İpekböceği, Bombyx mori L., 1758 (Lepidoptera: Bombycidae) orta bağırsak kök hücrelerinde ekdizonla ilişkili genlerin ekspresyon profilleri ve olası rolleri

Böcek orta bağırsağı, özellikle kök hücreler tarafından hücre düzenlemesinin kontrol edilmesiyle ilgili olarak omurgalı bağırsağıyla dikkate değer benzerliklere sahiptir. Pupa orta bağırsağı, kök hücreler tarafından oluşturulur ve ekdizon, orta bağırsağın yeniden şekillenme olaylarını düzenlerken, juvenil hormon, ekdizon salınımını baskılayarak engeller. Bu çalışma, larva-pupa metamorfozu sırasında Bombyx mori L., 1758 (Lepidoptera: Bombycidae)'nin kök hücrelerinde ekdizonla ilişkili genlerin olası işlevlerini araştırmıştır. Bu çalışma 2014-2018 yılları arasında Ege Üniversitesi ipek böceği kültür laboratuvarı ve böcek fizyolojisi araştırma laboratuvarında yapılmıştır. Juvenil hormon analoğu fenoksikarb, pupa orta bağırsak oluşumunu geciktirmek veya engellemek için 5. dönem Bombyx larvalarına uygulanmıştır. Morfolojik gözlemler hematoksilen ve eozin boyama ile gerçekleştirilmiş; kök hücrelerin proliferasyon hızı, bromodeoksiüridin hücre proliferasyon analizi ile analiz edilmiş ve ekdizon ile ilgili genlerin ekspresyon durumları kantitatif gerçek zamanlı ters transkriptaz-polimeraz zincir reaksiyonu ile tespit edilmiştir. Genlerin ekspresyon durumları ve orta bağırsaktaki gelişimsel olaylar, uygulama zamanına bağlı olarak fenoksikarb tedavisinden farklı şekilde etkilenmiştir. Sonuçlara göre, genler önce fenoksikarb duyarlılıklarına göre sınıflandırılmış, daha sonra kök hücrelerin morfolojik değerlendirmeleri ile bağlantılı olarak ekspresyon profillerine göre gruplandırılmıştır.

Anahtar Kelimeler:

Bombyx mori, ekdizon bağlantılı genler, orta bağırsak, kök hücre

Expression profiles and possible functions of the ecdysone-related genes in the midgut stem cells of the silkworm, Bombyx mori L., 1758 (Lepidoptera: Bombycidae)

The insect midgut has remarkable similarities with the vertebrate intestine especially concerning controlling cell regulation by the stem cells. While the formation of the pupal midgut from stem cells is regulated by ecdysone, it is inhibited by juvenile hormone via suppression of ecdysone release. This study investigated the possible functions of ecdysone-related genes in the stem cells of Bombyx mori L., 1758 (Lepidoptera: Bombycidae) during larval-pupal metamorphosis. The study was conducted in the Ege University silkworm culture laboratory and insect physiology research laboratory during the years between 2014-2018. Juvenile hormone analogue, fenoxycarb was applied to the fifth instar Bombyx larvae to delayed or inhibit the formation of the pupal midgut. Morphologic observations were performed by hematoxylin plus eosin staining; the proliferation rate of stem cells was analyzed by bromodeoxyuridine cell proliferation assay and expression patterns of ecdysone-related genes were detected by quantitative real-time reverse transcriptase-polymerase chain reaction. Expression states of genes and developmental events of the midgut were differently affected by fenoxycarb treatment in an application time-dependent manner. According to results, genes were firstly classified based on their sensitivity of fenoxycarb, then grouped according to their expression profile in connection with morphological evaluations of stem cells.

Keywords:

Bombyx mori, ecdysone-related genes, midgut, stem cell,

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