University of Richmond UR Scholarship Repository Master's eses Student Research 5-1973 Changes in the (Na+ K+) - dependent ATPase activity in gills of the euryhaline teleost, Fundulus heteroclitus, upon adaptation to freshwater and salt water as determined by specific activities and 3H - ouabain binding sites Mary Elizabeth Gilman Follow this and additional works at: hp://scholarship.richmond.edu/masters-theses is esis is brought to you for free and open access by the Student Research at UR Scholarship Repository. It has been accepted for inclusion in Master's eses by an authorized administrator of UR Scholarship Repository. For more information, please contact [email protected]. Recommended Citation Gilman, Mary Elizabeth, "Changes in the (Na+ K+) - dependent ATPase activity in gills of the euryhaline teleost, Fundulus heteroclitus, upon adaptation to freshwater and salt water as determined by specific activities and 3H - ouabain binding sites" (1973). Master's eses. Paper 468.
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University of RichmondUR Scholarship Repository
Master's Theses Student Research
5-1973
Changes in the (Na+ K+) - dependent ATPaseactivity in gills of the euryhaline teleost, Fundulusheteroclitus, upon adaptation to freshwater and saltwater as determined by specific activities and 3H -ouabain binding sitesMary Elizabeth Gilman
Follow this and additional works at: http://scholarship.richmond.edu/masters-theses
This Thesis is brought to you for free and open access by the Student Research at UR Scholarship Repository. It has been accepted for inclusion inMaster's Theses by an authorized administrator of UR Scholarship Repository. For more information, please [email protected].
Recommended CitationGilman, Mary Elizabeth, "Changes in the (Na+ K+) - dependent ATPase activity in gills of the euryhaline teleost, Fundulusheteroclitus, upon adaptation to freshwater and salt water as determined by specific activities and 3H - ouabain binding sites" (1973).Master's Theses. Paper 468.
i hr. to 24 hr. ) 9 .0914 i .0093 SW (fully adaptGd) 5 .0505 + - .0041 SW Ci hr. to 36 hr.) 9 .0534 + .0054 SW (fully adapted +
i hr. to )6 hr. ) 14 .0524 + .0037 -
.FW = freshwater SW = salt water (17 ppt)
; 1.' •• :
28
Table 3. Results of T - test for significance between means of (Na++K+) - dependent ATPase in gill homogenates (2%) of ~. heteroclitus.
x y df
SW - F FW - F 8
SW - T FW - F 12 SW - F + SW - T FW - F 17 SW - F FW - F + FW - T 12
SW - F FW - T 7 SW - T FW - T 11
SW - F + SW - T FW - F + FW - T 21
SW - F SW - T 12
FW - F FW - T 7
.. ' ....
SW - F = fully adapted salt water (17 ppt)
SW - T = salt water (17 ppt) samples i hr.
FW - F = fully adapted freshwater samples '
FW - T = freshwater samples i hr. to 24 hr.
* Significant at the .001 level ** Significant at the .01 level
T
*6.037 * s.102 *6.292
**3.108 0.002 0.244
*4.450 0.360 0.927
samples to J6 hr.
29
.. Table 4. The mean specific activity + s. E. of
++ ··-Mg - dependent ATPase in gill homogenates (2%) of !· heteroclitus.
Salinity
FW (fully adapted) FW Ci hr. to 24 hr.) FW (fully adapted +
i hr. to 24 hr.)
SW .(..fully adapted) SW (j hr. to 36 hr.)
SW (fully adapted + i hr~ to 36 hr.)
FW = freshwater
Number of samples
s 4
9 4 9
1J
SW = salt water (17 ppt)
umoles Pi/mg/min X ± S. E.
.1193 + .0203
.12,54 + .0~55 -
.1220 + .0125 -
.0908 + .0159 -
.1429 + .0086 -
.1269 + .0101 -
JO
Table 5. Results of T - test for significance between ++ means of Mg - dependent ATPase in gill
. . homogenates (2%) of F. heteroclitus.
x df T
SW .. F FW - F 8 0.056 SW - T FW -·F 12 1.256 SW - F + SW - T FW - F 17 0.371 SW - F FW - F + FW - T 11 1.439 SW - F FW - T ·6 1.553 SW - T FW - T 11 1.065
SW - F + SW - T FW - F + FW - T 21 0.363 SW - F SW - T . 11 *J,1J6
SW - F = fully adapted salt water (17 ppt) samples SW - T = salt water (17 ppt) samples i hr. to 36 hr. FW - F = fully adapted freshwater samples FW - T = freshwater samples i hr. to 24 hr.
* Significant at the .01 level
31
Table 6, Ouabain inhibition of (Na++K+) - dependent ATPase in gill preparations of F. heteroclitus.
Source of micro so mes
'·'"·-"'···
FW
SW
FW = freshwater
100 30 130 0
100 30
100 JO 130 0 100 30
SW = salt water (17 ppt)
Ouabain ATPase activity (mM) umoles Pi/mg/min
0 .788 1 .178 1 .179
0 .472 1 .176 1 .176
32
Table 7. Specific activities of (Na++K+) - dependent ·"ATPase and Mg++ - dependent ATPase in microsomal gill fractions of r. heteroclitus.
Salinity
FW
SW - 17 ppt FW
SW - 24 ppt FW SW - JO ppt FW
(Na++K+) - ATPase umoles Pi/mg/min
.610
.296 • 550 .200 .540 .985 .565
SW - 17 ppt, ! hr. ,770
. - .....
FW = freshwater SW = salt water
Mg++ - ATPase umoles Pi/mg/min
.178
.176
.177
.191
.144
.193
.233
.194
JJ
Table 8. The mean specific activity + s. E. of ( + +} - ++
Sample
FW
FW
Na +K - dependent ATPase and Mg -dependent ATPase in microsomal gill preparations of F. heteroclitus.
Number of samples
4 4
ATPase activity umoles Pi/mg/min
(Na++K+}-dependent Mg++-dependent
X + S. E.
• 5662 ± -. 01.54 .1830 + .0184
FW = freshwater
34
Table 9. Results of T - test for significance between means of Mg++ - dependent ATPase and (Na•+K+) - dependent ATPase in microsomal gill preparations or !· heteroclitus.
ATPase
Mg++-dependent
dr
SW(17 ppt, 6 1? ppt i hr ••
24'ppt, and
JO ppt)
(Na•+K•)-dependent FW SW(17 ppt, 6 "'l?_ppt i hr ••
24 ppt, and
JO ppt)
* Significant at the .001 level
T
.2907
* 15.9242
35
Table 10. uumoles JH - ouabain bound per mg protein in microsomal gill preparations of F. heteroclitus.
>-. ......... ... ·- m-> ·- E ... .09 u <( .........
·-u 0.. ·- \n '4- Q) u - 06 Q) 0 Q. E
U') :::>
. 03
_I_ ·T
SW (5)
T .. -i
SWT (9)
;-T
FW (5)
-r
FWT (4)
(Na++ K+)-dependent AT Pase
... .. .. . .. .. .. . . ..
:::~~::: ..... e e e ' I ..... .....
·.·.·.·.·.·~ ...... I ••••• ..... :::::~ ..... ...... . . . . . . ...... ...... ...... ..... I I I I I I
I I I I I ........ .........
SWr. (4)
.T
SWT (9)
FW (5)
FWT (4)
M ++ g - dependent A TPase
w
°'
Figure 2. uumoles of. )H - ouabain bound per mg protein - + + and specific activity of the (Na +K ) dependent ATPase in microsomal gill preparations of F. heteroclitus.
SW = salt water adapted fish FW = freshwater adapted fish ( ) = salinity in__ppt or number of
<( ...... ...... u ...... ....... ·-. ..... . . . . . . . .... ...... .14 ·-...... u ...... ..... C1> ...... ..... c. ...... ..... ...... en ····· ······ ..... ...... . .....
·.·.·.···!i:
40
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' ' ' ~
46
VITA
Mary Elizabeth Gilman was born in Richmond,
Virginia, December, 26, 1949. She attended Hanover
County Public Schools and was graduated from Patrick
Henry High School in Ashland, Virginia in June, 1967.
She received a Bachelor of Science degree from
Westhampton College of the University of Richmond in
June, 1971 with a major in biology and sociology.
While at Westhampton College she was a member of Phi
Beta Kappa, Beta Beta Beta, Eta Sigma Phi, and Kappa
Delta Pi honorary societies. After graduation, she
entered the University of Richmond Graduate School in
September, 1971. While at the University of Richmond
Graduate School she has become a member of Gamma
Sigma Epsilon, has been the recipient of a Williams
Fellowship, and has served as vice president of Beta
Beta Beta. She has also served as a General Biology,
Molecular Biology, Cell Physiology, Genetic8.>and
Organic Chemistry laboratory assistant during her
graduate study program. She completed the requirements
for the Master of Science Degree from the University
of Richmond in May, 1973. Future plans are uncertain.