Effect of long-term oral administration of extra thyroxine on oviductal expression of carbonic anhydrase and avidin-related protein-2 genes in broiler breeder hens

Document Type : Short Communication


1 Department of Animal Science, Faculty of Animal Science and Fishery, Sari Agricultural Sciences and Natural Resources University, Sari, Iran.

2 Department of Animal Science, Faculty of Agriculture, Shiraz University, Shiraz, Iran.


Avian sperm are stored in the sperm storage tubules (SSTs) of the hen oviduct for a prolonged period. The impact of avidin-related protein-2 (AVRP2) and carbonic anhydrase II (CA II) in sperm viability in the SSTs has been suggested. The aim of the present study was to investigate the effect of oral administration of a high dose of thyroxine on the oviductal expression of AVRP2 and CA II genes in broiler breeder hens. The birds (n=70), housed in separate cages, were randomly allotted to two treatment groups to either zero (CON) or 0.30 mg thyroxine per day (T4 group) for 12 weeks. Feed and water were supplied according to the Cobb 500 standards (metabolizable energy: 2700 kcal/kg and crude protein: 14%). Blood samples were prepared seven times for determination of plasma triiodothyronine (T3) and T4 concentration. At the end of the treatment period, 20 hens were randomly selected and killed to determine the expression of AVRP2 and CA II in the SSTs using the real-time PCR procedures. Expressions of AVRP2 and CA II genes were influenced by T4 treatment where an increased expression of CA II was recorded for T4-exposed hens (P<0.05). However, expression of AVRP2 was not significantly different between the treatment groups (P>0.05).


Main Subjects

  • Akhlaghi, A., Zamiri, M.J., Ahangari, Y.J., Atashi, H., Pirsaraei, Z.A., Deldar, H., Hashemi, S.R. 2013. Oral exposure of broiler breeder hens to extra thyroxine modulates early adaptive immune responses in progeny chicks. Poultry Science 92, 1040-1049.
  • Akhlaghi, A., Zamiri, M.J., Shahneh, A.Z., Ahangari, Y.J., Javaremi, A.N., Mianji, G.R., Atashi, H. 2012. Maternal hyperthyroidism is associated with a decreased incidence of cold-induced ascites in broiler chickens. Poultry Science 91, 1165-1172.‏
  • Bakst, M.R., Donoghue, A.M., Yoho, D.E., Moyle, J.R., Whipple, S.M., Camp, M.J., Bramwell, R.K. 2010. Comparisons of sperm storage tubule distribution and number in 4 strains of mature broiler breeders and in turkey hens before and after the onset of photostimulation. Poultry Science 89, 986-992.
  • Bialy, G., Pincus, G. 1962. Effects of estrogen and progestin on the uterine carbonic anhydrase of immature rats. Endocrinology 70, 781-785.
  • ‏Board, R.G., Fuller, R. 1974. Non‐specific antimicrobial defences of the avian egg embryo and neonate. Biological Reviews 49, 15-49.‏
  • Chan, L., Means, A.R., O'Malley, B.W. 1973. Rates of induction of specific translatable messenger RNAs for ovalbumin and avidin by steroid hormones. Proceedings of the National Academy of Sciences 70, 1870-1874.‏
  • Christensen, V.L., Bagley, L.G. 1989. Efficacy of fertilization in artificially-inseminated turkey hens. Poultry Science 68, 724-729.‏
  • Comstock, J.P., Rosenfeld, G.C., O'Malley, B.W., Means, A.R. 1972. Estrogen-induced changes in translation, and specific messenger RNA levels during oviduct differentiation. Proceedings of the National Academy of Sciences 69, 2377-2380.
  • Daryabari, H., Akhlaghi, A., Zamiri, M.J., Pirsaraei, Z.A., Mianji, G.R., Deldar, H., Eghbalian, A.N. 2015. Oral administration of supplementary biotin differentially influences the fertility rate and oviductal expression of avidin and avidin-related protein-2 in low-and high-fertility broiler line hens. Poultry science 94, 289-295.
  • Das, S.C., Isobe, N., Nishibori, M., Yoshimura, Y. 2006a. Expression of transforming growth factor-β isoforms and their receptors in utero-vaginal junction of hen oviduct in presence or absence of resident sperm with reference to sperm storage. Reproduction 132, 781-790.‏
  • Das, S.C., Nagasaka, N., Yoshimura, Y. 2006b. Changes in the expression of estrogen receptor mRNA in the utero-vaginal junction containing sperm storage tubules in laying hens after repeated artificial insemination. Theriogenology 65, 893-900.
  • Dobyan, D.C., Bulger, R.E. 1982. Renal carbonic anhydrase. American Journal of Physiology-Renal Physiology 243, 311-324.
  • Elo, H.A. 1980. Occurrence of avidin-like biotin-binding capacity in various vertebrate tissues and its induction by tissue injury. Comparative Biochemistry and Physiology Part B: Comparative Biochemistry 67, 221-224.‏
  • Foye-Jackson, O.T., Long, J.A., Bakst, M.R., Blomberg, L.A., Akuffo, V.G., Silva, M.V.B., McMurtry, J.P. 2011. Oviductal expression of avidin, avidin-related protein-2, and progesterone receptor in turkey hens in relation to sperm storage: Effects of oviduct tissue type, sperm presence, and turkey line. Poultry Science 90, 1539-1547.
  • Gope, M.L., Keinānen, R.A., Kristn, P.A., Conneely, O.M., Beattie, W.G., Zarucki-Schulz, T., Kulomaa, M.S. 1987. Molecular cloning of the chicken avidin cDNA. Nucleic Acids Research 15, 3595-3606.‏
  • Hemmings, N., Birkhead, T.R., Brillard, J.P., Froment, P., Brière, S. 2015. Timing associated with oviductal sperm storage and release after artificial insemination in domestic hens. Theriogenology 83, 1174-1178.
  • Hodgen, G.D., Falk, R.J. 1971. Estrogen and progesterone regulation of carbonic anhydrase isoenzymes in guinea pig and rabbit uterus. Endocrinology 89, 859-864.‏
  • Holm, L., Ridderstraale, Y. 1998. Localization of carbonic anhydrase in the sperm-storing regions of the turkey and quail oviduct. The Histochemical Journal 30, 481-488.‏
  • Holm, L., Ridderstraale, Y., Knutsson, P.G. 1996. Localisation of carbonic anhydrase in the sperm storing regions of the domestic hen oviduct. Cells Tissues Organs 156, 253-260.
  • ‏‏Holm, L., Ruziwa, S.D., Dantzer, V., Ridderstraale, Y. 2000. Carbonic anhydrase in the utero-vaginal junction of immature and mature ostriches. British Poultry Science 41, 244-249.‏
  • Kunnas, T.A., Joensuu, T.K., Viitala, K.K., Sopanen, P., Tuohimaa, P., Kulomaa, M.S. 1992. Induction of avidin messenger ribonucleic acid in the chick oviduct by progesterone and other steroids. Endocrinology 130, 3421-3426.‏
  • Kunnas, T.A., Wallén, M.J., Kulomaa, M.S. 1993. Induction of chicken avidin and related mRNAs after bacterial infection. Biochimica et Biophysica Acta (BBA)-Gene Structure and Expression 1216, 441-445.
  • Kuramitz, H., Matsuda, M., Sugawara, K., Tanaka, S. 2003. Electrochemical evaluation of the interaction between avidin and biotin at biotinylated polypyrrole electrode using a redox marker. Electroanalysis 15, 225-229.‏
  • Kurył, J. 1981. Carbonic anhydrase activity in the membrane and contents of the ovarian follicles and oviduct mucosa of hens. Acta Physiologica Polonica 33, 279-285.‏
  • Laitinen, O.H., Hytönen, V.P., Ahlroth, M.K., Pentikäinen, O.T., Gallagher, C., Nordlund, H.R., Johnson, M.S. 2002. Chicken avidin-related proteins show altered biotin-binding and physico-chemical properties as compared with avidin. Biochemical Journal 363, 609-617.‏
  • Li, Z., Yang, L., Wang, J., Shi, W., Pawar, R.A., Liu, Y., Xia, F. 2010. β-actin is a useful internal control for tissue-specific gene expression studies using quantitative real-time PCR in the half-smooth tongue sole cynoglossus semilaevis challenged with LPS or Vibrio anguillarumFish & Shellfish Immunology 29, 89-93.‏
  • Livak, K.J., Schmittgen, T.D. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. Methods 25, 402-408.‏
  • Maren, T.H. 1967. Carbonic anhydrase: chemistry, physiology, and inhibition. Physiological Reviews 47, 595-781.‏
  • McNabb, F.A. 2007. The hypothalamic-pituitary-thyroid (HPT) axis in birds and its role in bird development and reproduction. Critical Reviews in Toxicology 37, 163-193.‏
  • Means, A. R., Comstock, J. P., Rosenfeld, G. C., O'Malley, B. W. 1972. Ovalbumin messenger RNA of chick     oviduct: partial characterization, estrogen dependence, and translation in vitro. Proceedings of the National Academy of Sciences 69, 1146-1150.‏
  • Miyake, T., Pincus, G. 1959. Hormonal influences on the carbonic anhydrase concentration in the accessory reproductive tracts of the rat. Endocrinology 65, 64-72.
  • Nys, Y., Parkes, C.O., Thomasset, M. 1986. Effects of suppression and resumption of shell formation and parathyroid hormone on uterine calcium-binding protein, carbonic anhydrase activity, and intestinal calcium absorption in hens. General and Comparative Endocrinology 64, 293-299.‏
  • Pierson, E.E.M., McDaniel, G.R., Krista, L.M. 1988. Relationship between fertility duration and in vivo sperm storage in broiler breeder hens. British Poultry Science 29, 199-203.‏
  • SAS. 2003. User’s Guide: Statistics, Version 9.1 Edition. Inst., Inc., Cary, NC.
  • Sechman, A. 2013. The role of thyroid hormones in regulation of chicken ovarian steroidogenesis. General and Comparative Endocrinology 190, 68-75.‏
  • Sechman, A., Paczoska-Eliasiewicz, H., Rzasa, J., Hrabia, A. 2000. Simultaneous determination of plasma ovarian and thyroid hormones during sexual maturation of the hen (Gallus domesticus). Folia-Biologica-Krakow 48, 7-12.‏
  • Sechman, A., Pawlowska, K., Rzasa, J. 2009. Influence of triiodothyronine (T3) on secretion of steroids and thyroid hormone receptor expression in chicken ovarian follicles. Domestic Animal Endocrinology 37, 61-73.‏
  • Tuohimaa, P., Joensuu, T., Isola, J., Keinanen, R., Kunnas, T., Niemela, A., Kulomaa, M. 2003. Development of progestin-specific response in the chicken oviduct. International Journal of Developmental Biology 33, 125-134.
  • Yoshimura, Y., Koike, K., Okamoto, T. 2000. Immunolocalization of progesterone and estrogen receptors in the sperm storage tubules of laying and diethylstilbestrol-injected immature hens. Poultry Science 79, 94-98.