Growth dynamics of white myotomal muscle fibres in the bluntnose minnow, Pimephales notatus Rafinesque, and comparison with rainbow trout, Salmo gairdneri Richardson

The dynamics of increase of the predominant white muscle fibres of the myotomal bulk in bluntnose minnow, Pimephales notatus , ranging from 2.0 to 9.1 cm f.l. have been analysed by examination of modal progression of fibre diameter frequency classes in fish fed to satiation and growing at different rates at 15, 25 or 30°C. Recruitment of new fibres appeared to contribute little to increase in muscle bulk above 4 cm f.l. , and nothing beyond 6 cm. The dominant means of increase was increase in fibre diameter. The limiting fibre diameter seemed to be 120 μ. These dynamics, which result in an approximately 1: 1 ratio between mean fibre diameter and f.l. , are in contrast to those of the myotomal white muscle of rainbow trout, Salmo gairdneri , in which, regardless of differences in somatic growth produced by temperature, ration size or growth hormone administration, mean fibre diameter does not exceed that in bluntnose minnow until trout exceed 30 cm f.l. In trout there is, moreover, input of new fibres up to approximately 50 cm f.l. , when subsequent growth, as in the minnow, is by means of fibre diameter increase. The bluntnose minnow is a small, slow growing species; the rainbow trout is a large, fast growing species. The discussion links these facts with the observed differences in fibre growth dynamics in relation to a hypothesis of interspecific differences in fish growth capability.     

The relationship between mosaic muscle fibres and size in rainbow trout (Salmo gairdneri)

The dynamics of increase of the mosaic muscle in hatchery reared rainbow trout (2·3 to 61·3 cm fork length) are investigated. In trout <5 cm, all fibres are <40 μm in diameter; from 5 to 20 cm the diameter mode remains in the 0–39·9 μm class and there is some extension in range of diameter, thereby suggesting that mosaic muscle increase up to 20 cm is mainly by recruitment of new (small) fibres. When trout exceed 20 cm, mode of fibre diameter shifts to the 40–79·9 um class and fibres of larger diameter (> 100 μrn) appear but the subsequent overall fibre diameter frequency distribution changes little until 50 cm. Increase in muscle during the phase 20–50 cm seems partly attributable to increase in fibre diameter, but remains largely the result of recruitment of small fibres, although the mechanism of the latter process appears less clear than in fish of <20 cm. The recruitment of new fibres stops at 55 cm and further increase in mosaic muscle evidently depends upon the ability of existing fibres to increase in diameter. Possible means by which increasing muscle fibre diameter may limit the ultimate size which trout may achieve are discussed. A phenomenon of apparent reduction in fibre diameter in winter among fish 20–39·9 cm long is noted and its significance is considered.     

Postlarval muscle growth in fish: a DNA flow cytometric and morphometric analysis

The mechanism of postlarval fish myotomal growth was investigated in trout (Salmo gairdneri) by means of morphometric and cytofluorometric analysis. The mechanism by which new fibres are added during postlarval growth (hyperplasia) is not fully understood. In histological cross sections these new fibres have a small diameter which give the muscle a "mosaic" appearance. One hypothesis suggested that they could be derived from the proliferative activity of satellite cells. DNA cytofluorometric analysis of nuclei suspensions obtained from trout white myotomal muscle during different developmental stages (eleutherembyronic; alevin; yearling and adult) showed a consistently low S-cytometric phase during all stage in which myofibres of small diameters were present. The percentage of such small fibres, determined by morphometric analysis, suggested that satellite cells are the proliferative population. In fact, their percentages, as determined by morphometric analysis in histological section, bear a linear relationship with the S-cytometric phase percent nuclei (R = 0.927). Only in adults (67 cm in size) there was a significant decrease in the S-cytometric phase. At this stage, in histological sections, the myotomal muscle no longer had a "mosaic" appearance because of the disappearance of the small fibres. It may, therefore, be supposed that in the cm 67 adult specimens, the proliferative population is entering the G0 phase. It is known, in fact, that muscle growth proceeds only by fibre hypertrophy in trout longer than 70 cm in length (Stickland, 1983).     

Muscle fibre growth dynamics in diploid and triploid rainbow trout

The effect of triploidy on muscle fibre growth was determined by comparing hyperplasia and hypertrophy of white muscle fibres in all-female, diploid and triploid rainbow trout Oncorhynchus mykiss (100–400 mm total length). Conventional morphometry and protein and DNA concentrations were used to assess muscle fibre hyperplasia and hypertrophy in white muscle samples derived from an anterio-dorsal location. Muscle fibre distributions were significantly different between triploids and diploids in trout <300 mm. The proportion of fibres <20 μm was higher in diploids than in triploids and the proportion of fibres in the 20–40 μm category was higher in triploids than in diploids. This indicates that the hyperplastic fibres of triploids are larger than those of diploids. Larger hyperplastic fibres in triploids are probably due to the combined effect of increased nuclear size in triploids and the relatively high nucleus: cell ratio observed in small muscle fibres. These larger fibres may be less favourable to cellular metabolic exchange because of their smaller surface area to volume ratios, and perhaps account for reduced viability and growth observed in triploids during early life stages. On the other hand, the lack of difference in the distribution of fibres <20 μm between diploids and triploids at larger body size ranges (301–400 mm) imply that triploid trout may have higher rates of new fibre recruitment and growth capacity at these sizes. There was no difference between diploid and triploid trout in the mean size of muscle fibres; however, the number of fibres per unit area was reduced by 10% in triploids. No differences were observed in protein or DNA concentrations in muscle tissues between the two genetic groups. Since triploid nuclei have 1·5 times more DNA than diploid nuclei, this deviation from the expected muscle DNA concentration (1·3–1·4 times more DNA in triploids when the 10% reduction in fibre density is considered) suggests that the number of nuclei per muscle fibre is reduced. In both diploids and triploids, mean fibre size increased with body length while fibre density decreased. Similarly, protein concentration in the muscle tissue increased and DNA concentration declined with increasing body length. Protein/DNA ratio was strongly and positively correlated with fibre size. These results demonstrate that changes in DNA and protein concentrations can be used to assess hyperplasia and hypertrophy in muscle tissues. However, the morphometric procedure provides better insight into muscle fibre growth as it enables the direct visualization and analysis of muscle fibre distribution patterns.     

Influence of growth rate on white muscle dynamics in rainbow trout and brook trout

In trout, fast growth stimulated white muscle fibre hypertrophy ( P 0·001) and hyperplasia ( P <0·01) in outer fibres but not in deep fibres. Glycogen was most prevalent in outer fibres ( P <0·01) and in brook trout ( P <0·01) that on average had three to four times larger fibres than rainbow trout.     

Relative growth of tissues at different somatic growth rates in rainbow trout Salmo gairdneri Richardson

Growth of tissues (heart, spleen, liver, gonad, gut, skin, visceral fat and carcase) in immature rainbow trout relative to growth of whole fish, was examined by means of allometric analysis (y=ax^b; Huxley, 1932) of wet and dry weights. Relative growth rates of tissues are compared in an initial sample of fingerlings (< 11·29 g), and in fish (> 11·29 g) growing rapidly (at 12°C ad libitum rations) and slowly (at 12°C, 4–5% rations, and 7°C ad libitum rations). In the fingerlings of the initial sample, the major tissues by weight (liver, gut, skin, visceral fat) increased relatively more rapidly than whole body weight (i.e. with positive allometry; b> 1·00), and carcase increased more slowly than body weight (b< 1·00). Above fingerling size, the reverse holds: tissues other than carcase (with exception of visceral fat) increased less rapidly than body weight (b < 1·00), and carcase more rapidly (b > 1·00). These principles hold for post-fingerling, immature rainbow trout, regardless of growth rate differences of whole fish, and even in fish that also received bGH (bovine growth hormone). The characteristic b (slope) values in the allometry equations for the wet and dry weights of the various tissues tend to remain constant, despite whole fish growth rate differences and the effects of this constancy are to maintain approximate constancy of body proportional (%) wet and dry weights. It is emphasized however that the effect of b values only slightly > or < 1·00 can cause important changes in the proportional (%) weights of major tissues as fish continue to increase in size. In low (4–5%) rations fish (without bGH) the slope (b) value for gut is less than for fish on ad libitum rations with the result that gut proportional weight is significantly less among these fish. In fish on ad libitum rations at 7°C, the slope (b) value for skin is greater than for all other (12°C) groups; skin proportional weight for the 7°C group is therefore, significantly greater. The overall impression of relative growth in immature rainbow trout is of remarkable conservativeness of body proportional weights regardless of overall somatic growth rate.     

Growth dynamics of white and red muscle fibres in fast- and slow-growing strains of rainbow trout

Compared with fish of a slow-growing strain, fast-growing rainbow trout exhibited significantly smaller white fibre diameters, throughout development from hatching to 24 cm body length, although possessing similar total number of fibres. In contrast, in red muscle, no differences were observed in fibre diameter between the two strains, but the fast growing fish showed a significantly higher number of red fibres. The differences in growth rate between the two strains were related to the mean white fibre diameter and were found to be matched by proportional adjustments in recruitment of new fibres to the growing muscle. Thus, the largest and fastestgrowing strain showed evidence of sustained higher recruitment of muscle fibres that endowed this strain with the potential to maintain rapid somatic growth for longer and accomplish greater muscle growth.     

Sustained swimming speeds and myotomal muscle function in the trout, Salmo gairdneri

Rainbow trout were trained for 3–4 weeks in a flume at swimming speeds of 1, 2 and 3 l s⁻¹. For each experiment growth rates were estimated and by measuring the hypertrophy of red and mosaic skeletal muscle fibres their function was described at particular swimming speeds and compared with earlier experiments on coalfish using the same technique.
Maximum growth, compared with controls in still water, occurred at swimming speeds of 1 l s⁻¹. At this speed the trout mosaic muscle fibres hypertrophied by 40% but the red muscle fibres showed only a 25% hypertrophy. It is suggested that natural swimming speeds are close to 1Ls^−l and the trout mosaic fibres are better adapted for use at this speed in comparison with coalfish white muscle fibres.     

Recruitment and maximal diameter of axial muscle fibres in teleosts and their relationship to somatic growth and ultimate size

Growth of white axial muscle fibres of ten species of freshwater teleosts from five families (Cyprinidae, Centrarchidae, Percidae, Salmonidae, Esocidae) possessing widely different growth rates and ultimate sizes have been studied. The dynamics of muscle increase (i.e. increase in fibre numbers and/or diameter) appears to determine the ability for rapid somatic growth and large ultimate size in teleosts. Thus, the largest and fastest growing species (smallmouth bass, lake whitefish, rainbow trout, muskellunge) show evidence of sustained recruitment of muscle fibres to a large size, in contrast to the smaller and slower growing species (bluntnose minnow, longnose dace). Pumpkinseed, bluegill and yellow perch are all intermediate in fibre growth dynamics, growth and ultimate size between the smaller and larger species. Moreover, the ability of teleosts to grow rapidly and attain a large ultimate size is dependent on the body length at which recruitment of new muscle fibres into the growing axial muscle ceases. The regression equation, y =– 0.29 + 2.26 ( x ), showing the relationship (r = 0.95) between ultimate body length, x , and fork length at cessation of recruitment, y , indicated that for these teleosts, recruitment tends to cease when the fork length reaches about 44% of the ultimate body length. Possible mechanisms to account for this relationship are proposed, and the role of the ultimate fibre diameter in posing limits to the ultimate size of the species is discussed.     

Growth dynamics of caudal and pectoral fin muscle fibres in a carangid, Caranx malabaricus (Cuv. & Val.), and their possible relation with somatic growth

The growth dynamics of red, pink and white fibres of the caudal and pectoral fin muscles are described in Carans malabaricus (Cuv. & Val.) in relation to their somatic growth. In all three fibre types growth occurred by an increase in fibre number and diameter in small size classes of fish and by an increase in diameter only in larger fish. The growth dynamics of the three fibre types were similar to those of the myotomal muscle fibres and paralleled the somatic growth pattern of this fish.     

Skeletal muscle afferent regulation of bioassayable growth hormone in the rat pituitary

There are forms of growth hormone (GH) in theplasma and pituitary of the rat and in the plasma of humans that areundetected by presently available immunoassays (iGH) but can bemeasured by bioassay (bGH). Although the regulation of iGHrelease is well documented, the mechanism(s) of bGH release is unclear.On the basis of changes in bGH and iGH secretion in rats that had been exposed to microgravity conditions, we hypothesized that neural afferents play a role in regulating the release of these hormones. Toexamine whether bGH secretion can be modulated by afferent input fromskeletal muscle, the proximal or distal ends of severed hindlimb fastmuscle nerves were stimulated (~2 times threshold) in anesthetizedrats. Plasma bGH increased ~250%, and pituitary bGH decreased~60% after proximal nerve trunk stimulation. The bGH response wasindependent of muscle mass or whether the muscles were flexors orextensors. Distal nerve stimulation had little or no effect on plasmaor pituitary bGH. Plasma iGH concentrations were unchanged afterproximal nerve stimulation. Although there may be multiple regulatorymechanisms of bGH, the present results demonstrate that the activationof low-threshold afferents from fast skeletal muscles can play aregulatory role in the release of bGH, but not iGH, from the pituitaryin anesthetized rats.

     

Functional expression of bovine growth hormone gene in Pleurotus eryngii

The expression of the bovine growth hormone (bGH) gene was examined in Pleurotus eryngii, which belongs to the family of oyster mushrooms. The region encoding mature bGH, which has a variety of regulatory effects on growth and metabolic processes, was amplified using designed primers containing initiation and termination codons and then subcloned into pPEV binary expression vector. The recombinant vector (pPEVbGH) was introduced in P. eryngii via Agrobacterium tumefaciens-mediated transformation. Recombinant bGH was expressed in P. eryngii harboring pPEVbGH vector under control of the cauliflower mosaic virus (CaMV) 35S promoter up to a level of approximately 26% of total cell proteins after 6 days of cultivation, after which the recombinant protein was analyzed by SDS-PAGE and Western blotting. Interestingly, the growth rate of P. eryngii mycelia harboring pPEVbGH vector was approximately three times faster than that of control P. eryngii, suggesting that bGH affected the growth of P. eryngii. Biological activities were examined in Sprague-Dawley rats, which were administered regular feed mixed with mycelial extracts containing bGH (0.1 or 0.2 μg of bGH per g of animal feed). Mycelial extracts containing bGH significantly affected growth rates and lipid profiles; total cholesterol, triglyceride, HDL, and LDL levels were improved in rats fed mycelial extracts compared with those administered regular feed containing nontransgenic P. eryngii. This result indicates that P. eryngii harboring pPEVbGH vector could produce biologically active bGH. Further, levels of all growth-related factors increased, resulting in faster growth rates in bGH-treated groups. Accordingly, these data suggest that P. eryngii can be applied to the production of industrially useful proteins using a plant expression vector as an efficient mushroom host system.     

Growth and multiplication of white skeletal muscle fibres in carp larvae in relation to somatic growth rate

A morphometric analysis of white axial muscle of common carp Cyprinus carpio was undertaken in order to quantify increase in fibre size, fibre nuclei and fibre number in relation to somatic growth rate during early life. In fast-growing carp larvae fed zooplankton, length and height of fibres from the central part of dorsolateral muscle increased at the same rate (0.75) relative to the total length of the larvae during the first 2 weeks of feeding. During this period, the number of nuclei per fibre increased threefold while the number of nuclei per unit fibre surface remained constant. In fast-growing larvae fed a formulated diet, the total cross-sectional area of one epaxial quadrant of white muscle and the total area of white fibres increased at almost the same rate (3.15; 3.23) relative to larval total length during the first 28 days of exogenous feeding. The total number of white fibres increased faster (2.07) relative to the total length of larvae than the mean area of white fibres (1.16). Hyperplasia accounted for 64% of muscle growth in these larvae. The proportion of fibres with a width < 10 μm decreased from 72% at first feeding to 14% 28 days later, while the proportion of fibres with a width >20 μm which was 0% at first feeding increased up to 34% in the same time. The recruitment of new white fibres seemed to be almost the same in the whole muscle quadrant at first feeding and 18 or 28 days later but, 8 days after first feeding, a transient significant recruitment of new fibres was shown at the apex of the myotome. Comparisons between fast- and slow-growing groups of larvae showed that for a given larval total length: (1) the mean width of central white fibres was higher and the proportion of central fibres with a width <10 μm was lower in slow-growing larvae than in fast-growing ones; (2) the total number of white fibres was lower for a higher total cross-sectional area of white muscle in slow-growing larvae than in fast-growing ones. These results suggest that, in Cyprinus carpio larvae, slow-growing conditions are related to a decreased contribution of hyperplasia to muscle growth.     

Regional variations in fibre growth dynamics of myotomal and caudal fin muscles in relation to body size of a freshwater teleost, Barbus sarana (Cuv. & Val.)

The growth of red fibres in anterior and middle myotomal regions of B. sarana was mainly by hyperplasia in smaller size classes. In higher size classes, growth by hyperplasia was greater in posterior myotomal region compared to the other two myotomal regions. The growth of pink fibres in anterior myotomal regions was mainly by hypertrophy. The middle and posterior myotomal regions showed fibre growth by hyperplasia. The growth dynamics of white fibres revealed more or less similar pattern in all three myotomal regions against the somatic development. White fibres grew by hyperplasia up to 8 cm F.L. size classes and thereafter by hypertrophy. However, in > 12 cm F.L. size classes, the mean diameter of white fibres did not increase significantly. Similar pattern of growth was found in the white fibres of caudal fin muscle. It is interesting to note that the hyperplasia was mostly completed in the white fibres of the smallest fish studies, whereas, it continued to quite larger fish size in red and pink fibres. Thus, hyperplasia and hypertrophy may be responsible for growth in all fibre types in all myotomal regions in relation to somatic development in this small and medium growing species.     

Metabolic effects of developmental, tissue-, and cell-specific expression of a chimeric phosphoenolpyruvate carboxykinase (GTP)/bovine growth hormone gene in transgenic mice.

Transgenic mice were used to investigate sequences within the promoter of the gene for the cytosolic form of phosphoenolpyruvate carboxykinase (GTP) from the rat (EC 4.1.1.32) (PEPCK) which are involved in tissue-specific and developmental regulation of gene expression. Segments of the PEPCK promoter between -2000 and -109 were linked to the structural gene for bovine growth hormone (bGH) and introduced into the germ line of mice by microinjection. Bovine growth hormone mRNA was found in tissues that express the endogenous PEPCK gene, mainly in the liver but to a lesser extent in the kidney, adipose tissue, small intestine, and mammary gland. In the liver the chimeric PEPCK/bGH(460) gene was expressed in periportal cells, which is consistent with the zonation of endogenous PEPCK. The PEPCK/bGH gene was not transcribed in the livers of fetal mice until immediately before birth; at birth the concentration of bGH mRNA increased 200-fold. Our results indicate that the region of the PEPCK promoter from -460 to +73 base pairs contains regulatory sequences required for tissue-specific and developmental regulation of PEPCK gene expression. Mice transgenic for PEPCK/bGH(460) were not hyperglycemic or hyperinsulinemic in response to elevated bGH, as were transgenic mice with the MT/bGH gene. The number of insulin receptors in skeletal muscle was no different in mice transgenic for MT/bGH when compared with mice transgenic for PEPCK/bGH(460) and control animals. However, mRNA abundance for the insulin-sensitive glucose transporter in skeletal muscle was decreased in mice transgenic for the MT/bGH gene. The differences in glucose homeostasis noted with the two types of transgenic mice may be the result of the relative site of expression, the different developmental pattern, or hormonal regulation of expression of the bGH gene.     

Changes in muscle structure associated with somatic growth in Idiosepius pygmaeus, a small tropical cephalopod

Mantle muscle tissue of Idiosepius pygmaeus was examined to describe changes in structure and organization associated with growth. Growth in I. pygmaeus|DD was a function of both an increase in muscle fibre number and fibre size within muscle blocks. Continuous fibre production over the observed life span of I. pygmaeus was indicated by the presence of very small muscle fibres (< 1.0 μm in diameter) in substantial proportions in all sizes of individuals. Muscle blocks became larger as animals increased in size, although new muscle blocks were generated in all sizes of individuals. Mantle muscle fibres had a maximum size of 11 μm. Therefore, for an individual to continue increasing muscle block sizes, new fibres must be produced. This is further evidence of continuous fibre production throughout the size range of I. pygmaeus examined. The relative rates of muscle fibre generation and fibre growth depended on the size of the animal and position along the mantle (anterior, mid or posterior mantle). The predominance of small fibres and blocks at the anterior end of the mantle suggested that this was the primary growth region. Mitochondriapoor and mitochondria-rich muscle fibres from small individuals had much larger mitochondrial cores than muscle fibres from larger animals. Changes in the muscle structure are discussed with respect to the metabolic and energetic requirements of I. pygmaeus , and how these may change with growth.     

Tissue deposition rates in relation to muscle fibre and fat cell characteristics in lean female pigs (Sus scrofa) following treatment with porcine growth hormone (pGH)

Two experiments were conducted to examine the effect of porcine growth hormone (pGH) on performance, carcase composition, muscle and fat deposition rates, muscle fibre characteristics, and fat cell volume in pigs. In the first experiment, ten pairs of littermates were treated with vehicle (saline buffer) or 80 μg pGH per kg live weight per day for 42 days starting at 50 kg live weight. In the second experiment, twelve pairs of littermates were untreated or treated with 3 mg pGH per day from approximately 56 kg live weight to slaughter at approximately 103 kg live weight. All pigs were fed ad libitum. In Experiments 1 and 2, respectively, feed intake decreased (10 and 11%) and the feed/gain ratio improved (8 and 13%), while daily gain was unaltered. There was an increase in deposition rates of muscle (11 and 22%), skin (27 and 23%), and bone (15% in both), and a decrease in deposition rates of intermuscular (48 and 24%) and subcutaneous (82 and 50%) fat. This resulted in a change in carcase composition towards more muscle (5 and 9%), bone (6 and 4%), and skin (18 and 12%), and less intermuscular (30 and 16%) and subcutaneous fat (51 and 32%). The increased muscle mass was due to enhanced hypertrophy of all muscle fibre types, while pGH did not affect the frequency of the different muscle fibre types. The reduction in subcutaneous fat was reflected in a similar reduction in fat cell volume. In contrast to the majority of pigs used in pGH experiments, the genotype used in the present experiments did not respond with respect to daily gain following pGH treatment. Furthermore, the increase in muscle deposition was rather low compared to results reported in pigs of other genotypes. These data together with published data on the cross-sectional area of muscle fibres indicate that genotypes with relatively large muscle fibres are less responsive to pGH treatment than genotypes with relatively small muscle fibres.     

Quantitative determination of growth hormone by immunoblotting

Blotting techniques have been extensively used, not only analytically for protein identification, but also preparatively to isolate and purify specific proteins from a large variety of cellular extracts and biological fluids. The process involves the separation of proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transfer of proteins to nitrocellulose membranes, and immunostaining to identify proteins which often are at very low concentrations. Because of the quantitative interactions of the proteins with specific antibodies, we have coupled the immunoblot procedure with photographic and densitometric methods for the quantitative determination of bovine growth hormone (bGH). In this way, the method is suitable for bGH detection and quantitation for a small number of samples by use of a single Western blot analysis. The sensitivity of this method permits determinations of bGH to 0.5 ng. The method uses a comparative procedure in which purified bovine growth hormone is used as a standard.     

Delayed appearance of liver growth hormone binding sites and of growth hormone-induced somatomedin production during rat development

The present study was designed to determine whether the apparent paradox of high circulating growth hormone levels in the fetus and the minimal effect of this hormone on growth might reflect a diminished responsiveness of fetal target organs to GH. Specific uptake by rat liver of [125I] bGH was very low in fetuses as compared to suckling and adult rats. Also, liver uptake of the iodinated hormone decreased proportionally with the simultaneous injection of increasing amounts of growth hormone, but was not modified by the simultaneous injection of unlabelled chemically-related hormones. Since the water content is significantly greater in fetal than adult tissues, results were expressed by liver dry weight and again, [125I] bGH liver uptake continued to increase with age. After bovine growth hormone administration to adult rats, plasma somatomedin C concentrations increased significantly, while they had no effect in fetuses. These results suggest that reduced liver somatogenic binding sites in the fetus prevents growth hormone from inducing growth-promoting effects during intrauterine life.     

RNA-DNA ratio as an index to growth in salmonid fishes in the laboratory and in streams contaminated by carbaryl

Free nucleotides, RNA, and DNA were extracted from fish white muscle and analysed by a two-wavelength ultraviolet (UV) absorbance technique to determine their relation to growth rates in brook trout, Salvelinus fontinalis , and Atlantic salmon, Salmo salar . In the laboratory, fish given ample feed had higher growth rates than fish on restricted rations, and these growth rates were significantly correlated with RNA-DNA ratios. Wild brook trout from natural streams had higher average RNA-DNA ratios than those in the laboratory. Exposure of brook trout in one of the streams to low levels of the insecticide carbaryl (Sevin) that drifted from the spraying of nearby forests did not reduce RNA-DNA ratios.