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Prediction of the Fat Content of Pork Carcasses Based on Cross-sectional Region Analysis of Dual Energy X- Ray Absorptiometry Scans

pituitary glands removed. Individual cells were dispersed with dispase and cells were maintained at 37C (humidified atmosphere; 95% air: 5% CO2) in Dulbecco modified Eagle’s medium for 24 h. Cells were then treated with one of the following (2 wells /treatment; replicated in 3 experiments): 1. controls; 2. 10 nM T3; 3. 50 M DTPA; 4. 10 nM T3 plus 50 M DTPA; and 5. 10 nM T3, 50 M DTPA and 40 M zinc sulfate. Cells were incubated for 48 h prior to RNA extraction. Reverse transcription (RT) PCR, with specific primers for GH and thyroid stimulating hormone (TSH), was used to measure mRNA levels. Comparisons with control gene-specific primers RPL-32 and β-actin were used to calculate mRNA levels of expression for GH and TSH, respectively. To evaluate RT-PCR, preliminary experiments were performed with GH3 cells. Results with RT-PCR and the GH primer paralleled previous results using Northern analysis, showing that expression DTPA and T3 stimulated GH mRNA expression. TSH was measured to confirm the viablity of the anterior pituitary cells. As expected, T3 inhibited TSH mRNA expression 50% compared to control cells. Expression of TSH mRNA was greater in DTPA treated cells compared with controls (p<.01), but was lowered by combination with T3 and by combination with T3 and zinc. These results confirm that the anterior pituitary cells are viable and responsive. In these same cells DTPA alone reduced GH mRNA levels by 25% (p=.03). However, in contrast to GH3 cells, DTPA did not stimulate GH mRNA levels in T3-treated pituitary cells (p=.18). In conclusion, while zinc chelation increases TSH mRNA, in the presence or absence of T3, it does not affect the GH mRNA in primary rat anterior pituitary cells in the same manner as GH3 cells.