Comparing techniques for measuring corticosterone in tadpoles

نویسندگان

  • Pablo BURRACO
  • Rosa ARRIBAS
  • Saurabh S. KULKARNI
  • Daniel R. BUCHHOLZ
  • Ivan GOMEZ-MESTRE
چکیده

Glucocorticoids play a key role in mediating stress responses in vertebrates. Corticosterone (CORT) is the main glucocorticoid produced in amphibians, birds, and reptiles, and regulates several metabolic functions. The most common methods for quantifying CORT are competitive binding immunoassays: radioimmunoassay (RIA) and enzyme immunoassay (EIA). RIA has been broadly used since the 1980’s but it requires radioactivity. Commercial EIA kits permit quantifying hormone levels without radioactivity although the requirement for a larger sample volume may be a strong limitation for measurements involving larval amphibians. Here we used Xenopus laevis tadpoles to compare the performance of three commonly used procedures for determination of CORT: RIA on a chloroform extract of whole-body homogenate, EIA on plasma, and EIA on supernatant of whole-body homogenate. We treated tadpoles with exogenous CORT at 0, 25, 50, and 100 nM. RIA could distinguish between 0 and 25 nM, and EIA on plasma between 0 and 50 nM, whereas whole-body homogenate EIA only detected significant differences between 0 and 100 nM. Each procedure presents advantages and disadvantages regarding sensitivity, the use of radioactivity, sample size, handling time, and economic cost. RIA is preferred when studying small-bodied animals from which blood samples cannot be obtained. When CORT level differences are intermediate and blood sampling is possible, EIA on plasma is a good non-radioactive alternative. EIA on whole-body homogenates may be useful to assess qualitative changes in CORT levels when considerable differences are expected. Finally, we discuss our findings in the context of previous studies on CORT in amphibians [Current Zoology 61 (5): 835–845, 2015].

برای دانلود رایگان متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Survival, the hormonal stress response and UV-B avoidance in Cascades Frog tadpoles ( Rana cascadae ) exposed to UV-B radiation

1. Despite the increasing occurrence of global environmental changes, including increases in ultraviolet-B radiation (UV-B; 280–320 nm), little is known about how factors such as UV-B affect animals physiologically. Amphibians provide a good model for examining physiological effects of UV-B exposure because studies documenting both lethal and sub-lethal effects have been completed on many speci...

متن کامل

ARPN Journal of Science and Technology:: Corticosterone as Inhibitor of Bufo Melanostictus Tadpole Growth

This investigation reports the effect of corticosterone on the growth of Bufo melanostictus tadpoles. Test tadpoles of experiment I (T-I), maintained in 1μg/ml solution of corticosterone, were 2.39mm shorter than controls by day 11. Test tadpoles of experiment II (T-II), maintained in 5μg/ml solution were 4.32mm shorter than their counterparts. Growth inhibition was dose related and became sign...

متن کامل

Corticosterone and Growth in Pacific Treefrog (Hyla regilla) Tadpoles

In all vertebrates, responses to environmental perturbation are regulated to at least some degree by glucocorticoid hormones (GCs). In amphibians, GCs, along with several other hormones, also play an important role in larval growth and development preceding metamorphosis. Given these two well-established functions, GCs might be expected to have a role in the well-documented effects that environ...

متن کامل

The influence of visual and tactile stimulation on growth and metamorphosis in anuran larvae

1. Sensory modalities that allow tadpoles to assess their environment, and subsequently mediate their development, are not well understood. 2. By putting clay model tadpoles into the tanks with live tadpoles we have enhanced tactile and visual stimuli for tadpoles of three species ( Rana sylvatica , Bufo americanus and Xenopus laevis ) in a controlled fashion. The goal was to determine whether ...

متن کامل

Roles of stress hormones in food intake regulation in anuran amphibians throughout the life cycle.

Towards understanding the ontogeny of energy balance regulation in vertebrates we analyzed the responses of corticotropin-releasing factor (CRF) and corticosterone to food deprivation in the Western spadefoot toad (Spea hammondii) at three developmental stages: premetamorphic tadpole, prometamorphic tadpole, and juvenile. Corticosterone responses to 5 days of food deprivation varied among devel...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

عنوان ژورنال:

دوره   شماره 

صفحات  -

تاریخ انتشار 2014