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| Status |
Public on Jul 31, 2013 |
| Title |
Effects of 5α-dihydrotestosterone on male mummichog (Fundulus heteroclitus) testes steroid production and gene expression patterns |
| Organism |
Fundulus heteroclitus |
| Experiment type |
Expression profiling by array
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| Summary |
To better assess the impact of contaminants on fish reproduction, it is increasingly important to collect data at multiple levels of biological organization. However, a challenge in ecotoxicology is that three of the most commonly measured reproductive endpoints; gonad histology, ex vivo steroid production, and gene expression, can be difficult to obtain in small-bodied fish (e.g. zebrafish, Japanese medaka, fathead minnow) due to limited tissue availability. Using 5α-dihydrotestosterone (DHT) as a test compound, the objectives of the current study were to assess the feasibility of measuring gene expression in tissues also used for ex vivo steroid production studies. A portion of the testes was also used for qualitative histological examination. Male mummichog were exposed to 0, 5, and 50 µg/L DHT for 21 days. Following the exposure, testes were collected and separated into three pieces for histology, ex vivo incubation to measure steroidogenic capacity [Testosterone(T), 17β-estradiol (E2), 11-ketotestosterone (11KT)] and gene expression analyses. Testis RNA at time of dissection and testis RNA at end of ex vivo incubation (18 h) was collected and assessed for RNA integrity. The expression of genes coding for estrogen receptor βa (esrba) and estrogen receptor βb (esrbb) as well as the key steroidogenic genes, steroidogenic acute regulatory protein (star), P450 side chain cleavage (p450scc) and 11β-hydroxysteroid dehydrogenase (hsd11b3) were assessed in testis before and after ex vivo incubation. There were no significant differences in RNA quality between the two time points, however the expression of esrba, esrbb and p450scc was significantly lower post-incubation, although star, hsd11b3, and all four reference gene expression levels remained unchanged over time. DHT did not affect the expression of any of the measured genes at either concentration. Histological examination showed that all fish contained mature testes dominated by spermatozoa and spermatids. DHT did not affect T production but significantly decreased 11KT production in the high treatment group. This suggests a role for DHT in androgen regulation, and has
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| Overall design |
Microarray analysis was performed with four biological replicates (4 control and 4 treatment). Male mummichog were exposed to 0, 5, and 50 µg/L DHT for 21 days. Individuals from the high concentration, 50 ug/L DHT, were used for the microarray analysis.
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| Contributor(s) |
Martyniuk C, Ings J, Doyle M, Feswick A, Bosker T, Munkittrick K |
| Citation missing |
Has this study been published? Please login to update or notify GEO. |
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| Submission date |
Jul 08, 2013 |
| Last update date |
Aug 02, 2013 |
| Contact name |
Christopher Joseph Martyniuk |
| E-mail(s) |
CMARTYN@UFL.EDU
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| Organization name |
UF
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| Department |
Physiological Sciences
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| Street address |
CEHT Mowry Road
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| City |
Gainesville |
| State/province |
FL |
| ZIP/Postal code |
32611 |
| Country |
USA |
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| Platforms (1) |
| GPL17413 |
Agilent-027999 Custom Fundulus heteroclitus array |
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| Samples (8)
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| GSM1181820 |
Testis, DHT50, Rep3 |
| GSM1181821 |
Testis, Control, Rep2 |
| GSM1181822 |
Testis, DHT50, Rep4 |
| GSM1181823 |
Testis, Control, Rep3 |
| GSM1181824 |
Testis, Control, Rep4 |
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| Relations |
| BioProject |
PRJNA210759 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE48582_RAW.tar |
8.6 Mb |
(http)(custom) |
TAR (of TXT) |
| Processed data included within Sample table |
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