Combined use of immunocytochemistry and in situ hybridization to study β thyroid-stimulating hormone gene expression in pituitaries of hypothyroid rats

1990 ◽  
Vol 4 (5) ◽  
pp. 385-396 ◽  
Author(s):  
Jennifer H. Steel ◽  
Domhnall J. O'Halloran ◽  
Philip M. Jonesa ◽  
Susan Van Noorden ◽  
William W. Chin ◽  
...  
2006 ◽  
Vol 1118 (1) ◽  
pp. 232-238 ◽  
Author(s):  
Corinna Haupt ◽  
Else A. Tolner ◽  
Uwe Heinemann ◽  
Otto W. Witte ◽  
Christiane Frahm

1997 ◽  
Vol 272 (2) ◽  
pp. E212-E217 ◽  
Author(s):  
S. Suwanwalaikorn ◽  
M. Van Auken ◽  
M. I. Kang ◽  
S. Alex ◽  
L. E. Braverman ◽  
...  

We have previously reported that thyroid-stimulating hormone (TSH)-suppressive doses of L-thyroxine (L-T4) decrease femoral, but not vertebral, bone mineral density (BMD) in rats. L-T4-induced decreases in BMD were associated with increased expression of genes, reflecting osteoblast activity in mRNA extracted from whole femurs but not from vertebrae. To document that this skeletal selectivity reflected altered osteoblast activity, we studied gene expression by in situ hybridization in 8-wk-old rats treated with L-T4 (20 microg x 100 g body wt(-1) x day(-1)) for 4 wk. TSH-suppressive doses of L-T4 were associated with decreased femoral (0.299 +/- 0.005 vs. 0.273 +/- 0.005 g/cm2, P < 0.01), but not vertebral (0.222 +/- 0.004 vs. 0.218 +/- 0.003 g/cm2), BMD. In situ hybridization documented that L-T4 administration for 4 wk increased expression of osteocalcin and alkaline phosphatase mRNA in femoral, but not vertebral, osteoblasts. This study demonstrates a differential gene expression response of vertebral and femoral osteoblasts to L-T4. This altered degree of gene expression markers of osteoblast activity documented by in situ hybridization may in part explain the apparent clinical differences in the effect of L-T4 on femoral and vertebral BMD.


2021 ◽  
Vol 4 (1) ◽  
pp. 20
Author(s):  
Mujeeb Shittu ◽  
Tessa Steenwinkel ◽  
William Dion ◽  
Nathan Ostlund ◽  
Komal Raja ◽  
...  

RNA in situ hybridization (ISH) is used to visualize spatio-temporal gene expression patterns with broad applications in biology and biomedicine. Here we provide a protocol for mRNA ISH in developing pupal wings and abdomens for model and non-model Drosophila species. We describe best practices in pupal staging, tissue preparation, probe design and synthesis, imaging of gene expression patterns, and image-editing techniques. This protocol has been successfully used to investigate the roles of genes underlying the evolution of novel color patterns in non-model Drosophila species.


Genetics ◽  
2021 ◽  
Vol 217 (1) ◽  
Author(s):  
Amir Fallahshahroudi ◽  
Martin Johnsson ◽  
Enrico Sorato ◽  
S J Kumari A Ubhayasekera ◽  
Jonas Bergquist ◽  
...  

Abstract Domestic chickens are less fearful, have a faster sexual development, grow bigger, and lay more eggs than their primary ancestor, the red junglefowl. Several candidate genetic variants selected during domestication have been identified, but only a few studies have directly linked them with distinct phenotypic traits. Notably, a variant of the thyroid stimulating hormone receptor (TSHR) gene has been under strong positive selection over the past millennium, but it’s function and mechanisms of action are still largely unresolved. We therefore assessed the abundance of the domestic TSHR variant and possible genomic selection signatures in an extensive data set comprising multiple commercial and village chicken populations as well as wild-living extant members of the genus Gallus. Furthermore, by mean of extensive backcrossing we introgressed the wild-type TSHR variant from red junglefowl into domestic White Leghorn chickens and investigated gene expression, hormone levels, cold adaptation, and behavior in chickens possessing either the wild-type or domestic TSHR variant. While the domestic TSHR was the most common variant in all studied domestic populations and in one of two red junglefowl population, it was not detected in the other Gallus species. Functionally, the individuals with the domestic TSHR variant had a lower expression of the TSHR in the hypothalamus and marginally higher in the thyroid gland than wild-type TSHR individuals. Expression of TSHB and DIO2, two regulators of sexual maturity and reproduction in birds, was higher in the pituitary gland of the domestic-variant chickens. Furthermore, the domestic variant was associated with higher activity in the open field test. Our findings confirm that the spread of the domestic TSHR variant is limited to domesticated chickens, and to a lesser extent, their wild counterpart, the red junglefowl. Furthermore, we showed that effects of genetic variability in TSHR mirror key differences in gene expression and behavior previously described between the red junglefowl and domestic chicken.


2004 ◽  
Vol 229 (3) ◽  
pp. 651-657 ◽  
Author(s):  
Nathaniel Denkers ◽  
Pilar García-Villalba ◽  
Christopher K. Rodesch ◽  
Kandice R. Nielson ◽  
Teri Jo Mauch

1991 ◽  
Vol 88 (18) ◽  
pp. 8154-8158 ◽  
Author(s):  
A. M. Henney ◽  
P. R. Wakeley ◽  
M. J. Davies ◽  
K. Foster ◽  
R. Hembry ◽  
...  

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