Comparison between O-arm Navigation and Conventional Fluoroscopic Guidance in Corrective Posterior Fixation for Cervical Spinal Injury.

Purpose : To compare the effectiveness of O-arm navigation with that of conventional fluoroscopic guidance in corrective posterior fixation for cervical spinal injury. Methods : This retrospective comparative study involved 11 consecutive patients who underwent corrective posterior fixation using O-arm navigation or conventional fluoroscopy for cervical spinal injury between February 2016 and May 2021. Patient-specific characteristics (age and sex), number of screws, number of pedicle screws, accuracy of pedicle screw insertion, number of vertebral bodies fixed, operating time, and length of hospital stay were analyzed using the t-test. A P-value < 0.05 was considered statistically significant. Results : Corrective posterior fixation was performed under O-arm navigation in 5 patients and under conventional fluoroscopic guidance in 6. A significantly greater number of pedicle screws was used in the O-arm group (6.4 vs 2.7, P = 0.046). According to the Neo classification for pedicle screw placement, there were no grade 2 or 3 breaches. No other items showed a significant difference between the groups (P > 0.05). Conclusion : O-arm navigation can improve the accuracy of cervical pedicle screw insertion. Its introduction could expand the indications for use of pedicle screws in posterior fixation of cervical spinal injury beyond those that are possible using conventional fluoroscopy. J. Med. Invest. 69 : 273-277, August, 2022.

Successful treatment of intractable pseudomeningocele with FXIII deficiency by surgery and FXIII replacement therapy: A case report.

Pseudomeningocele is an extradural cystic collection of cerebrospinal fluid (CSF) and is rare and typically asymptomatic. However, pseudomeningocele is sometimes associated with symptoms. Whether symptomatic pseudomeningocele is best treated conservatively or surgically remains controversial. Factor XIII (FXIII) is a blood coagulation factor that also promotes fibroblast proliferation during wound healing. Although treatment of postsurgical CSF leakage with FXIII has been reported, there have been no reports on surgical treatment and FXIII replacement therapy of pseudomeningocele with FXIII deficiency. We report a case of pseudomeningocele with FXIII deficiency that was successfully treated by surgery and FXIII replacement therapy. The patient presented with symptoms of intracranial hypotension syndrome that had started a few months after laminectomy for thoracic ossification of the ligamentum flavum 2 years earlier. Magnetic resonance imaging and delayed computed tomography myelography confirmed a diagnosis of pseudomeningocele. Epidural blood patch treatment was performed twice but did not result in improvement. Furthermore, the FXIII level decreased to 56%, so the patient was also diagnosed as having acquired FXIII deficiency. We elected to treat the patient by surgery with FXIII replacement therapy. The dural injury was repaired using an artificial dura mater patch, fibrin glue, and polyglycolic acid sheets. The FXIII level was 74%-135% during the perioperative period. The patient had a good postoperative course. Postoperative magnetic resonance images showed resolution of the pseudomeningocele. There was no recurrence during 6 months of follow-up. Perioperative FXIII replacement may be a useful treatment for pseudomeningocele with FXIII deficiency.

Changes in the function and phenotype of resident peritoneal macrophages after housing in an enriched environment.

Exposure to an enriched environment (EE) affects not only brain functions but also immune responses upon viral or bacterial infections. In this study, we examined changes in the phagocytic response and chemokine production of resident peritoneal macrophages after mice had been housed under EE conditions for 6 or 8 weeks, and then explored the possibility that EE could cause a change in the macrophage phenotype by means of flow cytometry as well as quantitative RT-PCR. The percentages of EE macrophages phagocytosing S. aureus and apoptotic neutrophils were significantly larger than those of standard environment (SE) macrophages. After coculturing with S. aureus, EE macrophages tended to produce greater amounts of chemokines such as MIP-2, KC and MCP-1 than SE ones, although the increases for MIP-2 and KC were not statistically significant. As compared with SE macrophages, EE macrophages included more CD40-positive cells (M1 marker), and expressed more mRNAs of IL-6 (M1 marker) and IRF4 (M2 marker), and less mRNA of CD38 (M1 marker), suggesting either the possibility that EE macrophages are a mixed population of M1 and M2 macrophages or the possibility that they are a unique population with a mixed M1 and M2 macrophage phenotype.

Safety screw fixation technique in a case of coracoid base fracture with acromioclavicular dislocation and coracoid base cross-sectional size data from a computed axial tomography study.

Coracoid base fracture accompanied by acromioclavicular joint dislocation with intact coracoclavicular ligaments is a rare injury. Generally, an open reduction with screw fixation is the first treatment choice, as it protects the important structures around the coracoid process. This report presents a new technique of screw fixation for coracoid base fracture and provides anatomic information on cross-sectional size of the coracoid base obtained by computed tomography (CT). An axial image of the coracoid base was visualized over the neck of the scapula, and a guidewire was inserted into this circle under fluoroscopic guidance. The wire was inserted easily into the neck of scapula across the coracoid base fracture with imaging in only 1 plane. In addition, 25 measurements of the coracoid base were made in 25 subjects on axial CT images. Average length of the long and short axes at the thinnest part of the coracoid base was 13.9 ± 2.0 mm (range 10.6-17.0) and 10.5 ± 2.2 mm (6.6-15.1), respectively. This new screw fixation technique and measurement data on the coracoid base may be beneficial for safety screw fixation of coracoid base fracture.

Navigation-Assisted Total Knee Arthroplasty for Osteoarthritis with Extra-Articular Femoral Deformity and/or Retained Hardware.

Total knee arthroplasty (TKA) for osteoarthritis (OA) patients with extra-articular deformity is still challenging because angular deformity, canal sclerosis, or the retained hardware that precludes the use of the traditional intramedullary guide. In addition, atypical bone cut for intra-articular correction leads to imbalanced soft tissue gap. Furthermore, corrective osteotomy should be considered for severe deformity or para-articular deformity cases. Recently, navigation-assisted TKA has been reported to increase the accuracy of prosthetic positioning and limb alignment. This system can calculate mechanical axis regardless of extra-articular deformity, canal sclerosis, or retained hardware. Accordingly, navigation surgery has been considered to be a powerful option especially in TKAs with extra-articular deformity cases. Here, we report 3 successful navigation-assisted TKAs for osteoarthritis with extra-articular deformities and/or retained hardware. Navigation-assisted TKA is an effective and reliable alternative for patients with extra-articular deformities.

Differential regulation of Igf1 and Igf2 mRNA levels in tilapia hepatocytes: effects of insulin and cortisol on GH sensitivity.

Igf1 and Igf2 stimulate growth and development of vertebrates. In mammals, liver-derived endocrine Igf1 mediates the growth promoting effects of GH during postnatal life, whereas Igf2 stimulates placental and fetal growth and is not regulated by GH. Insulin enhances Igf1 production by the mammalian liver directly, and by increasing hepatocyte sensitivity to GH. We examined the regulation of igf1 and igf2 mRNA levels by GH, insulin, and cortisol, and the effects of insulin and cortisol on GH sensitivity in primary cultured hepatocytes of tilapia, a cichlid teleost. GH increased mRNA levels of both igf1 and igf2 in a concentration-related and biphasic manner over the physiological range, with a greater effect on igf2 mRNA level. Insulin increased basal igf2 mRNA level, and strongly increased GH-stimulated igf2 mRNA level, but slightly reduced basal igf1 mRNA level and did not affect GH-stimulated igf1 mRNA level. Cortisol inhibited GH stimulation of igf1, but increased GH stimulation of igf2 mRNA level. The synergistic effect of insulin and GH on igf2 mRNA level was confirmed in vivo. These results indicate that insulin and cortisol differentially modulate the response of igf1 and igf2 mRNA to GH in tilapia hepatocytes, and suggest that the regulation of liver Igf2 production differs between fish and mammals. Regulation of liver Igf2 production in fish appears to be similar to regulation of liver Igf1 production in mammals.

Involvement of the cAMP messenger system and extracellular Ca(2+) during hyposmotically-induced prolactin release in the Mozambique tilapia.

In accord with its role in freshwater osmoregulation, prolactin (PRL) release from the tilapia pituitary is stimulated by small, physiologically relevant reductions in plasma osmolality, a response that is mediated by an acute influx of intracellular Ca(2+) through stretch-activated Ca(2+)channels. In the present study, the role of the calcium and cyclic AMP (cAMP) messenger system in the transduction of a response to a hyposmotic stimulus was examined using dispersed PRL cells and PRL cell membrane preparations from freshwater-acclimated tilapia. When PRL cells were treated with the phosphodiesterase (PDE) inhibitor, 3-isobutyl-1-methylxanthine (IBMX) (100µM), significant increases in cAMP levels and PRL release were observed at 1h. Exposure to reduced medium osmolality (300 mOsmolal) in the presence of IBMX further augmented PRL release. Depletion of Ca(2+) from the incubation medium blocked PRL release even in the presence of IBMX. By contrast, exposure of PRL cells to cholera toxin (CTX), an activator of adenylyl cyclase (AC), stimulated PRL release and cAMP accumulation in both the presence and absence of extracellular Ca(2+). On the other hand, treatment with the Ca(2+) ionophore A23187, which elicits a large rise in intracellular free Ca(2+), reduced cAMP accumulation. Likewise, the AC activity of a PRL cell membrane preparation was reduced as extracellular Ca(2+) concentration increased from 0.1 to 1 µM. These results indicate that: (1) the stimulation of PRL release and cAMP formation by a fall in extracellular osmolality are Ca(2+)-dependent; (2) large increases in intracellular Ca(2+) attenuate cAMP formation; (3) direct agonists of cAMP messenger system, such as cholera toxin, however, stimulate PRL release independently of the extracellular Ca(2+). These findings add to the evidence that the osmosensitive response of the tilapia PRL cell is mediated through a Ca(2+)-dependent mechanism. Nevertheless, the present findings also suggest that tilapia PRL cells have the ability to rapidly augment release PRL both via a Ca(2+)-dependent manner and via a cAMP-dependent pathway in the absence of extracellular Ca(2+).

Dynamic gene expression of GH/PRL-family hormone receptors in gill and kidney during freshwater-acclimation of Mozambique tilapia.

In teleosts, prolactin (PRL) and growth hormone (GH) act at key osmoregulatory tissues to regulate hydromineral balance. This study was aimed at characterizing patterns of expression for genes encoding receptors for the GH/PRL-family of hormones in the gill and kidney of Mozambique tilapia (Oreochromis mossambicus) during freshwater (FW)-acclimation. Transfer of seawater (SW)-acclimated tilapia to FW elicited rapid and sustained increases in plasma levels and pituitary gene expression of PRL177 and PRL188; plasma hormone and pituitary mRNA levels of GH were unchanged. In the gill, PRL receptor 1 (PRLR1) mRNA increased markedly after transfer to FW by 6h, while increases in GH receptor (GHR) mRNA were observed 48 h and 14 d after the transfer. By contrast, neither PRLR2 nor the somatolactin receptor (SLR) was responsive to FW transfer. Paralleling these endocrine responses were marked increases in branchial gene expression of a Na+/Cl- cotransporter and a Na+/H+ exchanger, indicators of FW-type mitochondrion-rich cells (MRCs), at 24 and 48 h after FW transfer, respectively. Expression of Na+/K+/2Cl- cotransporter, an indicator of SW-type MRCs, was sharply down-regulated by 6h after transfer to FW. In kidney, PRLR1, PRLR2 and SLR mRNA levels were unchanged, while GHR mRNA was up-regulated from 6h after FW transfer to all points thereafter. Collectively, these results suggest that the modulation of the gene expression for PRL and GH receptors in osmoregulatory tissues represents an important aspect of FW-acclimation of tilapia.

Gene expression of growth hormone family and glucocorticoid receptors, osmosensors, and ion transporters in the gill during seawater acclimation of Mozambique tilapia, Oreochromis mossambicus.

This study characterized endocrine and ionoregulatory responses accompanying seawater (SW) acclimation in Mozambique tilapia (Oreochromis mossambicus). Changes in plasma hormones and gene expression of hormone receptors, putative osmosensors, and ion transporters in the gill were measured. Transfer of freshwater (FW)-acclimated tilapia to SW resulted in a marked elevation in plasma osmolality and a significant rise in plasma growth hormone (GH) levels at 12 hr and 14 days after transfer. Significant reductions in plasma prolactin (PRL(177) and PRL(188)) levels also occurred in SW-transferred fish; no effect of transfer upon plasma cortisol or insulin-like growth factor I was observed. Gene expression of GH receptor increased strongly 6 hr after transfer, whereas PRL receptor was lower than controls at 12 hr. By contrast, mRNA levels of somatolactin and glucocorticoid receptors were unaffected by SW transfer. Osmotic stress transcription factor 1 mRNA levels rose significantly between 3 and 12 hr, whereas the calcium-sensing receptor was unaffected. Aquaporin-3 gene expression was strongly down-regulated during SW acclimation from 12 hr until the conclusion of the experiment. Na(+)/K(+)/2Cl(-) cotransporter gene expression increased significantly 3 hr after transfer, whereas expression of Na(+)/Cl(-) cotransporter, specific to FW-type chloride cells, declined by 6 hr into SW acclimation. The response of Na(+)/H(+) exchanger was less pronounced, but showed a similar pattern to that of the Na(+)/Cl(-) cotransporter. These results suggest that acquisition of hyposmoregulatory mechanisms in Mozambique tilapia entails the coordinated interaction of systemic hormones with local factors in the gill, including hormone receptors, ion transporters, and osmosensors.

Prolactin restores branchial mitochondrion-rich cells expressing Na+/Cl- cotransporter in hypophysectomized Mozambique tilapia.

Hypophysectomy and hormone replacement therapy were conducted to investigate the regulation of branchial mitochondrion-rich cell (MRC) recruitment and hormone receptor expression in euryhaline tilapia (Oreochromis mossambicus). Gene expression and immunolocalization of Na(+)/Cl(-) cotransporter (NCC) and Na(+)/K(+)/2Cl(-) cotransporter (NKCC) were used as markers for freshwater (FW)- and seawater (SW)-type MRCs, respectively. In FW fish, hypophysectomy resulted in a significant drop in plasma osmolality, an effect associated with a marked reduction of NCC gene expression and the disappearance of MRCs with apical-NCC immunoreactivity. In contrast, hypophysectomy in SW fish did not impact plasma osmolality, NKCC, or Na(+), K(+)-ATPase(alpha1) gene expression, or the recruitment of MRCs with basolateral-NKCC. Hypophysectomized fish in SW exhibited reduced mRNA levels of prolactin (PRL) receptor 1 and growth hormone (GH) receptor in the gill; GH receptor expression was also reduced following hypophysectomy in FW. PRL replacement therapy restored NCC gene expression and the appearance of MRCs with apical NCC in both FW and SW; there was no interaction of PRL with cortisol. In FW, cortisol modestly stimulated NKCC mRNA levels, while no effect of GH was evident. In SW, no clear effects of hormone replacement on gene expression of NKCC, Na(+), K(+)-ATPase(alpha1), or hormone receptors were detected. Taken together, the essential nature of PRL to survival of Mozambique tilapia in FW is derived, at least in part, from its ability to stimulate the recruitment of MRCs that express NCC, while recruitment of SW-type MRCs does not require pituitary mediation in this euryhaline tilapia.

Tissue-specific regulation of the growth hormone/insulin-like growth factor axis during fasting and re-feeding: Importance of muscle expression of IGF-I and IGF-II mRNA in the tilapia.

The effects of prolonged nutrient restriction (fasting) and subsequent restoration (re-feeding) on the growth hormone (GH)/insulin-like growth factor (IGF) axis were investigated in the tilapia (Oreochromis mossambicus). Mean weight and specific growth rate declined within 1 week in fasted fish, and remained lower than controls throughout 4 weeks of fasting. Plasma levels of IGF-I were lower than fed controls during 4 weeks of fasting, suggesting a significant catabolic state. Following re-feeding, fasted fish gained weight continuously, but did not attain the weight of fed controls at 8 weeks after re-feeding. Specific growth rate increased above the continuously-fed controls during the first 6 weeks of re-feeding, clearly indicating a compensatory response. Plasma IGF-I levels increased after 1 week of re-feeding and levels were not otherwise different from fed controls. Plasma GH levels were unaffected by either fasting or re-feeding. No consistent effect of fasting or re-feeding was observed on liver expression of GH receptor (GH-R), somatolactin (SL) receptor (SL-R), IGF-I or IGF-II. In contrast, muscle expression of GH-R increased markedly during 4 weeks of fasting, and then declined below control levels upon re-feeding for weeks 1 and 2. Similarly, muscle expression of SL-R increased after 4 weeks of fasting, and reduced below control levels after 1 and 2 weeks of re-feeding. On the other hand, muscle expression of IGF-I was strongly reduced throughout the fasting period, and levels recovered 2 weeks after re-feeding. Muscle expression of IGF-II was not affected by fasting, but was reduced after 1 and 2 weeks of re-feeding. These results indicate that GH/IGF axis, particularly muscle expression of GH-R, SL-R and IGF-I and -II, is sensitive to nutritional status in the tilapia.

Ionoregulatory and endocrine responses to disturbed salt and water balance in Mozambique tilapia exposed to confinement and handling stress.

This study assessed the endocrine and ionoregulatory responses by tilapia (Oreochromis mossambicus) to disturbances of hydromineral balance during confinement and handling. In fresh water (FW), confinement and handling for 0.5, 1, 2 and 6h produced elevations in plasma cortisol and glucose; a reduction in plasma osmolality was observed at 6h. Elevations in plasma prolactins (PRL(177) and PRL(188)) accompanied this fall in osmolality while no effect upon growth hormone (GH) was evident; an increase in insulin-like growth-factor I (IGF-I) occurred at 0.5h. In seawater (SW), confinement and handling increased plasma osmolality and glucose between 0.5 and 6h; no effect on plasma cortisol was seen due to variable control levels. Concurrently, both PRLs were reduced in stressed fish with only transient changes in the GH/IGF-I axis. Next, the branchial expression of Na(+)/K(+)/2Cl(-) cotransporter (NKCC) and Na(+)/Cl(-) cotransporter (NCC) was characterized following confinement and handling for 6h. In SW, NKCC mRNA levels increased in stressed fish concurrently with elevated plasma osmolality and diminished gill Na(+), K(+)-ATPase activity; NCC was unchanged in stressed fish irrespective of salinity. Taken together, PRL and NKCC participate in restoring osmotic balance during acute stress while the GH/IGF-I axis displays only modest responses.

Identification and genomic sequence of a ghrelin receptor (GHS-R)-like receptor in the Mozambique tilapia, Oreochromis mossambicus.

The growth hormone secretagogue-receptor (GHS-R) Is an endogenous receptor for the gut hormone ghrelin (GRLN). Two lsoforms of GHS-R have been Identified In several animals: functional GHS-R1a and a splice variant of unknown function, GHS-R1b. Here we report Identification of a GHS-R-like receptor (GHSR-LR) In the Mozambique tilapia, Oreochromis mossambicus. The cDNA is 1584 bp In length and encodes a 384-amino acid GHS-R1a ortholog. The amino acid sequence of tilapia GHS-R1a is 54, 60, 80 and 89% Identical to that of rat, chicken, pufferfish, and seabream GHS-R1a, respectively. Genomic PCR revealed that the tilapia GHS-R gene Is composed of two exons separated by a single intron. In addition, a GHS-R1b ortholog, which Is generated by alternative splicing of the GHS-R gene and contains part of the intron, was Identified and predicted to be a 298-amino acid protein. Functional analyses of tilapia GHS-R1a were conducted using mammalian HEK 293 and CHO cells, but the expected increase in intracellular calcium Ions by tilapia or rat GRLN was not observed. We found that the GHS-R1a ortholog Is expressed In greater quantities than the GHS-R1b ortholog in all tissues assayed. Further studies are required to conclude that our Identified protein Is the GHS-R for tilapia, although the gene structure and amino acid sequence showed high similarities to other GHS-R genes; thus, we designated this protein GHSR-LR.

Induction of vitellogenin production in male tilapia (Oreochromis mossambicus) by commercial fish diets.

Mozambique tilapia, (Oreochromis mossambicus), are a euryhaline teleost and an important biological model species. Captive male tilapia frequently have high levels of the estrogen-induced yolk precursor protein vitellogenin (Vg), a common indicator of exposure to estrogenic compounds. Sex steroids are found in commercial fish diets, but relatively few studies have examined the relationship between commercial diets and Vg production. In a fasting experiment to ascertain a dietary role in male Vg production, plasma Vg was reduced to negligible levels after 2 weeks of fasting, while no change in estrogen receptor (ER) expression was seen. When male tilapia were fed a squid-based diet that replaced the commercial trout diet, plasma Vg was reduced to undetectable levels over 40 days, concomitant with significant reductions in hepatic expression of Vgs A, B, and C, and ERbeta, compared with control fish fed commercial trout diet. Female tilapia fed the squid-based for 20 days had no change in these parameters. When male tilapia were fed a defined, soy-based diet, plasma Vg reduced to 20% of levels in fish given either commercial trout diet or a defined, fishmeal-based diet. Overall, results from these studies suggest that estrogens in a commercial trout diet induce vitellogenin production by increasing expression of Vg, but not ER genes in male tilapia.

Effects of short- and long-term fasting on plasma and stomach ghrelin, and the growth hormone/insulin-like growth factor I axis in the tilapia, Oreochromis mossambicus.

Ghrelin is a highly conserved peptide hormone secreted by the stomach, which is involved in the regulation of food intake and energy expenditure. Ghrelin stimulates growth hormone (GH) release, and increases appetite in a variety of mammalian and non-mammalian vertebrates, including several fish species. Studies were conducted to investigate the effect of feeding and fasting on plasma and stomach ghrelin, and the growth hormone/insulin-like growth factor I (IGF-I) axis in the Mozambique tilapia, a euryhaline teleost. No postprandial changes in plasma and stomach ghrelin levels or stomach ghrelin mRNA levels were observed. Plasma levels of GH, IGF-I and glucose all increased postprandially which agrees with the anabolic roles of these factors. Fasting for 4 and 8d did not affect ghrelin levels in plasma or stomach. Plasma GH was elevated significantly after 4 and 8d of fasting, while plasma IGF-I levels were reduced. Plasma ghrelin levels were elevated significantly after 2 and 4 wk of fasting, but no change was detected in stomach ghrelin mRNA levels. Four weeks of fasting did not affect plasma GH levels, although plasma IGF-I and glucose were reduced significantly, indicating that GH resistance exists during a prolonged nutrient deficit (catabolic state). These results indicate that ghrelin may not be acting as a meal-initiated signal in tilapia, although it may be acting as a long-term indicator of negative energy balance.

cDNA cloning and isolation of somatolactin in Mozambique tilapia and effects of seawater acclimation, confinement stress, and fasting on its pituitary expression.

Somatolactin (SL) is a member of the growth hormone (GH)/prolactin (PRL) family of pituitary hormones, and is found in a variety of teleost species. Somatolactin is thought to be involved in a wide range of physiological actions, including reproduction, stress response, the regulation of Ca(2+) and acid-base balance, growth, metabolism, and immune response. We report here on the cDNA structure of SL from the pituitary of Mozambique tilapia, Oreochromis mossambicus, and its gene expression in response to seawater acclimation, stress, and fasting. Tilapia SL cDNA (1573bp long) encoded a prehormone of 230 amino acids. Sequence analysis of purified SL revealed that the prehormone is composed of a signal peptide of 23 amino acids and a mature protein of 207 amino acids, which has a possible N-glycosylation site at position 121 and seven Cys residues. Tilapia SL shows over 80% amino acid identity with SLalpha of advanced teleosts such as medaka and flounder, and around 50% identity with SLbeta of carp and goldfish. Acclimation to seawater had no effect on pituitary expression of SL or on hepatic expression of the putative tilapia SL receptor (GHR1). By contrast, seawater acclimation resulted in significant increases in pituitary GH expression and in hepatic expression of tilapia GH receptor (GHR2). Confinement stress had no effect on pituitary expression of either SL or GH, or on hepatic expression of GHR1, whereas a significant increase was seen in GHR2 expression in the liver. Fasting for 4 weeks resulted in significant reductions in SL transcripts both in fresh water and seawater. It is highly likely that SL is involved in metabolic processes in tilapia along with the GH/IGF-I axis.

Effects of o,p'-DDE, heptachlor, and 17beta-estradiol on vitellogenin gene expression and the growth hormone/insulin-like growth factor-I axis in the tilapia, Oreochromis mossambicus.

Effects of two endocrine disruptors, o,p'-DDE and heptachlor, and 17beta-estradiol (E(2)) on vitellogenin (Vg) and the growth hormone (GH)/insulin-like growth factor-I (IGF-I) axis were examined in male tilapia. In the first experiment, fish were given 5 weekly injections of either E(2), o,p'-DDE or heptachlor (5 microg/g). E(2) treatment increased plasma Vg and hepatic expression of three Vg genes (Vgs A, B, and C) and estrogen receptor alpha (ERalpha), while reducing plasma levels of IGF-I and suppressing the expression of IGF-I, the GH receptor (GHR2) and the putative somatolactin receptor (GHR1). Neither pesticide greatly affected the other parameters examined, except for a significant reduction in expression of GHR2 and increased plasma IGF-I. In the second experiment, fish were given a single injection of o,p'-DDE or heptachlor (100 microg/g), or E(2) (5 microg/g) and sacrificed 5 days post-injection. Treatment with E(2) stimulated expression of all three Vg genes. Both o,p'-DDE and heptachlor increased expression of VgB, whereas only o,p'-DDE increased VgA expression. There was no effect of o,p'-DDE or heptachlor on VgC expression or plasma Vg levels. Treatment with o,p'-DDE and heptachlor as well as E(2) increased ERalpha and ERbeta transcript levels. Similarly, both pesticides increased GHR1 and IGF-I expression, whereas no significant effect of E(2) was observed on GHR1, GHR2 or IGF-I expression. These results indicate that o,p'-DDE and heptachlor have varying temporal and dose effects on modulation of Vg and the GH/IGF-I axis that are distinct from E(2).

Osmosensitivity of prolactin cells is enhanced by the water channel aquaporin-3 in a euryhaline Mozambique tilapia (Oreochromis mossambicus).

In teleost fish, prolactin (PRL) has important actions in the regulation of salt and water balances in freshwater (FW) fish. Consistent with this role, the release of PRL from the pituitary of the Mozambique tilapia is stimulated as extracellular osmolality is reduced. Stretch-activated calcium-permeant ion channels appear to be responsible for the initiation of the signal transduction that leads to increased PRL release when PRL cells are exposed to reductions in extracellular osmolality. In this study, we examined a possible involvement of the aquaporin-3 (AQP3) water channel in this osmoreceptive mechanism in PRL cells of the tilapia. AQP3 expression levels in the rostral pars distalis of the pituitary, consisting predominantly of PRL cells, were higher in fish adapted to FW than in seawater (SW)-adapted fish. Immunohistochemical studies revealed that AQP3 is located in the cell membrane and perinuclear region of PRL cells, with more intense immunosignals in PRL cells of FW-adapted fish than in those of SW fish. In FW PRL cells, the magnitude of hyposmoticity-induced cell volume increase was greater than that seen in SW PRL cells. Mercury, a potent inhibitor of AQP3, inhibited hyposmoticity-induced cell volume increase and PRL release from FW PRL cells. The inhibitory effect of mercury was partially restored by beta-mercaptoethanol, whereas no effect of mercury was observed on PRL release stimulated by a depolarizing concentration of KCl, which induces Ca2+ influx and stimulates the subsequent Ca2+-signaling pathway. These results indicate significant contribution of AQP3 to osmoreception in PRL cells in FW-adapted tilapia.