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Ovarian cortical tissues of various ages of chicken were grafted underneath chorioallantoic membrane (CAM) of 6-d-old chicken embryo and the grafts were collected on d 0, 2, 4, 8, or 10. The tissue sections were prepared to examine the development of follicles in grafts and the proliferative ability of follicles was examined by immunohistochemistry analysis of proliferating cell nuclear antigen. The results indicated that the development of follicles in cultured chicken ovarian cortical pieces progressed smoothly and slowly similar to those in vivo, and the environment of chicken embryo was more suitable for the synchronous development of oocyte and follicular cells in the larger follicles. Meanwhile, the synchronous development could be more easily achieved in the chicken ovarian tissues of 12- to 15-wk chicken. Neither oocyte nor granulosa cells but the thecal cells of follicles developed in grafts of 13- to 15-d chicken within 10 d of culturing in ovo, and the CAM can ease follicular atresia in chicken ovarian grafts of 12- to 15-wk chicken. The proliferative performance of follicles in the grafts was not influenced by the environment of chicken embryo. These results indicated that the chicken embryo has the ability to support slow development of primordial and primary follicles in grafted ovarian cortical tissues of chickens of different ages. The CAM system may also prove useful for the study of early follicle development with further information.

An experiment was carried out to determine the effect of dietary protein level on the specific activity of hepatic L-threonine dehydrogenase in young growing chicks. Six replicate pens of seven Leghorn chicks were fed semipurified diets containing 23, 27, or 32% CP with identical relative proportions of amino acids in each protein group. Body weights and feed consumption were measured for 3 d, and hepatic mitochondria were isolated for assay of threonine dehydrogenase (TDH) activity. Weight gains and feed efficiency increased at each level of protein supplementation, but feed consumption was not affected by protein level. The specific activity of threonine dehydrogenase in isolated liver mitochondria was significantly (P < 0.05) higher in the 32% CP group than in the 23% CP group, and the activity in the 27% CP group was intermediate. We conclude that moderate increases in dietary protein level result in elevated hepatic threonine dehydrogenase activity in growing chicks.

Amino acid imbalances contribute to higher requirements of amino acids than would occur if the dietary profile of amino acids perfectly matched the requirements. The mechanisms of imbalances have not been fully elucidated. Because threonine dehydrogenase (TDH) activity in liver mitochondria increases in chicks and rats subjected to threonine imbalance, the current study was carried out to determine whether the change in TDH activity occurs rapidly enough after the consumption of an imbalanced diet to be considered a possible primary metabolic response. In a series of experiments, Leghorn chicks were allowed free access to a semipurified basal diet marginally limited in threonine or the same diet containing a mixture of indispensable amino acids (IAA) lacking threonine to cause a threonine imbalance. In the first experiment, dietary supplements of 5.5 and 11.1% IAA were used to determine a level of supplement that would cause a robust response in the specific activity of TDH. Feed intake, body weight gains and efficiency of feed utilization were lower and specific activities of TDH were higher in chicks fed 11.1% IAA than in those fed 5.5% IAA. In subsequent experiments, hepatic TDH activities and plasma amino acid profiles of the control and experimental groups were determined at 1. 5, 3, 6, 12 and 24 h after the first offering of the diet containing 11.1% IAA. The specific activities of TDH in chicks fed the IAA supplement were 40-150% higher (P < 0.05) and plasma threonine concentrations were 42-53% lower (P < 0.05) than in chicks fed the basal diet at all times except 1.5 h. These results indicate that changes in the capacity for threonine degradation via TDH may occur in the liver within a few hours after the consumption of a threonine-imbalanced diet and suggest the possibility that altered TDH activity may contribute to the increased threonine requirement associated with threonine imbalance.

Experiments were conducted to determine the conditions for assay of hepatic phenylalanine hydroxylase (PAH) activity in the chicken and to determine the developmental pattern of PAH activity in liver 25,000 x g supernatant. PAH activity was detected in liver supernatant and (postnuclear) 25,000 x g particulate fraction. Optimum assay conditions differed for the two cell fractions, the most notable difference being a broad pH optimum of 7.7 to 9.2 for the supernatant and 4.7 and 5.6 for the particulate fraction. The PAH activity in the supernatant increased to a maximum as L-phenylalanine concentration in the assay medium increased from 0.02 to 0.5 mM and 1.0 mM. Activity increased in the particulate fraction as the Phe concentration increased to 0.5 mM. Substrate inhibition of PAH activity occurred at Phe concentrations of 3 to 5 mM in the supernatant but not in the particulate fraction. Concentrations of the cofactor, 6(R)-5,6,7,8-tetrahydrobiopterin, ranging from 0.09 to 0.75 mM, resulted in maximal PAH activity. The developmental pattern of PAH in supernatant was determined using a modified assay in which substrate and cofactor concentrations and pH were optimum. The PAH activity in liver supernatant was present at a low level in 11 d chick embryos and increased several fold between Days 15 and 17 to a maximum at Days 17 to 21. Activity declined at hatching to levels that were present in 11 to 15 d embryos and remained at this level in male chicks through 4 wk of age. Mature males had higher PAH activity than mature laying females.