and red and white iliofibularis (IF) were significantly lower (as much as 55% in white IF) during recovery from exhaustive exercise than at rest. Recovery from exhaustive exercise had no significant effect on Fru-2,6-P2 concentrations in any mouse muscle examined. Fasting significantly depressed lizard and mouse liver Fru-2,6-P2 contents and decreased lizard red IF by over 84% from the fed condition. Lizard red and white muscle fiber bundles incubated in 20 mM lactate had significantly lower Fru-2,6-P2 (94 and 61% depression, respectively) than those incubated in 8.5 mM glucose. These results are consistent with the hypothesis that Fru-2,6-P2 acts as a signal for controlling gluconeogenesis in lizard skeletal muscle.

Evolution of energy metabolism. Proton permeability of the inner membrane of liver mitochondria is greater in a mammal than in a reptile

Standard metabolic rate is 7-fold greater in the rat (a typical mammal) than in the bearded dragon, Amphibolurus vitticeps (a reptile with the same body mass and temperature). Rat hepatocytes respire 4-fold faster than do hepatocytes from the lizard. The inner membrane of isolated rat liver mitochondrial has a proton permeability that is 4-5-fold greater than the proton permeability of the lizard liver mitochondrial membrane per mg of mitochondrial protein. The greater permeability of rat mitochondria is not caused by differences in the surface area of the mitochondrial inner membrane, but differences in the fatty acid composition of the mitochondrial phospholipids may be involved in the permeability differences. Greater proton permeability of the mitochondrial inner membrane may contribute to the greater standard metabolic rate of mammals.

Fructose-2,6-bisphosphate and other metabolites and enzymes in the process of cold-induced lethargy and starvation in lizard liver

1. 1. The levels of the metabolites fructose-2,6-bisphosphate, glycogen, ATP and ADP have been determined in the lizard liver along the process of cold induced lethargy and starvation. 2. 2. The levels of the key glycolytic enzymes glucokinase plus hexokinase, phosphofructokinase, pyruvate kinase, and lactic dehydrogenase have been also measured along the process. With the exception of glucokinase plus hexokinase, all the enzymes behave in the same way in both treatments. On the other hand the enzyme phosphofructokinase is strongly affected by short term exposure to cold. 3. 3. The data reported here show a similar long term response pattern of liver glycolysis to the conditions of cold exposure and starvation. Moreover, the behavior of metabolites and enzymes is very similar to that observed in mammalian starvation, except for glycogen.

The vascular anatomy of the liver of the monitor lizard (Genus varanus).

AbstractThe gross and microscopic anatomy of the vasculature of the monitor lizard liver was studied. The portal vein has a peculiar arrangement of smooth muscle. The tunical media of the entering portal vein has bundles of smooth muscle cells separated by large numbers of collagenous fibers. The amount of smooth muscle decreases as the vessel decreases in diameter and soon one finds intermittent broad, thin bands of smooth muscle. As the caliber of the vessels continues to decrease, the smooth muscle bands become narrower and thicker so that they appear as doughnut‐shaped sphincters. The sphincters are usually found at the beginning of each branch of the portal vein as well as along the course of veins between areas of branching. Some sphincters are found in direct contact with the outer capsule of the liver. Sphincters occur in the terminal branches of the portal vein just proximal to the sinusoids.Small numbers of scattered smooth muscle cells were seen arranged longitudinally, obliquely, or circularly in the smaller hepatic veins. Even the large hepatic veins had only small amounts of smooth muscle. At no place along the course of hepatic veins could smooth muscle sphincters equivalent to those seen around portal veins be found.The monitor lizard should be an excellent subject for physiological and pharmacological studies of regulation of intrahepatic portal vein blood flow.

Ageing changes in the nucleic acid and protein content of the liver of the garden lizard, Calotes versicolor

The relative DNA content (mg/g wet wt.) of the lizard liver declined sharply during postnatal stage remaining almost constant during the laterhalf of the life-span. The total DNA (mg/whole liver wet wt.) was found to increase with increasing bodyweight. The relative RNA content (mg/g wet wt.) reached the peak level at 16 g body weight and declined thereafter. The total RNA (mg/whole liver wet wt.) showed the same tendency as observed for total DNA. The relative protein content (mg/g liver wet wt.) showed the highest concentration at about 30 g bodyweight and remaining constant thereafter. When expressed on the basis of DNA (mg protein/mg DNA) the protein content also reached a peak level at about 30 g body weight but showed decline thereafter. The total protein (mg/whole liver wet wt.) indicated a steady rise with increasing age but at much slower rate beyond 50 g body weight. The RNA/ DNA ratio was highest at about 16 g body weight and declined thereafter. The RNA/Protein ratio also showed the similar trend.