We examined thyroidectomized chickens in terms of plasma lipid concentration and protein expression within the liver. Although the body weight of thyroidectomized chickens was remarkably low due to growth retardation, the livers were enlarged and fatty compared to those of sham-operated chickens. An increase in phospholipid, triglyceride, and total cholesterol levels within the blood plasma of thyroidectomized chickens was observed, clearly reflecting increased lipid synthesis within the liver. Overexpression of some proteins, for example, 29- and 45-kDa proteins, was observed in thyroidectomized chicken livers by means of electrophoresis. A peptide map was made for the protein that exhibited the greatest degree of overexpression. One of them demonstrated a molecular mass of 45 kDa and an isoelectric point (pI) between 7.5 and 8.0, depending on its form. Partial N-terminal amino acid sequences were determined from three random peptides of this protein. The amino acid sequence of this protein showed a high degree of homology with the betaine-homocysteine S-methyltransferase (BHMT, EC 2.1.1.5) of some mammalian species. We identified this protein as chicken BHMT because, in addition to its sequence homology with mammalian BHMT, there were similarities were also observed between this 45-kDa protein and mammalian BHMT with respect to molecular mass and isoelectric behavior. In the liver, 10 d after thyroidectomy, the synthesis of hepatic BHMT had already been enhanced, and the high expression was maintained at 50 d of age. Generally, BHMT catalyzes the transfer of a methyl group from betaine to L-homocysteine. In addition, it seems that this enzyme is also closely related to lipid metabolism in the liver; in this study expression of BHMT in the liver corresponded to plasma lipid levels. Moreover, hypothyroidism may be directly or indirectly related to overexpression of BHMT. Due to similarities between the BHMT of chickens and mammalian species, the chicken model might provide a useful means by which to study BHMT, its role in lipid metabolism, and methods of targeting the expression of BHMT. Another 29-kDa protein was unidentified in the homology search.
A technique for transbucal hypophysectomy of the fowl using a stereotaxic instrument is described. A total of 95 hens were hypophysectomized by our technique with an operating time of about 10 minutes per bird. About 8% of the birds died soon after the operation either as a result of injury to the brain or from hemorrhage or an overdose of Nembutal and 13% were found to be incompletely hypophysectomized. The results suggest that our new techinique for hypophysectomy of the fowl is usetul not only for inexperienced operators but also for technicians trained for orthodox hypophysectomy, since location of the pituitary is quite accessible and the instrument provides considerable accuracy in the subsequent drilling.
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