title: Waste nitrogen metabolism and excretion in zebrafish embryos: effects of light, ammonia, and nicotinamide.
abstract: Bony fish primarily excrete ammonia as adults however the persistence of urea cycle genes may reflect a beneficial role for urea production during embryonic stages in protecting the embryo from toxic effects of ammonia produced from a highly nitrogenous yolk. This study aimed to examine the dynamic scope for changes in rates of urea synthesis and excretion in one such species (zebrafish, Danio rerio) by manipulating the intrinsic developmental rate (by alteration of light:dark cycles), as well as by direct chemical manipulation via ammonia injection (to potentially activate urea production) and nicotinamide exposure (to potentially inhibit urea production). Continuous dark exposure delayed development in embryos as evidenced by delayed appearance of hallmark anatomical features (heartbeat, eye pigmentation, body pigmentation, lateral line, fin buds) at 30 and 48 hr post-fertilization, as well by a lower hatching rate compared to embryos reared in continuous light. Both ammonia and urea excretion were similarly effected and were generally higher in embryos continuously exposed to light. Ammonia injection resulted in significant increases (up to fourfold) of urea N excretion and no changes to ammonia excretion rates along with modest increases in yolk ammonia content during 2-6 hr post-injection. Nicotinamide (an inhibitor of urea synthesis in mammals) reduced the ammonia-induced increase in urea excretion and led to retention of ammonia in the yolk and body of the embryo. Our results indicate that there is a relatively rapid and large scope for increases in urea production/excretion rates in developing embryos. Potential mechanisms for these increases are discussed.
authors.affiliation: Department of Biology, University of Ottawa, Ottawa, ON, Canada. carolbucking@gmail.com
authors.forename: Carol
authors.initials: C
authors.lastname: Bucking
authors.forename: Christophe M R
authors.initials: CM
authors.lastname: Lemoine
authors.forename: Patrick J
authors.initials: PJ
authors.lastname: Walsh
mesh_terms: D000641:Ammonia / Q000008:administration & dosage; D000818:Animals; D004625:Embryo, Nonmammalian / Q000187:drug effects* / Q000378:metabolism; D008027:Light; D018482:Muscle, Skeletal / Q000187:drug effects / Q000378:metabolism; D009536:Niacinamide / Q000494:pharmacology; D009584:Nitrogen / Q000737:chemistry*; D014508:Urea / Q000737:chemistry / Q000378:metabolism*; D015027:Zebrafish / Q000254:growth & development*
chemical_list: D009536:Niacinamide; D000641:Ammonia; D014508:Urea; D009584:Nitrogen