Graham Young

Smoltification and seawater adaptation in coho salmon (Oncorhynchus kisutch): plasma prolactin, growth hormone, thyroid hormones, and cortisol.

The status of circulating growth hormone and prolactin during the parr-smolt transformation and during seawater adaptation of coho salmon (Oncorhynchus kisutch) was investigated in relation to changes in plasma levels of thyroxine, triiodothyronine, and cortisol, and in hypoosmoregulatory ability. Sampling (biweekly or monthly) occurred between early February and October. When peak hypoosmoregulatory ability was achieved (mid-April), one group of fish was acclimated to seawater over a period of 18 hr and was sampled 1, 3, and 7 days after the introduction of fish to seawater and biweekly thereafter. Plasma prolactin levels rose steadily from the first sampling date to a peak of 15 ng/ml in early April, declined rapidly, and remained low until June when a second increase occurred. Prolactin declined to 2 ng/ml within 1 day of the beginning of seawater adaptation. Growth hormone increased twofold from February to late March, and achieved plateau levels of 20 ng/ml in the period from mid-April to July and then gradually declined to 10 ng/ml in September and October. Plasma levels of growth hormone in seawater-acclimated fish were similar to those of freshwater coho, but with larger fluctuations; no increase was apparent during the first week of seawater acclimation. Plasma cortisol and plasma triiodothyronine increased at the same time as plasma growth hormone; increases in plasma thyroxine occurred later. In general, both growth hormone and cortisol levels were elevated when hypoosmoregulatory ability was high. Conversely, prolactin levels generally showed a negative relationship with hypoosmoregulatory ability.

Cortisol secretion in vitro by the interrenal of coho salmon (Oncorhynchus kisutch) during smoltification relationship with plasma thyroxine and plasma cortisol

An in vitro system for the incubation of interrenal tissue (head kidney fragments) from coho salmon, Oncorhynchus kisutch, was developed in order to examine changes in interrenal sensitivity to ACTH1-24 during smoltification, using cortisol secretion as the endpoint. Time-course studies indicated that maximal cortisol accumulation in incubation media was achieved after 3 hr exposure to ACTH. There was no correlation between head kidney weight, body weight, or sex and the response of the interrenal to ACTH1-24 in vitro. Approximately monthly or bi-weekly experiments were performed during the smoltification period (February-July): tissue was preincubated in hormone-free media for 3 hr, washed twice, and then challenged with 5 X 10(-10)-5 X 10(-7) M (1.5-1500 ng/ml) ACTH1-24 for 3 hr. The pattern of cortisol secretion was similar in February, early March, and late March in the dose range of 5 X 10(-10)-5 X 10(-8) M ACTH1-24. A marked, significant increase in sensitivity to ACTH and in the steroidogenic capacity of the tissue occurred in April, but by May the response was similar to that in the pre-April period. Enhanced sensitivity and steroidogenic capacity were found in interrenal tissue taken from coho salmon in June and July. Maximal in vitro responsiveness of interrenal tissue to ACTH in April was correlated with peak plasma thyroxine titers and enhanced hypoosmoregulatory ability, but not with peak plasma cortisol titers, which occurred in May.

Smoltification and seawater adaptation in Atlantic salmon (Salmo salar): plasma prolactin, growth hormone, and thyroid hormones.

To obtain more information on the role of prolactin and growth hormone during the parr-smolt transformation of Atlantic salmon, a population of fish in fresh water was sampled from January to June during two consecutive years. Gill Na+,K+-ATPase activity increased steadily during smoltification and a plasma thyroxine peak was observed 2-3 weeks before the gill Na+,K+-ATPase peak. On the basis of these two parameters, smoltification was considered complete in our populations in April 1985 and May 1986. Two peaks in plasma growth hormone levels occurred in 1986, one in mid-April and the second in mid-May. In both cases, these peaks coincided with a peak in plasma triiodothyronine and preceded the thyroxine peak by 1-2 weeks. Moreover, the second peak which lasted for 1 month coincided with maximal gill Na+,K+-ATPase activity. A decrease in plasma prolactin levels was observed during smoltification of Atlantic salmon in 2 consecutive years. During this period of decreasing and low plasma prolactin levels, gill Na+,K+-ATPase activity increased to its highest values. Atlantic salmon smolts were also directly transferred into seawater. After 2 days or more in seawater, plasma prolactin levels were not significantly different from those on Day 0, whereas in fresh water they showed large fluctuations. All these data indicate that growth hormone may play an important role in the development of hypoosmoregulatory activity. Increased hypoosmoregulatory ability also appears to be associated with low prolactin levels.

Stimulation of coho salmon growth by insulin-like growth factor I ☆

The effect of insulin-like growth factor I on growth rate of coho salmon (Oncorhynchus kisutch) was examined. Juvenile coho salmon received implants of osmotic minipumps containing recombinant bovine insulin-like growth factor I (rbIGF-I) or saline for a period of 3 to 4 weeks. High doses of rbIGF-I (greater than 0.13 microgram.g-1.d-1) resulted in hypoglycemia and death. In 2-year-old coho salmon, 0.09 microgram.g-1.d-1 rbIGF-I administered for 25 days doubled linear growth rate and increased growth rate in weight by 40%. In rapidly growing, 1-year-old coho salmon, growth rate was not altered by rbIGF-I at 0.01 or 0.05 micrograms.g-1.d-1 for 31 days. In ration-limited fish exhibiting slow growth in the control group, rbIGF-I (0.02 microgram.g-1.d-1) increased linear growth rate by up to threefold and growth rate in weight by up to fourfold. The results indicate that exogenous treatment with mammalian IGF-I can stimulate coho salmon growth under some conditions, and that endogenous IGF-I may be an important factor in regulating growth of teleosts.

Regulation of hepatic growth hormone receptors in coho salmon (Oncorhynchus kisutch).

Factors potentially regulating hepatic growth hormone (GH) receptors in coho salmon (Oncorhynchus kisutch) have been investigated. From December to June of the first year, relative changes in hepatic 125I-sGH binding and 35SO4 incorporation by ceratobranchial cartilage were similar. Stunted salmon, which in seawater have elevated plasma GH yet fail to grow, showed lower hepatic 125I-sGH binding than did normally growing seawater salmon. However, MgCl2 treatment of stunts membranes to reveal total specific binding of 125I-sGH indicated receptor occupation by endogenous sGH. Total specific 125I-sGH binding was low in seawater stunts and remained low if these fish remained unfed after return to fresh water, but increased approximately twofold upon feeding. Total specific binding in fasted salmon in fresh water showed a trend toward decreased levels by 1 week; by 3 weeks, binding was 40% lower than in fed fish. There was a positive correlation (r = 0.600) between condition factor and total specific binding in fed and fasted salmon in fresh water. Two weeks after hypophysectomy total specific binding was 50% lower than in sham-operated control salmon, indicating pituitary regulation of GH receptors. GH treatment reduced both free and total 125I-sGH binding in salmon examined 24 hr after treatment. Treatment with recombinant bovine insulin-like growth factor I, thyroxine, or cortisol did not affect free 125I-sGH binding. Both the pituitary and nutrition appear to be prime regulators of hepatic GH receptors in coho salmon.

Enhanced response of the interrenal of coho salmon (Oncorhynchus kisutch) to ACTH after growth hormone treatment in vivo and in vitro

The effects of ovine growth hormone (GH) on the response of the interrenal of yearling coho salmon (Oncorhynchus kisutch) to ACTH were examined in two series of experiments. In the first series, GH was administered intraperitoneally (2 micrograms/g body wt, three daily injections) to coho salmon which had not undergone the parr-smolt transformation. Animals were sacrificed 30 hr after the final injection and head kidney tissue was prepared for in vitro incubation. GH had no effect on plasma cortisol levels but tissue taken from treated fish had an enhanced sensitivity to ACTH in vitro and its steroidogenic capacity was increased. In the second series of experiments, interrenal tissue was preincubated with GH in the dose range of 10-1000 ng/ml and was then challenged with ACTH or was incubated with 1-10 microM pregnenolone. GH-pretreated tissue displayed an enhanced response to ACTH. Cortisol production by tissue pretreated with 500 ng/ml GH was significantly enhanced after incubation with pregnenolone. No significant effect of GH was found after simultaneous exposure of interrenal tissue to combinations of ACTH and GH for 3 hr. Pretreatment of interrenal tissue with salmon prolactin had no significant effect on the subsequent response of tissue to ACTH. These results indicate that GH regulates interrenal activity in coho salmon and that one probable site of action is on the biosynthetic pathway between pregnenolone and cortisol.

Physiology of seawater acclimation in the striped bass, Morone saxatilis (Walbaum).

Several experiments were performed to investigate the physiology of seawater acclimation in the striped bass, Morone saxatilis. Transfer of fish from fresh water (FW) to seawater (SW; 31–32 ppt) induced only a minimal disturbance of osmotic homeostasis. Ambient salinity did not affect plasma thyroxine, but plasma cortisol remained elevated for 24h after SW transfer. Gill and opercular membrane chloride cell density and Na+,K+-ATPase activity were relatively high and unaffected by salinity. Average chloride cell size, however, was slightly increased (16%) in SW-acclimated fish. Gill succinate dehydrogenase activity was higher in SW-acclimated fish than in FW fish. Kidney Na+, K+-ATPase activity was slightly lower (16%) in SW fish than in FW fish. Posterior intestinal Na+,K+-ATPase activity and water transport capacity (Jv) did not change upon SW transfer, whereas middle intestinal Na+,K+-ATPase activity increased 35% after transfer and was correlated with an increase in Jv (110%). As salinity induced only minor changes in the osmoregulatory organs examined, it is proposed that the intrinsic euryhalinity of the striped bass may be related to a high degree of “preparedness” for hypoosmoregulation that is uncommon among teleosts studied to data.

Smoltification and Seawater Adaptation in Coho Salmon (Oncorhynchus kisutch): Plasma Calcium Regulation, Osmoregulation, and Calcitonin

In order to examine the dynamics of ion regulation, osmoregulation, and plasma calcitonin during the parr-smolt transformation (smoltification), blood and gill tissue were collected from yearling coho salmon, Oncorhynchus kisutch, from February to October. Fish were kept in fresh water (FW) throughout this period. In addition, fish were exposed to seawater (SW) at the peak of smoltification in mid-April, and samples from these fish were collected until July. Plasma osmolality, gill Na+,K+-ATPase activity, plasma levels of calcitonin, and free and total calcium and magnesium were measured. SW adaptability of FW fish was assessed throughout the study by measurements of plasma osmolality following a 24-hr exposure to seawater. The greatest hypoosmoregulatory ability occurred in April-May, although SW-adapted fish had higher plasma osmolality than FW-adapted fish at all times. Gill Na+,K+-ATPase activity in FW-adapted fish increased from April to June and increased rapidly following exposure of fish to SW, and remained elevated in SW-adapted fish. Free plasma calcium and magnesium levels increased following SW exposure, but returned to prior levels within 1 week. Netting and confinement stress during sampling caused an increase in plasma osmolality and free calcium and magnesium levels in both FW- and SW-adapted fish. Changes in hypoosmoregulatory ability during smoltification and SW adaptation were correlated with changes in gill Na+,K+-ATPase activity. A sharp transitory peak in plasma calcitonin levels occurred early in smoltification (March) and in SW-adapted fish in June. Plasma calcitonin levels gradually increased in FW-adapted fish during the period of desmoltification. However, no change in plasma calcitonin levels occurred during SW-induced hypercalcemia, suggesting that the hormone does not play a major role in short-term plasma calcium regulation in coho salmon.

Somatostatin-like immunoreactivity in the pituitary and brain of three teleost fish species: somatostatin as a potential regulator of prolactin cell function.

Somatostatin-like immunofluorescence occurs in the hypothalamus and neurohypophysis of three euryhaline teleosts: tilapia, killifish, and mudsucker. This immunofluorescence was eliminated by incubating the primary antibody with excess somatostatin or somatostatin-28 but not with urotensin II, a partial analogue of somatostatin. In all three fishes, the strongest reaction was seen in the proximal pars distalis and parts of the pars intermedia. Strongly fluorescing processes from cells of the preoptic nucleus extend toward the pituitary. Distinct fluorescence was also associated with the neurohypophysis penetrating into the rostral pars distalis in the tilapia but not in the killifish or mudsucker. In the tilapia, an extensive network of immunofluorescent fibers and small cells were present in the anterior dorsolateral telencephalon, in addition to a moderately fluorescing group of cells anterolateral to the preoptic nucleus. A small area of diffuse fluorescence was also seen in the anterior dorsolateral midbrain tegmentum. Previous physiological studies have implicated somatostatin as a regulator of prolactin cell activity in tilapia. The present study demonstrates the route by which somatostatin may be delivered to the rostral pars distalis to inhibit prolactin secretion.

Effects of cortisol and growth hormone replacement on osmoregulation in hypophysectomized coho salmon (Oncorhynchus kisutch)

Both cortisol and GH were able to reverse partially the effects of hypophysectomy on coho salmon (Oncorhynchus kisutch) as judged by improved seawater (SW) tolerance after long-term treatment; however, neither hormone significantly restored gill Na+, Na+,K+-ATPase activity. In the first experiment, gill Na+,K+-ATPase activity remained low in hypophysectomized (Hx) coho treated with cortisol (15 micrograms/g body wt, suspended in cocoa butter) for 96 hr (48 hr in fresh water followed by 48 hr in seawater). In addition, plasma sodium levels were higher in the cortisol-treated Hx animals compared with those in untreated controls. In the second experiment, treatment with cortisol-filled Silastic capsules and implants of bovine GH (mixed with cholesterol in a ratio calculated to yield a dose of 1.5 microgram/g body wt/week) for 12 days also failed to alter gill Na+,K+-ATPase activity, but did reduce plasma sodium levels in animals transferred to SW for 48 hr. These results suggest that these hormones may be involved in the development of hypoosmoregulatory ability in coho salmon. However, the failure of either hormone to restore gill Na+,K+-ATPase activity suggests that a combination of these hormones and/or an additional hormone(s) acting in a synchronized fashion may be required for full effectiveness.