Transgenerational marking of freshwater fishes with enriched stable isotopes: a tool for fisheries management and research

A promising new method of marking larval freshwater fishes with enriched stable isotopes by means of injecting the maternal parent with the marking agent was investigated. The ¹³⁸Ba:¹³⁷Ba ratios in the otoliths of larval golden perch Macquaria ambigua were compared to determine the effect of injecting female broodstock with different dosages of enriched ¹³⁷Ba at various times before spawning. There was 100% mark success in the progeny of fish injected with 20 μg g⁻¹ of enriched ¹³⁷Ba 24 h before inducing spawning with hormones and 40 μg g⁻¹ administered at the same time as inducement of spawning. Injection of 40 μg g⁻¹ enriched ¹³⁷Ba 21 days before spawning resulted in only 81% mark success and suggests rapid elimination of barium in M. ambigua . Injection with enriched ¹³⁷Ba did not significantly affect the fertilization rate, number of fertilized eggs or hatching rate compared with long-term hatchery records. These results suggest that transgenerational marking is an effective and affordable means of mass-marking larval fishes. Thousands of larval fishes can be permanently marked with a unique artificial isotopic mark via a single injection into the maternal parent, thus avoiding the handling of individual fishes or having to deal with chemical baths. Because no single mark or tagging method is suitable for all situations, transgenerational marking with enriched stable isotopes provides another method for researchers and managers to discriminate both hatchery-reared and wild fishes.     

Otolith resorption induced by anaerobic stress in the goldfish, Carassius auratus

To examine otolith resorption induced by anaerobic stress, ⁴⁵Ca-prelabelled goldfish, Carassius auratus , were kept in oxygen-deficient ambient water (O₂ < 0.5 .1 1⁻¹, 26° C) for 2 days and otoliths (asterisci) were analysed for ⁴⁵Ca retention. In a second experiment, fish were anaerobically stressed for 2 days, and then received a single intraperitoneal injection of ⁴⁵Ca. They were maintained under stress for one more day and killed to examine ⁴⁵Ca deposition in otoliths. Plasma was analysed for total and radioactive calcium. Otoliths (lapilli) were examined for stress-induced check formation by scanning electron microscopy (SEM).
Stress significantly reduced plasma calcium levels and the rate of calcium retention in otoliths, which was calculated from the ⁴⁵Ca specific activity of the plasma. On the other hand, the rate of calcium deposition in otoliths was the same in the stressed as in the control fish. SEM observation revealed that the applied stress resulted in a check formation in otoliths. These results indicate that a 48-h stress of oxygen deficiency induces calcium resorption in otoliths.     

Effects of egg composition and prey density on the larval growth and survival of lake whitefish (Coregonus clupeaformis Mitchill)

Eggs were collected from two stocks of lake whitefish, Coregonus clupeaformis , in Lakes Michigan and Huron to assess the effect of egg composition and prey density on larval growth and survival. Egg composition parameters including wet weight (mg egg⁻¹), dry weight (mg egg⁻¹), percent water, total caloric content (cal egg⁻¹), caloric density (cal egg⁻¹), percent lipid content, and total lipidcontent (mg egg⁻¹) were measured. Fish hatched from six parental females in each stock were fed one of four rations (0, 18, 24, 50 brine shrimp larva⁻¹ day⁻¹) after yolk sac absorption. Length at hatch, endogenous growth, exogenous growth, and survival were measured during a 42-day laboratory experiment. Length at hatch of larvae was positively related to egg caloric content ( r ²=0.780). Endogenous growth for lake whitefish larvae was positively related to percent lipid content ( r ²=0.896) and total egg lipid content ( r ²=0.876) of parental females. Exogenous growth and survival of larval lake whitefish was positively related to prey availability. Larval fish growth was accurately modelled ( r ²=0.973) as a function of prey abundance using a threshold-corrected hyperbolic equation. These results indicate that both egg composition and prey availability have the potential to influence the growth and survival dynamics of larval lake whitefish significantly.     

VITAMIN B6 TRANSPORT IN THE CENTRAL NERVOUS SYSTEM: IN VIVO STUDIES

Abstract— The total concentrations of vitamin B₆ (B₆) in plasma, choroid plexus, CSF and brain of adult New Zealand white rabbits, measured fluorometrically, were 0.30, 15.10, 0.39 and 8.90 μ mol/l or kg respectively. The mechanisms by which B₆ enters and leaves brain, choroid plexus and CSF were investigated by injecting [³H]pyridoxine (PIN) intravenously, intraventricularly and intraarterially. [³H]PIN, with or without unlabelled PIN, was infused intravenously at a constant rate into conscious rabbits. At 150 min, [³H]B₆ readily entered CSF, choroid plexus and brain. The addition of 0.5 mmol/kg carrier PIN to the infusion solution depressed the relative entry of [³H]B₆ into CSF, choroid plexus and brain by about 80%. After intraventricular injection, [³H]PIN readily entered brain from CSF. The intraventricular injection of carrier PIN with [³H]PIN decreased the amount of [³H]B₆ in brain and also decreased the percentage of [³H]B₆ in CSF and brain that was phosphorylated. During one pass through the cerebral circulation, [³H]PIN (1 μ m ) was cleared from the circulation no more rapidly than mannitol. These results were interpreted as showing that the entry of B₆ from blood into CSF and presumably the extracellular space of brain and thence into brain cells involves one or more saturable transport and/or metabolic steps.     

Sibling cannibalism among juvenile vundu under controlled conditions. II. Effect of body weight and environmental variables on the periodicity and intensity of type II cannibalism

Cannibalism among starved groups of juvenile (19–48 days old) vundu catfish Heterobranchus longifilis was 66·5% nocturnal, and its impact under modified day length was proportional to the duration of the dark phase. Shallow depth and high population density decreased the intensity of cannibalism, whereas low density and deeper environments had an opposite effect. The presence of refuges had no significant effect on cannibalism. The maintenance ( R _maint) and maximum ( R _max) daily food rations (% day⁻¹) of cannibals feeding on live prey were modelled as R _maint=3·899 W _C^0·327 ( r ²=0·684; d.f.=31), and R _max=49.545 W _C^0·321 ( r ²=0·999; d.f.=5), where W _C was the body weight of the cannibal (g). The latter model indicated that the impact of a cannibal on a population decreased by a 20% margin each time the cannibal doubled its body weight, and suggested that cannibalism among vundu would become insignificant for cannibals heavier than 30 g. The significance of these findings is discussed within the contexts of vundu aquaculture and of general, conceptual models of the dynamics of cannibalism among fishes.     

Can the chemistry of otolith nuclei determine population structure of Patagonian toothfish Dissostichus eleginoides?

Otolith nucleus chemistry resolved the population structure of Patagonian toothfish Dissostichus eleginoides , an exclusively marine species, along the Patagonian Shelf and North Scotia Ridge out to South Georgia in the Southern Ocean. Concentrations of ⁵⁵Mn, ⁸⁸Sr and ¹³⁷Ba, ratioed to ⁴²Ca, showed a sharp population boundary in the vicinity of the Polar Front, between South Georgia and the North Scotia Ridge. These results validated otolith nucleus chemistry as a technique for examining population structure in Patagonian toothfish, demonstrating that otolith nucleus chemistry can discriminate between populations even in fully marine environments. Moreover, the nucleus chemistry indicated population heterogeneity not previously detected, suggesting the possibility of more than one South American population, and also suggested some South American-caught fish had moved from South Georgia.     

Relationship between opercular girth, maximum girth and total length of fishes caught in gillnets in the estuarine and lower river sections of Shatt al-Arab River (Basrah Province, Iraq)

In fisheries, the two morphological parameters of opercular and maximum girth are related to the effectivity of capture methods in gilled and wedged fish, respectively. The present work investigates the relationship between opercular ( G _ope) and maximum girths ( G _max) to total length ( L _t) for 10 fish species captured from Shatt al-Arab River, Basrah, Iraq. Data were collected October 2005 to December 2006. Cyprinids were the best represented family with six species; engraulids, silurids, heteropneustids and mugilids were each represented by one species. G _ope and G _max were found to increase linearly with total length of all species, all r ² values being greater than 0.73 and statistically significant (P < 0.01). When G _ope and G _max for all species were plotted against total length, two groups were identified ( G 1, G 2), corresponding to general girth-length relationships: (a) G 1 = −0.252 + 0.424 L _t and G 2 = −0.262  +  0.600 L _t for opercular girth and, (b) G 1 = 1.538 + 0.419 L _t and G 2  =  1.538  +  0.696 L _t for maximum girth. These groups correspond to different body shapes of fishes: G 1-round and G 2-compressed. These relationships have implications when using length data and mesh size to determine size selectivity of gill nets.     

Toxic effects of mercury on testicular activity in the freshwater teleost, Clarias batrachus (L.)

Catfish ( Clarias batrachus L.) were exposed to selected sublethal concentrations of mercuric chloride (HgCl₂; 0.05 mg l⁻¹), methylmercuric chloride (CH₃HgCl; 0.04 mg l⁻¹) and emisan 6 (an organic mercurial fungicide; 0.5 mg l ⁻¹) for intervals of 45, 90 and 180 days from February to July (preparatory to spawning phase of the annual reproductive cycle). The gonadosomatic index showed a significant decrease after 90- and 180-day exposure to the mercurials. Histologically, the seminiferous tubules were smaller in size and contained mostly spermatids in comparison to the control fish in which they were greatly distended and full of spermatozoa. The Leydig cells showed pycnotic changes in Hg-treated fish after 90 and 180 days. In the Hg-treated testes, total lipid content and ³²P uptake decreased significantly after 90 and 180 days. Levels of phospholipids and free cholesterol registered a significant reduction during all the durations. Esterified cholesterol level showed a significant decrease only in the 90-day HgCl₂ and CH₃HgCl groups, and in all 180-day Hg groups, while the level of free fatty acid decreased significantly only in the 180-day Hg groups. The observations suggest that impairment of testicular lipid metabolism by Hg is one of the possible factors that led to the inhibition of steroidogenesis and spermatogenesis.     

The Binding of Cations to the Surfaces of Cells from Early Chick Blastoderms

Cells of the early chick blastoderms are either preparing for or undergoing regulated morphogenetic movements which culminate in the formation of a three-layered embryo. Information on the changes in the physical-chemical properties of cell surfaces may help in the understanding of this process. The binding of magnesium, manganese, strontium, barium and lanthanum to surfaces of early embryonic cells was estimated by the changes induced by these cations in the cells' electrophoretic mobilities (EPM). Cells show a positive EPM at concentrations of MgCl₂ and MnCl₂ at 3 × 10⁻² M while SrCl₂, and BaCl₂ were not able to reverse the cells' charge at concentrations up to 6 × 10⁻² M. CaCl₃ reversed the cells' EPM at concentrations as low as 5 × 10⁻³ M.
Our results suggest that the surfaces of early embryonic cells have a high affinity for Mg and Mn. This is indicated by a reversal of polarity which cannot be detected in cells of differentiating or adult tissues at the cation concentrations used in these experiments.     

Endocrine, osmoregulatory, respiratory and haematological parameters in flounder exposed to the water soluble fraction of crude oil

Flounders Pleuronectes flesus with an implanted vascular catheter were exposed to a 50% dilution of the water soluble fraction (WSF) of Omani crude oil (c. 6ppm total aromatic hydrocarbons) and serial blood samples taken to determine their endocrine status (cortisol, catecholamines and thyroid hormones) and the resultant and/or causal physiological (haematological, ionoregulatory and respiratory) disturbances. This resulted in a progressive increase in plasma cortisol concentrations from 3 h onwards (rising from 18 to 51 ng ml⁻¹ after 48-h exposure), and increased plasma glucose concentrations indicating a generalized stress response. Plasma T3 and T4 concentrations of both control and WSF-exposed groups declined progressively over the experimental period; exposure to the WSF of crude oil further depressed plasma T4 concentrations but not plasma T₃ concentrations relative to those of control fish. Plasma osmolality and sodium and chloride concentrations were stable in WSF-exposed fish, however, plasma potassium concentrations were increased significantly at the 24-and 48-h sampling points. The most profound physiological disturbance in WSF-exposed fish was a dramatic decline in blood oxygen content (CvO₂) (from 2–8 to 0–8 ml 100 ml⁻¹ after 48-h exposure), which is likely to be the cause of the increased plasma noradrenaline concentrations from 3 h onwards. Increased noradrenaline is likely in turn to have been responsible for the significant increase in blood haematocrit and blood haemoglobin at the 3-h sampling point, although the dominant effect in the longer-term was a significant decline in both of these haematological parameters.     

Effect of algal deposition on acetate and methane concentrations in the profundal sediment of a deep lake (Lake Constance)

Abstract In the profundal sediment ot Lake Constance (143 m depth) the temperature is constant at 4 °C. Despite the constant temperature, CH₄ concentrations changed with season between about 120 μM in winter and about 750 μM in summer, measured down to 30 cm depth. The acetate concentration profiles also varied between seasons. In summer, acetate concentration reached a maximum at about 100 μM in 2 or 4 cm depth. In winter, maximal concentrations of about 5 μM were observed over the entire depth. Input of organic material in late spring may be the reason for the seasonal change of both compounds. To simulate such a sedimentation event, intact sediment cores were covered with suspensions of Porphyridium aerugenium or Synechococcus sp. The addition of the phytoplankton material resulted in a drastic increase of acetate concentrations with a maximum at 2 cm depth, similar to in situ acetate concentrations measured in summer. The same applies for CH₄ for which increased concentrations were observed down to 6 cm depth. H₂ concentrations, on the other hand, showed no distinct increase. Treatment of intact sediment cores with ¹⁴C-labeled Synechococcus cells resulted in the formation of ¹⁴C-acetate, ¹⁴CH₄ and ¹⁴CO₂. Maximum concentrations of ¹⁴CH₄ were found at 4 cm depth, i.e. just above the depth to which ¹⁴C-acetate penetrated. The results show that phytoplankton blooms may cause a seasonal variation of acetate and CH₄ in profundal sediments of deep lakes despite the constant low temperature. They also indicate that acetate is the dominant substrate for methanogenic bacteria in the profundal sediments of Lake Constance.     

Differential Effects of 4'-Chlorodiazepam on Expressed Human GABAA Receptors

Abstract: The interactions of the atypical benzodiazepine 4'-chlorodiazepam (Ro 5-4864) with functionally expressed human GABA_A receptor cDNAs were determined. Cotransfection of human α₂, β₁, and γ₂ subunits was capable of reconstituting a 4'-chlorodiazepam recognition site as revealed by a dose-dependent potentiation of t -[³⁵S]butylbicyclophosphorothionate ([³⁵S]TBPS) binding to the GABA-activated chloride channel. This site is found on GABA_A receptor complexes containing sites for GABA agonist-like benzodiazepines and neuroactive steroids. The importance of the α subunit was further demonstrated as substitution of either α₁ or α₃ for the α₂ subunit did not reconstitute a 4'-chlorodiazepam recognition site that was capable of modulating [³⁵S]TBPS binding under the same experimental conditions. The 4'-chlorodiazepam modulatory site was shown to be distinct from the benzodiazepine site, but the phenylquinolines PK 8165 and PK 9084 produced effects similar to 4'-chlorodiazepam, consistent with the previous analysis of the 4'-chlorodiazepam site in brain homogenates. Further analysis of the subunit requirements revealed that coexpression of α₂ and β₁ alone reconstituted a 4'-chlorodiazepam recognition site. It is interesting, however, that the 4'-chlorodiazepam site was found to inhibit [³⁵S]TBPS binding to the GABA-activated chloride channel. Thus, the 4'-chlorodiazepam site may be reconstituted with only the α and β polypeptides.     

Chernobyl-derived radiocaesium in fish as dependent on water quality and lake morphometry

¹³⁷Cs concentrations in perch Perca fluviatilis , pike Esox lucius and roach Rutilus rutilus obtained from lakes of different size and water quality in an area which received about 10–67 kBq m⁻²¹³⁷Cs, were compared with environmental data. Radiocaesium concentrations were highest in pike, and were about two to three times higher in the pike and perch than in the roach. The largest perch had about four times more ¹³⁷Cs than the smallest ones, but the activities in the pike and roach were independent of fish size. All of the water quality parameters examined correlated with ¹³⁷Cs concentrations in the fish, but the concentrations of ¹³⁷Cs in the bottom sediment did not. ^l37Cs fallout and water retention time in lakes showed a positive correlation with fish ¹³⁷Cs, while the size of the catchment area, phosphorus content of the water, water colour, pH and electrical conductivity showed a negative one. Thus there was more ¹³⁷Cs in the fish from the oligotrophic lakes than from the eutrophic ones, from the less humic lakes than from the more humic ones, from the more acid than from the less acid ones and from the lakes containing less potassium than from those containing more potassium. The depth, area and volume of the lake did not affect radiocaesium in the fish. Regression equations are presented for predicting ¹³⁷Cs in fish on the basis of environmental data. Fallout, total phosphorus and pH together had the highest influence in multiple regression models but were less significant in the roach than in the perch and pike.     

L-GLUTAMIC ACID DECARBOXYLASE IN NON-NEURAL TISSUES OF THE MOUSE

Abstract— Low levels of γ-aminobutyric acid (GABA) and of glutamic acid decarboxylase (GAD) activity have been detected in mouse kidney, liver, spleen and pancreas. Quantitation of both ¹⁴CO₂ and [¹⁴C]GABA produced in radiometric assays from [U-¹⁴CJglutamic acid has shown that measurement of ¹⁴CO₂ evolution alone is not, in all cases, a valid estimate of true GAD activity. As evidenced by increased ^,14CO₂ production upon addition of NAD and CoA to assay mixtures, radiometric assay of GAD activity in crude homogenates may yield ¹⁴CO₂ via the coupled reactions of glutamic acid dehydrogenase and a-ketoglutarate dehydrogenase. The addition of 1 mM aminooxyacetic acid (AOAA) to assays of kidney homogenates inhibited [^,14C]GABA production 92 per cent while ¹⁴CO₂ production was inhibited only 53 per cent. No evidence was found to confirm the reported existence of a second form of the enzyme, GAD II. previously described by Haber el al. (H aber B., K uriyama K. & R oberts E. (1970) Biochem. Pharmac. 19, 1119-1136). Based on sensitivity-to AOAA and chloride inhibition, the GAD activity in mouse kidney is. apparently, indistinguishable from that of neural origin.     

Stimulation of shoot elongation in Salix pentandra by gibberellin A9; activity appears to be dependent upon hydroxylation to GAl via GA20

Cessation of shoot elongation in seedlings of Salix pentandra L. is induced by short photoperiod. Gibbereliin A₉ (GA₉) applied either to the apical bud or injected into a mature leaf, induced shoot elongation under a short photoperiod of 12 h, and GA₉ could completely substitute for a transfer to a long photoperiod. When [³H]GA₉ or [²H₂]GA₉ was injected into a leaf, no [³H]GA₉ was detected in the elongating apex and only traces of [³H]GA₉ were found in the shoot above the treated leaf. By the use of gas chromatography-mass spectrometry (GC-MS), [²H₂]GA₂₀ was identified as the main metabolite of [²H₂]GA₉ in both the shoot and the treated leaf. In addition, [²H₂]GA₁ and [²H₂]GA₂₉ were also identified as metabolites of [²H₂]GA₉. These results are consistent with the hypothesis that exogenous GA, promotes shoot elongation in Salix through its metabolism to GA₂₀ and GA,.     

Gas exchange measurements required to predict the newly fixed carbon pool size in a source leaf of corn (Zea mays L.)

Abstract. Steady-state photosynthesis (P_n), post-illumination CO₂ release rates (R), sucrose-phosphate synthase (SPS) activities, and levels of starch, sucrose and hexoses were measured in the source leaf of corn ( Zea mays L.) during a 16-h photoperiod at 800 μmol m ² s ¹. P_n and SPS activity remained constant. Carbohydrate pools increased at a linear rate, except the first and last hour of the photoperiod. Both the CO₂ evolution rate at the moment of light removal (R_max) and SPS activity decreased by one half after the onset of darkness (0 60 min). Sucrose diminished during this period by 40%, whereas the starch remained constant. Thereafter, starch mobilization began, followed by a gradual decline in leaf respiration. The average dark export rate was calculated to be 60% less than that during the day. Maintenance respiration (R_m) of an attached leaf after 48 h darkness was determined. Plants were illuminated for different intervals (e.g. 5, 10 or 20 min), each followed by dark periods sufficient for respiration to decline to R_m. The ratio of C assimilated in light to that released in dark was 6:1. After the 48-h dark period, the minimal period of illumination (T_min) required to restore P_n and R_max to the original level was determined. A mathematical analysis of the kinetics involved in the recovery of P_n and R_max provided an estimate of turnover time (0.22h) and pool size 9.15 mmol m ²) for the newly fixed carbon.     

Dopamine Agonist-Mediated Inhibition of Acetylcholine Release in Rat Striatum Is Modified by Thyroid Hormone Status

Abstract: K⁺-evoked acetyl[³H]choline ([³H]ACh) release was inhibited in a concentration-dependent manner by apomorphine and the D₂ agonist quinpirole in striatal slices prepared from euthyroid and hypothyroid rats. However, there was a significant increase in the maximum inhibition observed with both agonists in the hypothyroid compared with the euthyroid group, which paralleled the increased D₂ agonist sensitivity reported for stereotyped behavior. The D₂ antagonist raclopride decreased, and the D, antagonist SCH 23390 increased, the inhibition of [³H]ACh release by apomorphine, confirming an inhibitory role for D₂ receptors and an opposing role for D₁ receptors. Because there is no difference in D₁ or D₂ receptor concentration between the euthyroid and hypothyroid groups, it is suggested that thyroid hormone modulation of D₂ receptor sensitivity affects a receptor-mediated event. Following intrastriatal injection of pertussis toxin (PTX), apomorphine no longer inhibited [³H]ACh release. In fact, increased [³H]- ACh release was observed, an effect reduced by SCH 23390, providing evidence that D₁ receptors enhance [³H]- ACh release, and confirming that a PTX-sensitive G protein mediates the D₂ response. As it has been reported that thyroid hormones modulate G protein expression, this mechanism may underlie their effect on dopamine agonist- mediated inhibition of ACh.     

Distinct Loci Mediate the Direct and Indirect Actions of the Anesthetic Etomidate at GABAA Receptors

Abstract: Most general anesthetics produce two distinct actions at GABA_A receptors. Thus, these drugs augment GABA-gated chloride currents (referred to as an indirect action) and, at higher concentrations, elicit chloride currents in the absence of GABA (referred to as a direct action). Because a β subunit appears to be required for the direct action of intravenous anesthetics in recombinant GABA_A receptors, site-directed mutagenesis of the β3 subunit was performed to identify amino acid residues that are critical for this action. In HEK293 cells expressing a prototypical GABA_A receptor composed of α1β3γ2 subunits, mutation of amino acid 290 from Asn to Ser dramatically reduced both etomidate-induced chloride currents and its ability to stimulate [³H]flunitrazepam binding. By contrast, the ability of etomidate to augment GABA-gated chloride currents and GABA-enhanced [³H]flunitrazepam binding was retained. The demonstration that the direct, but not the indirect, actions of etomidate are dependent on β3(Asn²⁹⁰) indicates that the dual actions of this intravenous anesthetic at GABA_A receptors are mediated via distinct loci.     

Chitinolytic bacteria and chitin mineralization in the marine waters and sediments along the Antartic Peninsula

Abstract Chitinolytic bacteria were enumerated and isolated from marine waters and sediments along the highly productive Antarctic Peninsula. Chitinolytic bacteria were found in low concentrations (approximately 1 cell per ml) in the water column and at much higher levels in marine surface sediments (10⁴–10⁵ per g). The predominant chitinolytic bacteria isolated from the water column were identified as psychrophilic Vibrio spp. Rates of chitin mineralization were measured by collection of ¹⁴CO₂ respired from ¹⁴C-labeled chitin synthesized from chitosan and [1-¹⁴C]acetic anhydride. Chitin mineralization rates were extremely low in the marine waters analyzed (0.00085–0.0019% of the added label respired in 48 h) and appreciably higher in the marine sediments (0.0039–0.01% per 48 h), suggesting that the sediments are much more important in chitin degradation. Such low mineralization rates suggest that chitin may be accumulating in Antarctic marine sediments, though animals may also play an important role in chitin degradation.     

STUDIES ON LIVER REGENERATION: I. 131IODODEOXYURIDINE AS A PRECURSOR of DNA IN NORMAL and REGENERATING RAT LIVER

Abstract. ¹³¹Iododeoxyuridine (¹³¹IDU) was injected into normal and partially hepatectomized rats, and the specificity of incorporation of this thymidine analogue into liver DNA was determined 2, 24 and 48 hr following intramuscular injection. At 2 and 24 hr after ¹³¹DU injection, a major proportion of radioactivity in the liver was in the acid-soluble fraction, whereas 48 hr after injection the label in the acid-soluble fraction had decreased considerably. In liver obtained 2 hr after injection of ¹³¹IDU, only 1.8–16.6% of the total radioactivity were in DNA. If, however, the tissue was subjected to formalin fixation, the acid-soluble label was extracted selectively, and of the remaining radioactivity 64–88% was in DNA. Therefore, the radioactivity that is not extracted by formalin may be used as a measure of DNA synthesis at the time of injection of ¹³¹IDU, thus obviating time-consuming biochemical fractionation procedures.