Thyroid hormones in growth and development of fish.

The thyroid hormones (THs), thyroxine (T(4)) and triiodothyronine (T(3)) are products of the thyroid gland in all vertebrates. Their role in early development and metamorphosis is well established in mammals and amphibians, respectively, and recently several studies in fish have highlighted the importance of THs during flatfish metamorphosis. THs are present in high quantities in fish eggs and are presumably of maternal origin. During embryogenesis the concentration of T(4) and T(3) in the eggs decrease until endogenous production starts. Thyroid hormone receptors (TR) have been isolated from several teleosts and in common with tetrapods two receptor isoforms have been identified, TR alpha and TR beta. Both the receptors are expressed in early embryos and larvae of the Japanese flounder (Paralichthys olivaceus), zebrafish (Danio rerio) and seabream (Sparus aurata) although a different temporal pattern is apparent. The role of THs and TRs in fish embryogenesis, larval development and during metamorphosis will be discussed.

Dopaminergic innervation of the rainbow trout pituitary and stimulatory effect of dopamine on growth hormone secretion in vitro.

To elucidate which factors regulate growth hormone (GH) secretion in rainbow trout, dopaminergic innervation of the rainbow trout pituitary along with the action of dopamine in vitro, were studied. Brains with attached pituitaries were double-labeled for putative dopaminergic neuronal fibers and somatotropes, using fluorescence immunohistochemistry. A direct dopaminergic innervation to the proximal pars distalis (PPD) with dopaminergic fibers terminating adjacent to somatotropes was demonstrated. Growth hormone secretion from whole pituitaries was measured in perifusate using a homologous GH-RIA. Dopamine (DA; 10(-7)-2x10(-6) g ml(-1)) increased basal GH secretion, with the GH secretion normalizing again after the DA exposure was halted. When pituitaries were pre-treated with somatostatin-14 (SRIF-14; 10(-12)-10(-9) g ml(-1)), before being exposed to different doses of DA, there was an inhibition of GH secretion which was not reversed after treatment of SRIF-14 was halted, unless DA was added. It is concluded that dopamine can function as a GH secretagogue in the rainbow trout pituitary gland.

The presence of high-affinity, low-capacity estradiol-17beta binding in rainbow trout scale indicates a possible endocrine route for the regulation of scale resorption.

High-affinity, low-capacity estradiol-17beta (E(2)) binding is present in rainbow trout scale. The K(d) and B(max) of the scale E(2) binding are similar to those of the liver E(2) receptor (K(d) is 1.6 +/- 0.1 and 1.4 +/- 0.1 nM, and B(max) is 9.1 +/- 1.2 and 23. 1 +/- 2.2 fmol x mg protein(-1), for scale and liver, respectively), but different from those of the high-affinity, low-capacity E(2) binding in plasma (K(d) is 4.0 +/- 0.4 nM and B(max) is 625.4 +/- 63. 1 fmol x mg protein(-1)). The E(2) binding in scale was displaced by testosterone, but not by diethylstilbestrol. Hence, the ligand binding specificity is different from that of the previously characterized liver E(2) receptor, where E(2) is displaced by diethylstilbestrol, but not by testosterone. The putative scale E(2) receptor thus appears to bind both E(2) and testosterone, and it is proposed that the increased scale resorption observed during sexual maturation in both sexes of several salmonid species may be mediated by this receptor. No high-affinity, low-capacity E(2) binding could be detected in rainbow trout gill or skin.

Growth hormone inhibits growth hormone secretion from the rainbow trout pituitary in vitro.

Growth hormone (GH) secretion in salmonids and other fish is under the control of a number of hypothalamic factors, but negative feed-back regulation by circulating hormones can also be of importance for the regulation of GH secretion. Mammalian studies show that GH has a negative feed-back effect on its own secretion. In order to elucidate if GH levels present a direct ultra-short negative feedback loop at the pituitary level GH secretion was studied in intact pituitaries from 50 g fish in an in vitro perifusion system. Following an initial equilibrium period pituitaries were exposed to five increasing concentrations (1-1,000 ng ml(-1)) of ovine GH (oGH) in 20-min steps, before being returned to a GH-free perifusion. Ovine GH caused a significant dose-dependent inhibition of GH secretion and it is concluded that GH can exert a direct negative feedback control on GH secretion at the pituitary level.

Photoperiod regulation of plasma growth hormone levels during induced smoltification of underyearling Atlantic salmon.

Earlier studies have established that increased daylength increases plasma growth hormone (GH) levels during spring smoltification of yearling Atlantic salmon. Recently, the Atlantic salmon aquaculture industry has started the production of underyearling ("summer") smolts. This involves fast juvenile growth on continuous light (24L), the transfer of juveniles over 8 cm in length to short day (12L) for 6 weeks in the summer, followed by transfer to 24L for another 6 weeks before transfer to seawater in late October. Three groups were studied in fresh water from July to the following May in order to elucidate the GH response to this photoperiod manipulation: one group was kept on 24L throughout (long-day group), while the other two groups were exposed to short day from July 15th. Of these, one was brought back onto long day on September 1st (winter group) while the other was kept on short day (short-day group). Plasma GH levels of the long-day group were around 1.6 ng/ml throughout the study. The short-day transfer suppressed GH levels to 0.7 ng/ml within 2 weeks (short-day and winter groups). The long-day transfer (winter group) increased GH levels to 11 ng/ml within 3 weeks, and this elevation of GH levels was sustained for about 3 months, before declining to pretreatment levels. The study demonstrates that underyearling Atlantic salmon react to increased daylength in a way similar to traditional yearling smolts. It also demonstrates for the first time that decreased daylength can suppress plasma GH levels in fish. It is concluded that winter photoperiod manipulation causes an out-of-season initiation and completion of the parr-smolt transformation of underyearling Atlantic salmon and that growth hormone plays a major role in this process.

Low temperature limits photoperiod control of smolting in atlantic salmon through endocrine mechanisms.

We have examined the interaction of photoperiod and temperature in regulating the parr-smolt transformation and its endocrine control. Atlantic salmon juveniles were reared at a constant temperature of 10 degrees C or ambient temperature (2 degrees C from January to April followed by seasonal increase) under simulated natural day length. At 10 degrees C, an increase in day length [16 h of light and 8 h of darkness (LD 16:8)] in February accelerated increases in gill Na(+)-K(+)-ATPase activity, whereas fish at ambient temperature did not respond to increased day length. Increases in gill Na(+)-K(+)-ATPase activity under both photoperiods occurred later at ambient temperature than at 10 degrees C. Plasma growth hormone (GH), insulin-like growth factor, and thyroxine increased within 7 days of increased day length at 10 degrees C and remained elevated for 5-9 wk; the same photoperiod treatment at 2 degrees C resulted in much smaller increases of shorter duration. Plasma cortisol increased transiently 3 and 5 wk after LD 16:8 at 10 degrees C and ambient temperature, respectively. Plasma thyroxine was consistently higher at ambient temperature than at 10 degrees C. Plasma triiodothyronine was initially higher at 10 degrees C than at ambient temperature, and there was no response to LD 16:8 under either temperature regimen. There was a strong correlation between gill Na(+)-K(+)-ATPase activity and plasma GH; correlations were weaker with other hormones. The results provide evidence that low temperature limits the physiological response to increased day length and that GH, insulin-like growth factor I, cortisol, and thyroid hormones mediate the environmental control of the parr-smolt transformation.

Growth hormone increases aggressive behavior in juvenile rainbow trout.

The aim of the present study was to clarify the role of growth hormone in social interactions in juvenile salmonids. Growth hormone increases the metabolic demands and feeding motivation in teleost fish. As a consequence, growth hormone may increase aggression levels and/or fighting ability. To test these hypotheses we observed agonistic behavior in pairs of juvenile rainbow trout (Oncorhynchus mykiss) consisting of two control fish (C/C pairs), two growth hormone-treated fish (GH/GH pairs), or one growth hormone-treated and one control fish (C/GH pairs). The initiator and the winner of each act of aggression were registered. Aggression was lowest in the C/C pairs, intermediate in the C/GH pairs, and highest in the GH/GH pairs, with the difference between the C/C pairs and the GH/GH pairs being significant. This supports the hypothesis that GH increases aggression levels. However, in the C/GH pairs, the number of conflicts won by GH-treated and control fish did not differ significantly. Thus, because social status was not increased, GH did not appear to affect fighting ability. We suggest that growth hormone affects aggression indirectly by increasing the swimming activity, and/or by inducing defense of a larger territory, thereby increasing the encounter rate between opponents. Since increased aggression can incur energetic and mortality costs, there may be selection against high GH levels in natural populations.

Cortisol inhibits glycosaminoglycan synthesis in cultured rainbow trout cartilage.

In vitro actions of corticosteroids (cortisol, 11-deoxycortisol, cortisone, and corticosterone) as well as interaction between cortisol and triiodothyronine (T3) or recombinant human insulin-like growth factor-I (rhIGF-I) on cartilage glycosaminoglycan (GAG) synthesis in rainbow trout (Oncorhynchus mykiss) were examined. Uptake of [35S]sulfate by isolated branchial cartilage was measured as a marker for GAG synthesis. In vitro exposure of cartilage to cortisol at concentrations of 10, 100, and 1000 nM for 6 days dose-dependently inhibited sulfate uptake, while exposure to 0.1 and 1 nM cortisol had no effect. Corticosterone and 11-deoxycortisol at concentrations of 10 and 100 nM inhibited sulfate uptake slightly but not dose-dependently. Cortisone (1, 10, and 100 nM) had no effect. When cortisol (1, 10, and 100 nM) and T3 (0.075 and 0.75 nM) were simultaneously added to the culture, the T3-induced sulfate uptake was dose-dependently reduced by the presence of 10 and 100 nM cortisol. When cortisol (1, 10, and 100 nM) and rhIGF-I (0.1 and 1 nM) were added together, the sulfate uptake induced by 0.1 nM rhIGF-I was only slightly inhibited by 100 nM cortisol, but 1 nM rhIGF-I completely masked the inhibitory effect of cortisol. These data suggest that GAG synthesis in the rainbow trout cartilage is controlled by multiple interactions among stimulative hormones, such as T3 and IGF-I, and inhibitory hormones, such as cortisol.

Growth hormone increases predation exposure of rainbow trout.

The energetic state of an animal strongly influences decisions that balances feeding against predation risk. Growth hormone increases the metabolic demands, which should elevate the feeding motivation of an animal. This, in turn, may increase the willingness to risk exposure to predators during feeding. To test this hypothesis, we studied the effect of growth hormone on the behavioural response of rainbow trout (Oncorhynchus mykiss) to simulated attacks from a model heron. After attacks, growth hormone treated trout foraged closer to the water surface, resumed feeding earlier, and ate more food than did control trout. Such behaviour should increase the susceptibility to aerial predation. Thus, predation may select against high endogenous growth hormone secretion in wild fish. Furthermore, genetic manipulations to increase growth hormone levels, intended to improve growth performance in aquaculture, may result in individuals with substantially altered behavioural patterns. In light of the increasing potential for interactions between farmed and wild fish, growth hormone transgenic fish may pose a threat to wild fish populations.

Regulation of cartilage glycosaminoglycan synthesis in the rainbow trout, Oncorhynchus mykiss, by 3,3',5-tri-iodo-L-thyronine and IGF-I.

The actions of 3,3',5-tri-iodo-L-thyronine (T3) and recombinant human IGF-I (rhIGF-I) as well as their interaction on cartilage growth in rainbow trout (Oncorhynchus mykiss) were examined. Uptake of 3H-methyl thymidine and 35S-sulfate by isolated branchial cartilage was measured as a marker for chondrocyte proliferation and sulfated glycosaminoglycan synthesis respectively. When T3 (1.0 microgram/g) was injected intraperitoneally, plasma T3 levels reached a transient peak after 1 day and decreased rapidly thereafter. Sulfate and thymidine uptake were not affected by T3 at 1 and 3 days post-injection, but at 6 days post-injection both were significantly higher in T3-injected fish than those in controls. The stimulatory effects of a T3 injection on sulfate and thymidine uptake were dose-dependent over the range of 0.01, 0.1 and 1.0 micrograms/g. In vitro exposure of cartilage to T3 (0.075, 0.75, 7.5, 75 and 750 nM) for 6 days resulted in dose-dependent stimulation of sulfate uptake, with a maximum response at 7.5 nM and higher. T3 exposure (7.5 nM) for 2 or 3 days also increased sulfate uptake, but only slightly. Thymidine uptake was not clearly affected by T3. In vitro addition of rhIGF-I (0.075, 0.75 and 7.5 nM) increased sulfate uptake, but not thymidine uptake, dose-dependently. Compared with T3, rhIGF-I induced a greater maximum level of sulfate uptake: at 7.5 nM rhIGF-I increased the uptake 17-fold whereas T3 increased the uptake fourfold. When T3 (0.075, 0.75 or 7.5 nM) and rhIGF-I (0.1 or 1.0 nM) were added together, stimulative actions of T3 on sulfate uptake were largely additive to those of rhIGF-I. The results indicate that T3 as well as IGF-I are important modulators of sulfated glycosaminoglycan synthesis in rainbow trout cartilage.

Dominance, nutritional state, and growth hormone levels in rainbow trout (Oncorhynchus mykiss).

This study addressed three questions concerning interactions between physiology and dominance in juvenile rainbow trout: (1) the validity of a model predicting a time-dependent effect of fasting on competitive ability (i.e., the ability to obtain contested food items) was tested in a series of dominance trials between fed and progressively more fasted trout, as was (2) the association between fasting and plasma growth hormone levels. (3) The relationship between plasma growth hormone levels and the competitive ability of individual trout was also studied. The main results were as follows: (1) The predictions of the time-dependent model were supported by the fasting-dominance experiment. After 3 days, fasted fish were dominant over fed fish, whereas after 6 and 9 days, the competitive ability of fed and fasted fish was similar. After 12 days, there was a tendency for fed fish to be dominant over their fasted competitors. (2) Sampling of plasma from fed and fasted trout, after 3, 6, 9, and 12 days, demonstrated that plasma growth hormone levels increases in food-deprived rainbow trout after more than 6 days of fasting, which is consistent with previous work. (3) No difference in plasma growth hormone levels was found between paired dominant and subordinate trout. Possible interactions between nutritional state, growth hormone levels, and dominance, and their implications are discussed.

Physiological concentrations of 24,25-dihydroxyvitamin D3 rapidly decrease the in vitro intestinal calcium uptake in the Atlantic cod, Gadus morhua.

The effects of physiological concentrations of 1,25(OH)2D3 and 24,25(OH)2D3 on the intestinal Ca2+ uptake in the Atlantic cod (Gadus morhua) were investigated. The intestine was perfused, in vitro, both vascularly and through the intestinal lumen, and the Ca2+ influx was measured using 45Ca. At 2.02, 4.04, 10.1, and 20.2 nM 24,25(OH)2D3 decreased the Ca2+ influx across the intestinal mucosa by 21, 31, 34, and 28%, respectively, whereas perfusion with 1,25(OH)2D3 (concentrations: 85.3, 128, 256, and 640 pM) had no effect. The actions of 24,25(OH)2D3 were dose-related, observed within 10 to 25 min, and are concluded to affect intestinal Ca2+ influx via a rapid nongenomic mechanisms; 1,25(OH)2D3 seems to be without transcaltachic effect. Thus, for the rapid, short-term regulation of intestinal calcium uptake in the Atlantic cod, 24,25(OH)2D3 is a more important vitamin D3 metabolite, in contrast to other vertebrates in which 1,25(OH)2D3 is of major importance.

Photoperiod regulation of plasma growth hormone levels during parr-smolt transformation of Atlantic salmon: implications for hypoosmoregulatory ability and growth.

The influence of photoperiod on plasma growth hormone (GH) levels during parr-smolt transformation was studied in Atlantic salmon, using a specific salmon GH radioimmunoassay; in addition, the impact of endogenous changes in GH levels on hypoosmoregulatory ability and growth was assessed using seawater challenge tests and measurements of growth rates. Groups of fish were kept on simulated natural photoperiod or continuous light, and subgroups were subjected to reciprocal transfers between these two light regimes at different times. For Atlantic salmon on simulated natural photoperiod, GH levels increased during the parr-smolt transformation, from 0.6 ng ml-1 in January-March to 6.6 ng ml-1 in June, while the corresponding change in fish on continuous light was from 0.4 to 1.0 ng ml-1. The study demonstrates that photoperiod is a major zeitgeber for the increased GH levels during the parr-smolt transformation of Atlantic salmon. The data further support the view that exposure to continuous light into fall and winter causes a "free running" of an endogenous rhythm governing smolting and a subsequent phase-delay of the parr-smolt transformation-related increase in plasma GH levels. A strong positive nonlinear correlation was found between specific growth rate and GH levels, with growth rate increasing rapidly with increasing GH levels up to 2-3 ng ml-1 at which point near-maximal growth rate is reached, revealing that relatively small increases in GH levels may be of great importance for an increased specific growth rate.(ABSTRACT TRUNCATED AT 250 WORDS)

Tartrate resistant acid phosphatase as a marker for scale resorption in rainbow trout, Oncorhynchus mykiss: effects of estradiol-17ß treatment and refeeding.

In teleosts, a considerable part of the body calcium is found in the scales. Associated with the scales are osteoblasts and osteoclasts, and during periods of high calcium demand such as during sexual maturation or starvation, the scales can be resorbed and thereby act as an internal calcium reservoir. In mammalian bone tissue, the activity of an acid phosphatase (ACP) isoenzyme, tartrate resistant acid phosphatase (TRACP), can be used as a marker for osteoclastic activity. In the present study, an evaluation of TRACP as a marker for osteoclastic activity in teleost scales has been performed. ACP and TRACP was histologically localized at resorption sites around the edge of the scales as well as at resorption holes in the scales. The optimal conditions for biochemical measurements of ACP and TRACP activity were found to be pH 5.3, 10 mM paranitrophenylphosphate, incubated for 30 min at room temperature, and 10 mM tartrate added when required. Using TRACP as a marker, estradiol-17ß (E2) was found to increase the proportion of scales being resorbed, as well as the number and size of resorption sites per scale. Also, the scales of E2-treated fish showed weaker staining for calcium. Together, the obtained data indicate that estradiol-17ß induces osteoclastic activity in teleost scales, resulting in increased resorption of the scales. A period of refeeding following a period of starvation did not have detectable effects on the scale osteoclastic activity and scale resorption.

The interrelation between photoperiod, growth hormone, and sexual maturation of adult Atlantic salmon (Salmo salar).

The plasma profiles of growth hormone (GH) in adult male and female Atlantic salmon were determined in relation to manipulation of the photoperiod and to the development and timing of sexual maturation. Fish were exposed to natural light (NL) or NL + 24L:0D additional light over the netpens from January (ALJ) or March (ALM) to July. Thereafter, these groups were brought indoors, subdivided, and subjected to simulated natural photoperiod (SNP), continuous light (24L), or short day (8L). Assay of salmon GH by RIA in monthly plasma samples revealed that GH levels were generally < 1 ng ml-1 during January to June and were only slightly affected by additional light in January or March. ALJ-24L treatment, and to a lesser extent, ALM-24L treatment, was effective in preventing sexual maturation, and GH levels of immature fish continued to be < or = 1.5 ng ml-1. On the other hand, in sexually maturing fish, GH levels increased to 2-5 ng ml-1 months prior to ovulation. Short-day photoperiod (8L) from July advanced ovulation and spermiation, whereas continuous light from July delayed these processes. The timing of the increase of GH levels was shifted in a parallel manner, indicating a functional relationship between plasma GH levels and the process of sexual maturation.

Estradiol-17ß-induced calcium uptake and resorption in juvenile rainbow trout, Oncorhynchus mykiss.

Juvenile rainbow trout, Oncorhynchus mykiss, were injected with estradiol-17ß (E2) in order to study the source of extra calcium needed during vitellogenesis. E2-treatment increased the calcium uptake from the external medium as well as calcium mobilization from muscle and scale. Judged by the increase in plasma protein-bound calcium levels, the E2-induced increase in calcium uptake is an apparent over-mobilization of calcium, i.e., the calcium uptake of the fish is in excess of what is found bound to plasma proteins. As the calcium excretion and calcium space (calculated from free plasma calcium levels) were unaffected, the excess calcium is suggested to be incorporated into internal calcium stores. This implies that the systems regulating vitellogenesis and calcium balance are integrated on the mechanistic or endocrine level, and that E2 causes calcium mobilization of a magnitude geared to the needs of the sexually maturing female.

1,25(OH)2 vitamin D3 increases ionized plasma calcium concentrations in the immature Atlantic cod Gadus morhua.

Intraperitoneal injections of 1,25(OH)2 vitamin D3 (1,25(OH)2D3) produced hypercalcemia in the marine teleost the Atlantic cod (Gadus morhua) with increased plasma concentrations of ionized calcium (CaI) while the total plasma calcium concentrations (CaT) were unaffected. A single injection of 10 micrograms kg bw-1 of 1,25(OH)2D3 increased CaI concentrations from 1.74 +/- 0.03 mM to 1.8 +/- 0.01 mM after 24 hr in Experiment 1 and from 1.67 +/- 0.03 mM to 1.82 +/- 0.06 mM after 72 hr in Experiment 2. This hypercalcemic effect was sustained by daily injections for 5 but not 7 days. Daily injections of a lower dose of 1,25(OH)2D3, 1 microgram kg bw-1, caused hypercalcemia after 5 days (CaI increased from 1.68 +/- 0.01 mM to 1.76 +/- 0.02 mM). Plasma calcium concentrations were not affected by any of the other seco-steroids (vitamin D3, (25(OH)), vitamin D3 (25(OH)D3), or 24,25(OH)2 vitamin D3 (24,25(OH)2D3), except for a decrease in CaT concentrations after 7 daily in injections of 25(OH)D3 (2 micrograms kg bw-1). Plasma phosphate concentrations were not changed by any of the seco-steroids. It is concluded that 1,25(OH)2D3 is hypercalcemic in the marine Atlantic cod.

Absence of direct regulation of prolactin cells by estradiol-17 beta in rainbow trout (Oncorhynchus mykiss).

The effects of estradiol-17 beta (E2) implants on plasma prolactin (PRL) concentrations, pituitary PRL content and pituitary PRL mRNA levels were examined in rainbow trout (Oncorhynchus mykiss). Intact immature fish treated with 1 mg estradiol-17 beta did not show significant changes in both PRL mRNA levels and pituitary PRL content after 3 days of treatment. In a similar experiment, no changes were observed in plasma PRL levels followed during 7 days. Similarly, lack of estradiol-17 beta effect on plasma PRL levels and on final PRL pituitary content was observed in ovariectomized female rainbow trout treated during 48 days with 25 mg estradiol-17 beta and in mature male fish over a 3-week treatment period. Localization of estradiol receptor (ER) mRNAs in the pituitary was carried out by Northern blot analysis using a full-length rainbow trout estrogen receptor (rtER) cDNA as a probe. The rostral pars distalis of the pituitary which contained mostly PRL cells showed the lower amount of rtER mRNA when compared to other parts of the pituitary. Moreover, two mRNAs of different size (3.5 and 1.4 kb) were detected in different parts of the pituitary. Further hybridization experiments using probes containing part of the rtER cDNA (E domain or C and D domains) indicated that the small-sized mRNA (1.4 kb) probably encodes a truncated ER protein lacking hormone binding domain or an ER-related protein. Thus, only the 3.56 kb mRNA appeared to be involved in the regulation of pituitary function by estradiol. In situ hybridization analysis allowed a more precise localization of this rtER mRNA in the pituitary.(ABSTRACT TRUNCATED AT 250 WORDS)

1,25-Dihydroxyvitamin D3 in the Atlantic cod: plasma levels, a plasma binding component, and organ distribution of a high affinity receptor.

Physiological studies of the Atlantic cod, Gadus morhua, have suggested a role for the vitamin D3 system in this marine teleost similar to that in other vertebrates. Accordingly, the present study was carried out to assess the plasma concentrations of vitamin D3, 25-hydroxyvitamin D3 (25OHD3), and 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] in this fish. Additionally, the presence of binding proteins in plasma and target-specific tissue receptors for these vitamin D3 metabolites was studied in organs normally associated with calcium regulation. Plasma levels of 25-OHD3 (undetectable to 148 pg/ml; n = 5) were comparatively low (20-30 ng/ml), whereas the levels of vitamin D3 (approximately 30 ng/ml) and 1,25-(OH)2D3 (approximately 50 pg/ml) were comparable to levels reported in higher vertebrates. Cod plasma contained a protein that bound both 25OHD3 and 1,25-(OH)2D3. This plasma binding protein revealed low affinity for 25OHD3, did not bind G-actin, and had a sedimentation coefficient of 3.4S. High affinity 1,25-(OH)2D3 receptors [Kd, 1.02 +/- 0.36 (n = 6), 1.02 +/- 0.3 (n = 5), and 0.95 +/- 0.51 (n = 5) nM; mean +/- SEM] were found in high salt cytosols from intestine, liver, and gills, respectively, and had sedimentation coefficients (3.6-3.8S in 0.3 M KCl sucrose gradients) similar to those in higher vertebrates. No specific 1,25-(OH)2D3 binding was found in kidney, ultimobranchial glands, corpuscles of Stannius, or bone. The finding of significant quantities of 1,25-(OH)2D3 in the plasma, the presence of plasma binding proteins that bind this seco-steroid, and the localization of specific high affinity receptors for this metabolite in calcium regulatory tissues in teleosts are all consistent with a physiological role for the vitamin D3 system in the calcium regulation of the cod.