ABSTRACT
This study approached the long-term oral administration of cortisol (F) and dexamethasone (DEX), two synthetic glucocorticoids, compared to a control group (CT) in the juveniles of a marine teleost, the gilthead seabream (Sparus aurata). Physiologically, DEX treatment impaired growth, which appears to be linked to carbohydrate allocation in muscle and liver, hepatic triglycerides depletion, and reduced hematocrit. Hypophyseal gh mRNA expression was 2-fold higher in DEX than in CT or F groups. Similarly, hypothalamic trh and hypophyseal pomcb followed this pattern. Plasma cortisol levels were significantly lower in DEX than in CT, while F presented intermediate levels. In the posterior intestine, measured short circuit-current (Isc) was more anion absorptive in CT and F compared to the DEX group, whereas Isc remained unaffected in the anterior intestine. The derived transepithelial electric resistance (TEER) significantly differed between intestinal regions in the DEX group. These results provide new insights to understand better potential targeted biomarkers indicative of the differential glucocorticoid or mineralocorticoid-receptors activation in fish.
Subject(s)
Sea Bream , Animals , Sea Bream/metabolism , Hydrocortisone/metabolism , Intestines , Hypothalamus , Glucocorticoids/metabolism , Dexamethasone/pharmacology , Dexamethasone/metabolismABSTRACT
Two orthologues of the gene encoding the Na+-Cl- cotransporter (NCC), termed ncca and nccb, were found in the sea lamprey genome. No gene encoding the Na+-K+-2Cl- cotransporter 2 (nkcc2) was identified. In a phylogenetic comparison among other vertebrate NCC and NKCC sequences, the sea lamprey NCCs occupied basal positions within the NCC clades. In freshwater, ncca mRNA was found only in the gill and nccb only in the intestine, whereas both were found in the kidney. Intestinal nccb mRNA levels increased during late metamorphosis coincident with salinity tolerance. Acclimation to seawater increased nccb mRNA levels in the intestine and kidney. Electrophysiological analysis of intestinal tissue ex vivo showed this tissue was anion absorptive. After seawater acclimation, the proximal intestine became less anion absorptive, whereas the distal intestine remained unchanged. Luminal application of indapamide (an NCC inhibitor) resulted in 73% and 30% inhibition of short-circuit current (Isc) in the proximal and distal intestine, respectively. Luminal application of bumetanide (an NKCC inhibitor) did not affect intestinal Isc. Indapamide also inhibited intestinal water absorption. Our results indicate that NCCb is likely the key ion cotransport protein for ion uptake by the lamprey intestine that facilitates water absorption in seawater. As such, the preparatory increases in intestinal nccb mRNA levels during metamorphosis of sea lamprey are likely critical to development of whole animal salinity tolerance.
Subject(s)
Ion Transport/genetics , Osmoregulation/genetics , Petromyzon/genetics , Salt Tolerance/genetics , Solute Carrier Family 12, Member 3/genetics , Amino Acid Sequence , Animals , Bumetanide/pharmacology , Fresh Water/chemistry , Gills/metabolism , Indapamide/pharmacology , Intestines/metabolism , Ion Transport/drug effects , Metamorphosis, Biological/drug effects , Metamorphosis, Biological/genetics , Petromyzon/metabolism , Phylogeny , RNA, Messenger/genetics , Real-Time Polymerase Chain Reaction/methods , Salinity , Salt Tolerance/drug effects , Seawater/chemistry , Sodium Chloride Symporter Inhibitors/pharmacology , Sodium Potassium Chloride Symporter Inhibitors/pharmacology , Sodium-Potassium-Chloride Symporters/genetics , Sodium-Potassium-Chloride Symporters/metabolism , Solute Carrier Family 12, Member 3/metabolism , Water/metabolismABSTRACT
Lampreys are the most basal vertebrates with an osmoregulatory strategy. Previous research has established that the salinity tolerance of sea lamprey increases dramatically during metamorphosis, but underlying changes in the gut have not been examined. In the present work, we examined changes in intestinal function during metamorphosis and seawater exposure of sea lamprey (Petromyzon marinus). Fully metamorphosed juvenile sea lamprey had 100% survival after direct exposure to 35 parts per thousand seawater (SW) and only slight elevations in plasma chloride (Cl-) levels. Drinking rates of sea lamprey juveniles in seawater were 26-fold higher than juveniles in freshwater (FW). Na+-K+-ATPase (NKA) activity in the anterior and posterior intestine increased 12- and 3-fold, respectively, during metamorphosis, whereas esophageal NKA activity was lower than in the intestine and did not change with development. Acclimation to SW significantly enhanced NKA activity in the posterior intestine but did not significantly change NKA activity in the anterior intestine, which remained higher than that in the posterior intestine. Intestinal Cl- and water uptake, which were observed in ex vivo preparations of anterior and posterior intestine under both symmetric and asymmetric conditions, were higher in juveniles than in larvae and were similar in magnitude of those of teleost fish. Inhibition of NKA by ouabain in ex vivo preparations inhibited intestinal water absorption by 64%. Our results indicate drinking and intestinal ion and water absorption are important to osmoregulation in SW and that preparatory increases in intestinal NKA activity are important to the development of salinity tolerance that occurs during sea lamprey metamorphosis.
Subject(s)
Drinking , Intestinal Absorption , Intestines/physiology , Osmoregulation , Petromyzon/physiology , Salt Tolerance , Seawater , Animals , Chlorides/metabolism , Fish Proteins/metabolism , Intestines/enzymology , Metamorphosis, Biological , Petromyzon/metabolism , Potassium/metabolism , Salinity , Sodium/metabolism , Sodium-Potassium-Exchanging ATPase/metabolismABSTRACT
Fish are continuously forced to actively absorb or expel water and ions through epithelia. Most studies have focused on the gill due to its role in Na+ and Cl- trafficking. However, comparatively few studies have focused on the changing function of the intestine in response to external salinity. Therefore, the present study investigated the main intestinal changes of long-term acclimation of the Senegalese sole (Solea senegalensis) to 5, 15, 38 and 55ppt. Through the measurement of short-circuit current (Isc) in Ussing chambers and biochemical approaches, we described a clear anterior/posterior functional regionalization of the intestine in response to salinity. The use of specific inhibitors in Ussing chamber experiments, revealed that the bumetanide-sensitive Na+/K+/Cl- co-transporters are the main effectors of Cl- uptake in both anterior intestine and rectum. Additionally, the use of the anion exchanger specific inhibitor, DIDS, showed a salinity/region dependency of anion exchanger function. Moreover, we also described ouabain-sensitive Na+/K+-ATPase (NKA) and Bafilomycin A1-sensitive H+-ATPase activities (HA), which displayed changes related to salinity and intestinal region. However, the most striking result of the present study is the description of an omeprazole-sensitive H+/K+-ATPase (HKA) in the rectum of Senegalese sole. Its activity was consistently measurable and increased at lower salinities, reaching rates even higher than those of the NKA. Together our results provide new insights into the changing role of the intestine in response to external salinity in teleost fish. The rectal activity of HKA offers an alternative/cooperative mechanism with the HA in the final processing of intestinal water absorption by apical titration of secreted bicarbonate.
Subject(s)
Flatfishes/physiology , Intestines/physiology , Salinity , Adenosine Triphosphatases/metabolism , Animals , Gills/physiology , Intestines/enzymologyABSTRACT
OBJECTIVES: We studied the left ventricular systolic and diastolic function in preterm infants during the first month of life in comparison with the respective patterns in term neonates. STUDY DESIGN: Serial 2-dimensional/M-mode and Doppler transmitral flow velocity measurements were performed in 20 preterm (gestational age, 32 to 36 weeks) and 25 term infants between days 2 and 5 after birth and at age 1 month. RESULTS: After birth, the early velocity, early integral, early filling fraction, early and atrial velocity ratios, and early and atrial integral ratios were lower in the preterm than in the term infants (P <.01). During the first 4 weeks of life, these values, the atrial velocity, and the atrial integral increased gradually, whereas the atrial filling fraction, diastolic filling time, and deceleration time of early diastolic filling decreased slightly with postnatal age (P <.01) in the preterm infants. At age 1 month their diastolic function reached the level of term infants with the exception of early and atrial integral ratios and atrial filling fraction. The measures of systolic performance were within normal range, but midwall fractional shortening at 2 to 5 days (P <.05), and fractional shortening area (P <.05), as well as midwall fractional shortening (P <.01) at 1 month of age, were slightly inferior in the preterm than in the term infants. During the first month the left ventricular mass and left ventricular mass/body surface area ratio increased more markedly in the preterm than term infants, significantly in both groups (P <.05). CONCLUSIONS: The preterm diastolic patterns represent a transition between the patterns of the fetus and those of term neonates. These changes reflect an improvement in the left ventricular diastolic function, more markedly in relaxation than in compliance. The postnatal increase in the transmitral measures and left ventricular mass suggest marked myocardial adaptation to the extrauterine environment in the preterm infant already during the first month.
Subject(s)
Diastole/physiology , Infant, Premature/physiology , Systole/physiology , Ventricular Function, Left/physiology , Blood Flow Velocity/physiology , Child Development/physiology , Echocardiography, Doppler , Female , Follow-Up Studies , Humans , Infant, Newborn , Male , Prospective Studies , Stroke Volume/physiologyABSTRACT
Postnatal changes in left ventricular diastolic filling and systolic cardiac performance were studied monthly by serial echocardiographic measurements from days 3-5 up to six months in 20 healthy full-term infants. The fractional shortening area (FSA = (left ventricular end-diastolic area - end-systolic area)/end-diastolic area) was assessed for systolic performance, and transmitral pulsed-wave Doppler flow velocity patterns were analysed for diastolic function. FSA remained stationary during the observation. After birth, left ventricular peak early (E) and atrial (A) velocities and the respective integrals were lower than at one month of age (47+/-5 vs. 63+/-6 cm/s and 44+/-6 vs. 57+/-5 cm/s and 3.33+/-0.40 vs. 4.05+/-0.53 cm and 2.74+/-0.40 vs. 3.18+/-0.53 cm; P < 0.05). During the next five months, the early parameters (E velocity and E integral) increased but the atrial indices (A velocity and A integral) did not change. During the whole observation the E/A velocity ratio, the E/A integral ratio and the early filling fraction (EFF) increased. The early filling deceleration time was longer during the first month than later (87+/-10 vs. 72+/-13 ms; P < 0.05). In conclusion, age-related changes were observed in the diastolic but not in the systolic performance in healthy full-term infants during the first six months. The most intensive changes took place in the early and atrial transmitral parameters during the first month of life, suggesting an improvement in both left ventricular relaxation and compliance. During the following five months, the early mitral parameters increased but the atrial diastolic values remained stable. These changes may mainly be determined by the improvement in left ventricular relaxation.