Pseudoplatystoma Corruscans Spix Amp Agassiz 1829-Books Pdf

Pseudoplatystoma corruscans Spix amp Agassiz 1829
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238 Bile acids as potential pheromones in Pseudoplatystoma corruscans. al 2001 For instance behavioral evidence suggests that communication because such species show alarm chemicals. one of their functions as olfactory stimulants is their role as from club cells and are even able to recognize conspecific. pheromones for migratory anadromous fishes some of which size by chemical information Giaquinto Volpato 2005. appear to recognize and select the odor of conspecifics when Also chemical signals are likely to be an important source. choosing spawning streams Doving et al 1980 Stabell of information in feeding behavior Giaquinto Hoffmann. 1992 Recently it has been shown that bile acid profiles of 2010 and in female mate choice Giaquinto 2010 These. lake trout Salvelinus namaycush are largely influenced by confirm the importance of olfaction for this taxonomic group. sex and maturation stage Zhang et al 2001 and that bile of fish Also as a nocturnal and predatory fish chemically. of female Rainbow trout Oncorhynchus mykiss contains a modulated reactions are expected to play a major role in intra. pheromone Vermeirssen Scott 2001 Moreover male and inter specific relationships Based on these findings we. specific bile acids of sea lamprey Petromyzon marinus are planned to investigate a possible biological role of bile acids. potent and specific stimulants to the female olfactory organ in pintado Pseudoplatystoma corruscans by testing which. Siefkes Li 2004 Furthermore electrophysiological of them would be detected by olfaction. studies demonstrated that an odotopic map of responses Despite growing interest in the role of bile acids in. to bile acids and amino acids are represented in different fish chemoreception Zhang et al 2001 little effort has. regions of olfactory bulbs of chars Salvelinus alpinus been made to examine a possible pheromonal role of bile. and graylings Thymallus thymallys Doving et al 1980 acids in fish To investigate pheromonal roles of bile acids. Behavioral studies in Atlantic salmon Salmo salar also pintado our fist study tested which bile acids are detected by. showed that intestinal extracts which are known to contain olfaction using electro olfactogram EOG recordings From. bile acids induced strain specific preference and searching 30 bile acids tested five showed evident EOG responses. behaviors Stabell 1992 Moreover lake char Salvelinus among them taurocholic acid TCA was investigated in a. namaycush exhibit preference behaviors towards water behavioral study as a possible pheromone because it showed. containing bile acids Zhang et al 1996 Indeed several the strongest response and previous reports showing that this. experiments indicated that a number of bile acids induced bile acid is released in feces in high proportion Zhang et al. behavioral responses in fish Jones Hara 1985 Hara 2001 and authentic bile acids are also important inducing. 1994 and therefore support our hypothesis that bile acids behavioral responses in fish Jones Hara 1985. can act as pheromones, In teleosts the main bile acids are Hara 1994 sulphated Material and Methods. bile alcohol mainly 5 cyprinol and 5 chimaerol and. Zhang et al 2001 C24 bile acids mainly cholic acid CA Animals and housing Pintado fish Pseudoplatystoma. chenodeoxycholic acid CD deoxycholic acid DC and corruscans were obtained from a commercial fish farm. haemulcholic acid Haslewwod 1967 Goto et al 1996 The and kept in indoor 500 L tanks for 30 days prior to the. C24 bile acids are taurine conjugated and or sulphated Also experimental procedures Water temperature ranged between. cysteinolic acid conjugated bile acids were found in the bile 28 30 C and a 12L 12D photoperiod was maintained. of some marine species Une et al 1991 Goto et al 1996 The tanks were constantly aerated and ammonium and. The bile acid composition of rainbow trout was investigated nitrite levels were maintained below 0 5 and 0 05 mg L. and cholic acid was found to be the main component and respectively The tanks were provided with PVC tubes as. constituted over 85 of total Denton Yousef 1974 shelters for the fish The fish were daily fed with two live. Chenodeoxycholic acid accounted for 14 and 3 12 7 keto worms per fish. and 7 12 3 keto 5 cholanoates for 1 or less of total In The length and mean weight of these fish was 9 6 0 92. lake char taurocholic acid TCA and Taurochenodeoxycholic cm and 31 2 0 58 g respectively Fish were not separated. acid TCD are the two principal bile acids Zhang et al by sex since they are immature at this size Godinho et al. 2001 and these results are consistent with previous reports 2007. in other salmonid species Sasaki 1966 Haslewood 1967. Denton Yousef 1974 In spite that all these substances Experimental design We tested several types of bile. are well known no specific bile acids have been identified as acids to analyze their potential as chemical signal Thus. pheromones in catfish species These fish are known to have we firstly tested the response of chemosensory apparatus. taste buds all around their body surface used for perceiving by using EOG record Secondly we chose the strongest bile. environmental chemical cues Barlow Northcutt 1997 acid to test its role in a two choice maze running water. and have also specific adaptations the barbells used for containing bile acid or a plain water as control. the same purpose Kotrschal et al 1996 Regarding, intraespecific chemical communication catfish are known Stimulus administration and EOG recording Each. to release alarm pheromone when their skin is damaged fish was tranquilized with MS222 1 8000 anaesthetized. causing conspecific antipredator responses Giaquinto intraperitoneally with amobarbital 30 mg kg body weight. Volpato 2001 The pintado Pseudoplatystoma corruscans and immobilized with an intramuscular injection of Flaxedil. is an interesting South American catfish to study chemical gallamine triethiodide 3 4 mg kg body weight The. P C Giaquinto R E Barreto G L Volpato M Fernandes de Castilho E Gon alves de Freitas 239. anaesthetized fish was wrapped with an absorbent tissue Nikonov Caprio 2001 Zhang et al 2001 Rolen et al. and secured in a holding apparatus The right side rosette 2003 Rolen Caprio 2007 commonly utilized one or. was exposed by removing the dorsal aspect of the skin and more of these bile salts in their studies. the cartilage of the olfactory sac The naris and the gills Stock solutions of the stimulants 10 3 M concentrations. were perfused with plain water To deliver plain water or were prepared with distilled water and stored in a. stimulants to the naris the method of Sveinsson 2000 was refrigerator Test solutions were diluted with plain water. used Briefly a pneumatic activator switches plain water before testing To eliminate the effect of distilled water. to from the stimulant solution delivered by two identical aliquots of stock solutions were diluted at least 100 times. polyethylene tubes The pneumatic valve was controlled by of plain water to form test formulas. a solenoid and an associated electronic timing device Hara. et al 1973 The solution flowed through a glass capillary Cross adaptation Cross adaptation developed by Caprio. positioned over the rosette 10 mL min This system Byrd 1984 was used to compare the EOG response to. provides an approximate square stimulation at required a test stimulus before and during adaptation to an adapting. concentration each time with no apparent interruption or compound using a protocol by Li Sorensen 1997 In a. disturbance of the flow to the naris The stimulus duration given trial baseline olfactory EOG responses of pintado. was 10s to blank water control control A an L arginine standard. EOG responses were recorded differentially by a pair and test stimuli bile acids were recorded The test stimuli. of Ag AgCl electrodes type MEH1S World Precision were used at concentrations that elicited approximately. Instruments FL USA filled with 3M KCl bridged by equipotent olfactory responses at about 100 of the. 8 gelatin saline filled capillaries tip diameter 100 150 L arginine standard During adaptation the olfactory. m Evans Hara 1985 One electrode was positioned epithelium was continually exposed to the adapting stimulus. near the central ridge posterior portion of the rosette and for 5 min after which a 5 s application of the same adapting. slightly above the olfactory epithelium The other electrode stimulus was tested first at the concentration used in the. reference was placed on the skin surface adjacent to adaptation control B and then at twice the concentration. the perfused olfactory cavity Electrical activities were used for the adaptation self adapted control Then the. amplified with a DC preamplifier Grass 7P1 Grass other test stimuli were tested in the adapting solution. Instruments MA USA adapted response Each test was interspersed with 5 s. applications of the L arginine standard and control B to. Chemicals In total 30 bile acids were tested which confirm the responsiveness of the tested fish Switching. comprised all known commercially available bile acids the adapting stimulus back to blank water completed the. produced by fish and common vertebrates All tested trial The epithelium of the fish was allowed to recover for. bile acids were purchased from either Sigma Chemical 30 min and then was tested again using another adapting. St Louis M O USA or Steraloids Inc Wilton NH stimulus Cross adaptation data were expressed as percent. USA The five most potent bile acids were select for initial response PIR using the following formula adapted. Concentration Response tests Names and abbreviations of from Caprio Byrd 1984 and Li Sorensen 1997. most potent bile acids are listed in Table 1 5 out 30 tested. Table 1 Trivial names for bile acids are used throughout PIR adapted response control B x 100. the manuscript initial response control A, Table 1 Names and abbreviations for the tested bile acids where a larger PIR indicates less cross reactivity between. Abbreviation Trivial name Chemical name olfactory receptor mechanisms or separate receptor sites and. a low PIR indicates more cross reactivity or shared receptor. 3 7 12 Trihydroxy 5 cholan, CA Cholic acid sites and or a common signal transduction pathway. 24 oic acid,3 7 Dihydroxy 5 cholan 24 oic,CD Chenodeoxycholic acid.
acid Statistical analysis of EOG data Statistical analysis. 3 12 Dihydroxy 5 cholan 24 oic system SAS software was used to conduct all analyses. DC Deoxycholic acid, In concentration response experiments responses were. 3 7 12 Trihydroxy 5 cholan 24,TCA Taurocholic acid. oic acid N 2 sulfoethy amide visually compared The lowest concentration at which a. Taurochenodeoxycholic 3 7 Dihydroxy 5 cholan 24 oic stimulus elicited a response larger than the blank water. acid acid N 2 sulfoethy amide control Student s t test was considered to be its response. threshold In cross adaptation experiments a paired t test. The five bile salts tested in this study were selected to was used to determine if responses to a test stimulus during. include two produced by the channel catfish TCDC and adaptation were significantly different from the responses. TCA Kellogg 1975 Previous investigators D ving to the same test stimulus before adaptation All PIR data. et al 1980 Goh Tamura 1980 Jones Hara 1985 were subsequently analyzed to directly detect cross. Hellstr m D ving 1986 Friedrich Korsching 1998 reactivity by subjecting all PIR data to a two way analysis. 240 Bile acids as potential pheromones in Pseudoplatystoma corruscans. of variance ANOVA If the main effect was found to Results. be significant the PIR data was divided into five groups. according to adapting stimulus For each group responses EOG responses Concentration response C R. of adapted epithelia to test stimuli were analyzed by a relationships of the five bile acids were plotted together. one way ANOVA to determine the affect of the adapting as percentages of the L arginine standard Five most. stimulus Again if the main effect was significant the potent bile acids were DC CD CA TCD and TCA with. significance of the adapting effect for each test stimulus detection thresholds of 0 02 nM considered extremely low. was determined by comparing all PIRs in the group with concentrations regarding chemical cues D ving et al. the PIR of the self adapted control using Dunnett s test 1980 Hara et al 1984 Hara Zhang 1996 1998 Bile. which tests for differences between several treatments acids were grouped according to the molecular structures. and a single control The following categories were used Fig 1 Responses to all concentrations of bile acids. to classify cross adaptation responses 1 not adapted ranged from 9 to 640 and the blank water control. meaning responses during adaptation were not significantly elicited a mean response of 7 9 6 5 mean SE The. different from the initial response paired t test P 0 05 free bile acids deoxycholic acid DC chenodeoxycholic. 2 partially adapted meaning responses during adaptation CD and cholic acid CA had similar nearly parallel C R. were significantly less than the initial responses paired curves with detection thresholds 10 9 M 10 11 M and 10 9 M. t test P 0 05 but the PIR were significantly greater than respectively Fig 1 Both the bile acids conjugated with. the control Dunnetts P 0 05 and 3 adapted to control taurine taurochenodeoxycholic acid TCD and taurocholic. levels meaning PIR were not significantly different than acid TCA had essentially identical C R curves with. Volpato 2001 The pintado Pseudoplatystoma corruscans is an interesting South American catfish to study chemical communication because such species show alarm chemicals from club cells and are even able to recognize conspecific size by chemical information Giaquinto amp Volpato 2005 Also chemical signals are likely to be an important source

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