Phenotypic developmental plasticity induced by preincubation egg storage in chicken embryos (Gallus gallus domesticus).

The developing chicken blastoderm can be temporarily maintained in dormancy below physiological zero temperature. However, prolonged preincubation egg storage impairs normal morphological and physiological development of embryos in a potential example of fetal programming (in this case, "embryonic programming"). We investigated how preincubation egg storage conditions (temperature, duration, hypoxia, and hypercapnia) affects viability, body mass, and physiological variables and functions in day 15 chicken embryos. Embryo viability was impaired in eggs stored for 2 and 3 weeks, with the effects greater at 22°C compared to 15°C. However, embryo size was reduced in eggs stored at 15°C compared with 22°C. Phenotypic change resulting from embryonic programming was evident in the fact that preincubation storage at 15°C diminished hematocrit (Hct), red blood cell concentration ([RBC]), and hemoglobin concentration ([Hb]). Storage duration at 15°C more severely affected the time course (2, 6, and 24 h) responses of Hct, [RBC], and [Hb] to progressive hypoxia and hypercapnia induced by submersion compared with storage duration at 22°C. The time-specific regulation of acid-base balance was changed progressively with storage duration at both 22 and 15°C preincubation storages. Consequently, preincubation egg storage at 22°C resulted in poor viability compared with eggs stored at 15°C, but size and physiological functions of embryos in eggs stored for 1-2 weeks were worse in eggs stored in the cooler than stored under room conditions. Avian eggs thus prove to be useful for examining developmental consequences to physiology of altered preincubation thermal environment in very early stages of development (embryonic programming).

Reduced heart rate and cardiac output differentially affect angiogenesis, growth, and development in early chicken embryos (Gallus domesticus).

An increase in both vascular circumferential tension and shear stress in the developing vasculature of the chicken embryo has been hypothesized to stimulate angiogenesis in the developing peripheral circulation chorioallantoic membrane (CAM). To test this hypothesis, angiogenesis in the CAM, development, and growth were measured in the early chicken embryo, following acute and chronic topical application of the purely bradycardic drug ZD7288. At hour 56, ZD7288 reduced heart rate (f(H)) by ~30% but had no significant effect on stroke volume (~0.19 ± 0.2 µL), collectively resulting in a significant fall in cardiac output (CO) from ~27 ± 3 to 18 ± 2 µL min(-1). Mean f(H) at 72 h of development was similarly significantly lowered by acute ZD7288 treatment (250 µM) to 128 ± 0.3 beats min(-1), compared with 174.5 ± 0.3 and 174.7 ± 0.8 beats min(-1) in control and Pannett-Compton (P-C) saline-treated embryos, respectively. Chronic dosing with ZD7288-and the attendant decreases in f(H) and CO-did not change eye diameter or cervical flexion (key indicators of development rate) at 120 h but significantly reduced overall growth (wet and dry body mass decreased by 20%). CAM vessel density index (reflecting angiogenesis) measured 200-400 µm from the umbilical stalk was not altered, but ZD7288 reduced vessel numbers-and therefore vessel density-by 13%-16% more distally (500-600 µm from umbilical stalk) in the CAM. In the ZD7288-treated embryos, a decrease in vessel length was found within the second branch order (~300-400 µm from the umbilical stock), while a decrease in vessel diameter was found closer to the umbilical stock, beginning in the first branch order (~200-300 µm). Paradoxically, chronic application of P-C saline also reduced peripheral CAM vessel density index at 500 and 600 µm by 13% and 7%, respectively, likely from washout of local angiogenic factors. In summary, decreased f(H) with reduced CO did not slow development rate but reduced embryonic growth rate and angiogenesis in the CAM periphery. This study demonstrates for the first time that different processes in the ontogeny of the early vertebrate embryo (i.e., hypertrophic growth vs. development) have differential sensitivities to altered convective blood flow.