Changes in the Localization of Immunoreactive Avian Beta-Defensin-8, -10 and -12 in Hen Ovarian Follicles during Follicular Growth

Abstract

Avian beta-defensins (avBDs) play significant roles in the innate immune system. The aim of this study was to identify immunoreactive (ir) avBDs proteins in the hen ovarian follicles and the changes in their localization with follicular growth. The ovarian follicles at di erent growth stages, namely the largest (F ), second and third largest (F and F ), prehierarchal small yellow and cortical follicles, were collected. The presence of ir-avBD-, -, and -were examined by immunohistochemistry and Western blot. The three ir-avBDs showed a similar pattern of immunostainings in the follicular tissues at di erent growth stages. In the granulosa cells, the ir-avBDs were identified in the cortical follicles, whereas their density was reduced in small yellow follicles. The granulosa cells of yellow follicles (F -F ) showed dense immunolabelings. The interstitial cells showed a faint immunolabeling for avBD-but not for avBD-and -in the cortical follicles, whereas they were weakly stained in the small yellow follicles. Dense immunoreaction products were noticed in the theca interna cells of F -F follicles. Western blot analysis showed a single band for each defensin. These results suggest that avBD-, -andproteins are expressed in the specific cells in the follicles, namely interstitial or theca interna cells and granulosa cells, where their amounts are likely increased with follicular growth. These avBDs may play significant roles in the host innate immune system in the follicles.

The comprehensive mechanisms underlying nonhierarchical follicular development in geese (Anser cygnoides)

“…The cell layers are primarily responsible for the transport of nutrients, mechanical support of growing oocytes, and the biosynthesis of steroids, which are important for regulating the rapid growth of follicles (Guraya, 1989;Wojtysiak et al, 2011;Rodler and Sinowatz, 2013). Recent data have further confirmed that the follicle wall secretes immunity peptides (Abdelsalam et al, 2010) and determines the fate of follicles, advancing in development or undergoing atresia (Quirk et al, 2004). The dynamic and highly regulated process of follicular development requires the coordinated actions of multiple genes via an orderly process orchestrated at the transcriptional and posttranscriptional levels (Johnson and Woods, 2009;Baley and Li, 2012;Shi et al, 2013).…”

Effects of Probiotics on the Expression and Localization of Avian β-defensins in the Proventriculus of Broiler Chicks

“…The anti-AvBD12 antibody had been raised in rabbit and purified using a Hi Trap affinity protein G HP column (GE Health Bio-Sciences AB, Uppsala, Sweden) and was used in our previous studies (Abdel Mageed et al, 2009;Abdelsalam et al, 2010). The tissues were fixed with 10% (vol/vol) formalin in phosphatebuffered saline (PBS), dehydrated, embedded in paraffin wax and sliced into sections of 4 μm thickness for histological and immunohistochemical examinations.…”

Induction of avian β-defensins by CpG oligodeoxynucleotides and proinflammatory cytokines in hen vaginal cells in vitro

“…The immunohistochemistry was performed using rabbit antibodies to AvBD3, -10, and -12 for primary antibodies that were used in our previous studies (Abdel Mageed et al 2009, Abdelsalam et al 2010. Vecta Stain ABC rabbit IgG kit (Vector Lab., Inc.) was used to identify the immunoreaction products.…”