The Effect of Temperature on Angiogenesis in Chicken Embryos

Abstract

Angiogenesis plays an important role in the formation of new vessels in the embryonic period. The aim of this study is to determine possible differences in angiogenesis process in chicken embryos exposed to two different incubation temperatures of 37.50C and 39.00C using Chorioallantoic Membrane (CAM) model. The study included two groups, each with an experimental and control group, in which 16 fertilized chicken eggs were analyzed. The eggs were opened on the 7th day of incubation with the aid of forceps and surgical scissors without damaging the embryo. Head-stern, occipito-frontal length, vessel thickness, density and number of each embryo on CAM were measured by ImageJ. Statistical analysis of the data was performed by SPSS program. Depending on the temperature, there was a decrease, although not statistically significant, in the values of head-stern lengths of the embryos obtained from the eggs incubated at 39.00C. However, it was determined that occipito-frontal length values increased, the skull had a larger appearance and this increase was statistically significant (p<0.05). The mean density of embryos obtained from eggs incubated at 37.50C was 1.58±0.57, while the mean vessel density of embryos obtained from eggs incubated at 39.00C was 1.46±0.61. There was no statistically difference between mean vessel density values (p>0.05). A comparison of both groups showed a decrease in the number of vascular branches (35.44±10.97 at 37.50C, 24.94±6.97 at 39.00C) due to the increasing temperature. This decrease was found to be statistically significant (p<0.05). In general, it was observed that the increase in temperature caused a decrease in vessel density, thickness and number of branches in chicken embryos on CAM. As a result, it was determined that growth and development were negatively affected in embryos.

Keywords

• chorioallantoic membrane,Angiogenesis,chick,temperature,vessel

Öz

Anjiyogenez embriyonik dönemde yeni damarların oluşumunda önemli bir rol oynar. Bu çalışmanın amacı, Koryoallantoik Membran (KAM) modeli kullanılarak 37.50C ve 39.00C olacak şekilde iki farklı kuluçka sıcaklığına maruz bırakılan tavuk embriyolarında anjiyogenez sürecindeki olası farklılıkları belirlemektir. Çalışmada her birinde 16’şar adet döllü tavuk yumurtasının analiz edildiği deney ve kontrol grubu olmak üzere iki grup yer aldı. Yumurtalar inkübasyonun 7. gününde penset ve cerrahi makas yardımıyla embriyoya zarar vermeden açıldı. Fotoğrafları çekilen embriyoların tepe-kıç ve occipito-frontal uzunlukları ile KAM’daki damar kalınlıkları, damar yoğunluğu ve damar dal sayıları ImageJ programı kullanılarak ölçüldü. Verilerin istatistiksel analizi SPSS programı aracılığıyla yapıldı. Sıcaklığa bağlı olarak 39.00C’de inkübe edilen yumurtalardan elde edilen embriyoların tepe-kıç uzunluklarının değerlerinde istatistiksel olarak anlamlı olmasa da bir azalma mevcuttu. Bununla birlikte occipito-frontal uzunluk değerlerinin arttığı, kafatasının daha büyük bir görünüme sahip olduğu ve bu artışın istatistiksel olarak anlamlı olduğu tespit edildi (p<0.05). 37.50C’de inkübe edilen yumurtalardan elde edilen embriyoların damar yoğunluğu ortalaması 1.58±0.57 iken, 39.00C’de inkübe edilen yumurtalardan elde edilen embriyoların ortalama damar yoğunluğu 1.46±0.61 olarak tespit edildi. Ortalama damar yoğunluğu değerleri arasında istatistiksel olarak gruplar arasında bir fark gözlenmedi (p>0.05). Her iki grubun karşılaştırılmasında artan sıcaklığa bağlı olarak damar dal sayısında bir azalma (37.50C’de 35.44±10.97, 39.00C’de 24.94±6.97) olduğu görüldü. Bu azalmanın istatistiksel olarak anlamlı olduğu tespit edildi (p<0.05). Genel anlamda sıcaklık artışının tavuk embriyolarında KAM’da damar yoğunluğu, kalınlığı ve dal sayısında bir azalmaya sebep olduğu gözlendi. Sonuç olarak embriyolarda büyüme ve gelişmenin olumsuz yönde etkilendiği belirlendi.

Anahtar Kelimeler

• Anjiyogenezis,civciv,damar,koryoallantoik membran,sıcaklık

Kaynakça

1. Auger N, Naimi AI, Smargiassi A, et al. Extreme heat and risk of early delivery among preterm and term pregnancies. Epidemiology. 2014; 25:344-350.
2. Basu R, Chen H, Li DK, et al. The impact of maternal factors on the association between temperature and preterm delivery. Environ Res. 2017; 154:109-114.
3. Basu R, Malig B, Ostro B. High ambient temperature and the risk of preterm delivery. Am J Epidemiol. 2010; 172:1108-1117.
4. Bettencourt MC, Bauer JJ, Sesterhenn IA, Connelly RR, Moul JW. CD34 immunohistochemical assesment of angiogenesis as a prognostic marker for prostate cancer recurrence after radical prostatectomy. The Journal of Urology. 1998; 160:459-65.
5. Christensen VL, Donaldson WE, Nestor KE. Length of the plateau and pipping stages of incubation affects the physiology and survival of turkeys. Br Poult Sci. 1999; 40:297–303.
6. Costa C, Soares R, Schmitt F. Angiogenesis: now and then. APMIS. 2004; 112:402-412.
7. Dupertuis YV, Delie F, Cohen M, Pichard C. In ovo method for evaluating the effect of nutritional therapies on tumor development, growth and vascularization. Clinical Nutrition Experimental. 2015; 2:9-17.
8. Ertekin T, Bilir A, Aslan E, Koca B, Turamanlar O, Ertekin A, Albay S. The effect of diclofenac sodium on neural tube development in the early stage of chick embryos. Folia Morphol (Warsz). 2019; 78(2):307-313.

9. Folkman J. How is blood vessel growth regulated in normal and neoplastic tissue?G.H.A. Clowes memorial award lecture. Cancer Res. 1986; 46(2):467-73.
10. French NA. Effect of incubation temperature on the gross pathology of turkey embryos. Br Poult Sci. 1994; 35:363–371.
11. Guvenc Y, Billur D, Aydin S, Ozeren E, Demirci A, Alagoz F, Dalgic A, Belen D. Metamizole Sodium Induces Neural Tube Defects in a Chick Embryo Model. Turk Neurosurg. 2016; 26(3):445-8.
12. Guvenc Y, Dalgic A, Billur D, Karaoglu D, Aydin S, Daglioglu E, Ozdol C, Nacar OA, Yildirim AE, Belen D. The effects of levetiracetam on neural tube development in the early stage of chick embryos. Turk Neurosurg. 2013; 23(5):617-22.
13. Ha S, Liu D, Zhu Y, et al. Ambient temperature and early delivery of singleton pregnancies. Environ Health Perspect. 2017a; 125:453-459.
14. Ha S, Zhu Y, Liu D, et al. Ambient temperature and air quality in relation to small for gestational age and term low birthweight. Environ Res. 2017b; 55:394-400.
15. Hagedorn HG, Nerlich AG. Microvessel density and endothelial basement membrane composition in laryngeal squamous cell carcinomas. Acta Otolaryngol. 2000; 120:891-98.
16. Hamburger V, Hamilton HL. A series of normal stages in the development of the chick embryo. 1951. Dev Dyn. 1992; 195(4):231-72.
17. Hazel SJ. A novel early chorioallantoic membrane assay demonstrates quantitative and qualitative changes caused by antiangiogenic substances. J Lab Clin Med. 2003; 141:217-228.
18. Hulet R, Gladys G, Hill D, et al. Influence of egg shell embryonic incubation temperature and broiler breeder flock age on posthatch growth performance and carcass characteristics. Poult Sci. 2007; 86(2):408-412.
19. Kumar V, Abbas AK, Fausto N. Tissue renewal and repair: regeneration, healing, and fibrosis. In: Robbins and Cotran Pathologic Basis of Disease. 7th edition, Philedelphia, Pennsylvania. Elsevier Saunders Company. 2005; 87-119.
20. Leng T, Miller JM, Bilbao KV, et al. The chick chorioallantoic membrane as a model tissue for surgical retinal research and simulation. Retina. 2004 Jun; 24(3):427-434.
21. Leksrisompong N, Romero-Sanchez H, Plumstead PW, et al. Broiler incubation. 1. Effect of elevated temperature during late incubation on body weight and organs of chicks. Poult Sci. 2007; 86(12):2685-2691.
22. Logan A. Angiogenesis. Lancet. 1993; 341:1467-1468.
23. Lourens A, van den Brand H, Meijerhof R, et al. Effect of eggshell temperature during incubation on embryo development, hatchability, and posthatch development. Poult Sci. 2005; 84(6):914-920.
24. Lourens A, van den Brand H, Heetkamp MJ, et al. Effects of eggshell temperature and oxygen concentration on embryo growth and metabolism during incubation. Poult Sci. 2007; 86(10):2194-2199.
25. Maatjens CM, van Roovert-Reijrink IA, Engel B, et al. Temperature during the last week of incubation. I. Effects on hatching pattern and broiler chicken embryonic organ development. Poult Sci. 2016; 95(4):956-965.
26. Maatjens CM, Reijrink IA, Molenaar R, et al. Temperature and CO2 during the hatching phase. I. Effects on chick quality and organ development. Poult Sci. 2014; 93(3):645-654.
27. Maatjens CM, van Roovert-Reijrink IA, van den Anker I, et al. Temperature during the last week of incubation. II. Effects on first week broiler development and performance. Poult Sci. 2016; 95(9):2136-2144.
28. Mangir N, Raza A, Haycock JW, Chapple C, Macneil S. An improved in vivo methodology to visualise tumour induced changes in vasculature using the chick chorionic allantoic membrane assay. In Vivo. 2018; 32(3):461-472.
29. Meijerhof R. The influence of incubation on chick quality and broiler performance. Proceedings of the 20th Australian Poultry Science Symposium, 2009, Sydney, New South Wales, Australia, 9-11th February 2009. 2009; pp.167-170.
30. Molenaar R, Hulet R, Meijerhof R, et al. High eggshell temperatures during incubation decrease growth performance and increase the incidence of ascites in broiler chickens. Poult Sci. 2011; 90(3):624-632.
31. Nicholson B, Theodorescu D. Angiogenesis and prostate cancer tumor growth. J Cell Biochem. 2004; 91(1):125-150.
32. Norrby K. In-vivo models of angiogenesis. J Cell Mol Med. 2006; 10:588-612.
33. Oral M, Çakar S. Deneysel hayvan çalışmalarında etik prensipler. Anestezi Dergisi 2005; 13(2):75-82.
34. Özgürtaş Y. Anjiyogenezde bir in-vivo model: civciv koriyoallantoik membran. Gülhane Tıp Dergisi 2009; 51:67-69.
35. Peng CC, Hsieh CL, Lin LY, et al. Anisotropic diffusion deviates chicken embryo chorioallantoic membrane assay (CAM) to reflect inherent therapeutic behaviors. Colloids Surf B Biointerfaces. 2009; 70(1):29-34.
36. Ribatti D. The chick embryo chorioallantoic membrane as a model for tumor biology. Exp Cell Res. 2014 Nov 1;328(2):314-24.
37. Ribatti D. The chick embryo chorioallantoic membrane (CAM). A multifaceted experimental model. Mech Dev. 2016; 141:70-77.
38. Ricklefs RE. Comparative analysis of avian embryonic growth. J Exp Biol. 1987; 1:309–323.
39. Saraydaroğlu Ö, Özuysal S, Bilgin T. İnvaziv serviks karsinomlarında anjiogenezin prognostik faktörler üzerine etkisi. Türk Patoloji Dergisi. 2004; 20(1-2):14-17.
40. Strand LB, Barnett AG, Tong S. Maternal exposure to ambient temperature and the risks of preterm birth and stillbirth in Brisbane, Australia. Am J Epidemiol. 2012; 175(2):99-107.
41. Strohmeyer D, Rössing C, Bauerfeind A, Kaufman O, Sclhlechte H, et al. Vasculer endothelial growth factor and its correlation with angiogenesis and p53 expression in prostate cancer. Prostate. 2000; 45:216-224.
42. Townsend CM, Beauchamp DR, Evers MB, Mattox KL. Sabiston textbook of surgery: the biological basis of modern surgical practice. 17th ed. Elsevier Saunders, Philadelphia, 2004.
43. Zhang JJ, Jo JO, Huynh DL, Ghosh M, Kim M, et al. Lethality of inappropriate plasma exposure on chicken embryonic development. Oncotarget. 2017; 8(49):85642–85654.
44. Zhang Y, Yu C, Wang L. Temperature exposure during pregnancy and birth outcomes: an updated systematic review of epidemiological evidence. Environ Pollut. 2017; 225:700-712.