Epidermal growth factor receptors and their ligands are expressed in the spleen of the Japanese Quail (Coturnix coturnix japonica) during the post-hatch period.

1. The epidermal growth factor (EGF) family plays an important role in the development, differentiation, migration and apoptosis of cells, as well as in wound healing, which are all essential to the viability of multicellular organisms. The avian spleen is a principal organ of systemic immunity and its importance in disease resistance is presumably accentuated by the scarcity of avian lymph nodes.2. The aim of this study was to determine whether EGF receptors (ErbB1-4) and their ligands (EGF, AREG and NRG) are expressed in the structural components of the quail spleen during the post-hatch period. At each selected age, from 1 d to 7, 14, 21 and 60 d, 10 quails were euthanised under ether anaesthesia and their spleens were fixed in a 10% formaldehyde-alcohol solution. Following routine histological processing, the streptavidin-biotin-peroxidase method was used for immunohistochemical examination.3. Strong cytoplasmic immunoreactions for ErbB2, ErbB4 and NRG were observed in the ellipsoid associated cells (EAC) of the quail spleen throughout the post-hatch period. This immunoreactivity in the EAC increased after the 7th d post-hatch. ErbB1 and ErbB3 immunoreactions were relatively similar and weak in all components of the spleen during the post-hatch period. Some immune cells of the peri-arterial lymphatic sheath (PALS) and peri-ellipsoidal lymphatic sheath (PELS) showed positive immunoreactivity for the ErbB receptors and their ligands. In the vascular smooth muscle cells, immunoreactivity for ErbB2 was stronger than that for the other ErbB receptors and their ligands.4. The data showed that ErbB receptors and their ligands (EGF, AREG and NRG) are expressed by different structural components of the quail spleen during the post-hatch period.

Distribution of CD68-, CD8-, MHCI- and MHCII-positive cells in the bull and ram testis and epididymis.

The mammalian testis possesses a special immunological environment because of its properties of remarkable immune privilege and effective local innate immunity. The testicular immune privilege protects immunogenic germ cells from systemic immune attack, and local innate immunity is important in preventing testicular microbial infections. Thus, this study aimed to immunohistochemically demonstrate the distribution and localization of CD68-, CD8-, MHCI- and MHCII-positive immune cells in the testes and epididymes. Negative immunoreactivity was detected in the seminiferous tubule epithelium and peritubular myoid cells of the testes upon staining in CD68, CD8 and MHC Class I. Positive CD68 immunoreaction was determined in the Sertoli cells and some Leydig cells. The detection of positive cells for CD8 clearly indicated the presence of lymphocytes. Furthermore, the staining with MHCI intensity was ascertained to vary from weak to moderate in the Sertoli and Leydig cells and connective tissue cells. MHCII-positive immunoreactivity was determined in myoid cells and Leydig cells in the interstitial area. The epithelium of the epididymis showed positive staining for CD68 and CD8, but the stroma displayed a rather weak staining. In the ram epididymis, neither intraepithelial nor interstitial positive reaction was observed for MHCI. In the epididymis, the basal cells displayed a stronger staining for MHCII. In conclusion, these cells not only contribute to local immunity through their direct effects on the quality of fertility in males, but also contribute either directly or indirectly to immune privilege by minimizing the development of both autoimmune reactions and potentially harmful risks.

Topical dimethyl sulfoxide inhibits corneal neovascularization and stimulates corneal repair in rabbits following acid burn.

Neovascularization of the cornea is characterized by the growth of blood vessels caused by imbalances between angiogenic and anti-angiogenic factors. We investigated whether the expression of Vascular endothelial growth factor (VEGF), Vascular endothelial growth factor receptor (VEGF), Vascular endothelial growth inhibitor (VEGI) receptors, as well as topical drug treatments, participate in regulating corneal neovascularization after corneal damage and remodeling. We used 72 mature male New Zealand rabbits. Corneal burns were induced by hydrofluoric acid under general anesthesia. The rabbits then were treated with indomethacin or dimethyl sulfoxide (DMSO). The animals were euthanized on days 2, 7 and 14 after injury. Each cornea was fixed with 10% neutral formalin. On days 2, 7 and 14, VEGF, flk1/KDR and flt1/fms were strongly expressed in the epithelial, stromal and inflammatory cells, but not in the corneal endothelial cells. On day 7, newly formed blood vessels were observed growing toward the center of the cornea. In the control, indomethacin treated, DMSO-treated, and indomethacin + DMSO-treated animals, VEGI, VEGF, and the receptors, flk1/KDR, flt1/fms and flt4, were expressed at different densities in the neovascular regions. This was particularly evident in the indomethacin- and indomethacin + DMSO-treated groups on days 7 and 14, compared to day 2. Treatment with VEGF and DMSO stimulated repair of corneal damage. We suggest that VEGI in the endothelial cells of neovascularized cornea may act as a signaling protein that promotes balance between cell proliferation and apoptosis. Topical administration of DMSO inhibited corneal neovascularization more effectively than indomethacin.

Expression of vascular endothelial growth factor receptors and their ligands in rat uterus during the postpartum involution period.

Vascular endothelial growth factor (VEGF) and its specific receptors, FLt1/fms, Flk1/KDR and FLt4, play important roles in vasculogenesis, and physiological and pathological angiogenesis. Whether angiogenic growth factors are involved in regulating angiogenic processes during the postpartum involution period (PP) of the rat uterus is unknown. We used immunohistochemistry to analyze the expression levels of VEGF, the fms-like tyrosine kinase 1 (FLt1/fms), the kinase insert domain-containing region 1 (Flk1/KDR), Fms-related tyrosine kinase 4 (FLt4) and vascular endothelial growth inhibitor (VEGI) in the rat uterus during the days 1, 3, 5, 10 and 15 of the PP to determine the temporal and spatial expressions of VEGF and its receptors during the PP. Throughout the PP, cytoplasmic and membrane staining of VEGI, VEGF and their receptors were observed in the lumens, crypts and glandular epithelial cells as well as in connective tissue and vascular endothelial and smooth muscle cells in the endometrium. We found that the intensity of the immunoreactions in the endometrium varied with the morphological changes that occurred during involution. Immunoreactions for VEGI, VEGF and their receptor, Flk1/KDR, in the luminal epithelial cells were stronger than those in the glandular epithelial and stromal cells, particularly during PP 1, 3 and 5, which suggests that these peptides may contribute to re-epithelialization of the endometrium. On the other hand, Flt1/fms immunoreactivity was strong mainly in the stromal cells during the PP. The presence of VEGF and its receptors (FLt1/fms, Flk1/KDR, FLt4) in the stromal cells and blood vessels during the PP suggests that they may contribute to regulating stromal repair and angiogenesis in the involuting uterus of the rat.

Anatomical and histological structure of the tongue and histochemical characteristics of the lingual salivary glands in the Chukar partridge (Alectoris chukar, Gray 1830).

1. The aim of the study was to examine the morphology of the tongue and the histochemical features of the lingual salivary glands in this species. 2. The tongue was elongated, terminating in a rather sharp, dagger-like apex. On the surface of the tongue and situated between the body and root of the tongue, two rows of conical papillae, the sharp apices of which pointed towards the posterior part of the tongue, were observed. The keratinised epithelium lining the dorsal surface lacked typical gustatory papillae. However, it was observed that taste buds were present in the epithelium of the lingual body and root. The tongue was supported by a structure composed of hyaline cartilage, the paraglossum, which extended from the lingual root to the apex. Simple branched tubular glands, which were encapsulated by connective tissue, were embedded within the submucosa in the body (anterior salivary glands) and root (posterior salivary glands) of the tongue. It was observed that the secretion of the lingual glands contained neutral mucins, proteoglycans containing carboxylic acid, weak and strong sulphated groups, N-acetylated sialomucins, but lacked glycogen. 3. It was demonstrated that, the general morphological features, papillary distribution of the tongue and the histological structure of the mucosa epithelium and the supportive elements displayed similarity to those of other domestic avian species. It was also determined that, in view of the particular feeding types, in the partridge, the presence of the papillary crest was not correlated with diet.

Expression of the erbB/HER receptor family in the bovine uterus during the sexual cycle and the relation of this family to serum sex steroids.

Our study was designed to investigate immunohistochemically the expression of the receptors of the erbB/HER family (erbB1/HER1, erbB2/HER2, erbB3/HER3, erbB4/HER4) in the bovine uterus during the follicular and luteal phases of the sexual cycle, and the relation to ovarian sex steroids. The stage of the estrous cycle in 30 Holstein bovine was assessed based on the gross and histological appearance of the ovaries and uterus, and on blood steroid hormone levels. Tissue samples taken from the uterus were fixed in 10% formaldehyde for routine histological processing. Positive membrane and cytoplasmic staining of varying intensity were determined in the uterus during the follicular and luteal phases of the sexual cycle for erbB/HER receptors in luminal and glandular epithelial cells, connective tissue, smooth muscle and vascular endothelial and smooth muscle cells. We demonstrated that the apical and basal membranes of luminal epithelial cells and the apical membrane of glandular epithelial cells reacted with erbB1/HER1 and erbB2/HER2 during both the follicular and luteal phases. The reaction for erbB3/HER3 and erbB4/HER4 was stronger in the cytoplasm of luminal and glandular epithelial cells, but was heterogeneous. During both the follicular and luteal phases, the percentage and staining intensity of luminal and superficial glandular epithelial cells reacting positively with the receptors erbB1/HER1, erbB2/HER2, erbB3/HER3 and erbB4/HER4 were greater than those of deep glandular epithelial and connective tissue cells (p < 0.05). We demonstrated that the expression of the erbB/HER receptor family varied with different cell types in the bovine uterus during the follicular and luteal phases.

The expression of vascular endothelial growth factor and its receptors (flt1/fms, flk1/KDR, flt4) and vascular endothelial growth inhibitor in the bovine uterus during the sexual cycle and their correlation with serum sex steroids.

The present study was conducted to demonstrate of the immunohistochemical localization of vascular endothelial growth factor (VEGF) and its receptors (flt1/fms, flk1/KDR and flt4) as well as vascular endothelial growth inhibitor (VEGI) and to determine the correlation of VEGF and its receptors and VEGI with serum sex steroids (estrogen and progesterone) in the bovine uterus during the sexual cycle. The stage of the estrous cycle in 30 Holstein cattle was assessed based on the gross and histological appearance of the ovaries and uterus and on blood steroid hormone levels. Tissue samples obtained from the uterus were fixed in 10% formaldehyde for routine histological processing. During both follicular and luteal phases, positive cytoplasmic and membrane staining was achieved for VEGF and its receptors (flt1/fms, flk1/KDR and flt4) as well as VEGI in the luminal and glandular epithelial cells, the connective tissue and smooth muscle cells, and the vascular endothelial cells and smooth muscle cells in the uterus. The intensity, proportional and total scores determined for VEGF and its receptors (flt1/fms and flt4) as well as VEGI were greater in the luminal and glandular epithelial cells compared to the connective tissue and smooth muscle cells (P < 0.05). Furthermore, the number and intensity of the flk1/KDR positive cells were greater among the connective tissue cells compared to the luminal and glandular epithelial cells (P < 0.05). As a result, it was determined that the expression of VEGF and its receptors as well as VEGI in the bovine uterus during the follicular and luteal phases varied with different cell types. This suggests that depending on the stage of the sexual cycle, these factors may mediate the establishment of an appropriate environment for the nutritional supply and implantation of the embryo primarily due to the stimulation of angiogenesis but also through the increase in the secretory activity of the epithelial cells in the uterus. Furthermore, this indicates that ovarian steroid hormones play a significant role in regulating the expression of VEGF and its receptors as well as VEGI.

Localization of estrogen receptor α and progesterone receptor B in bovine cervix and vagina during the follicular and luteal phases of the sexual cycle.

The localization and distribution of estrogen receptors (ERα) and progesterone receptors (PR-B) in the cervix and vagina of sexually mature bovines during the follicular and luteal phases of the sexual cycle were studied using immunohistochemistry. The estrous cycle stage of 23 Holstein bovines was assessed by gross and histological appearance of ovaries and blood steroid hormone values. Tissue samples from cervix and vagina were fixed in 10% formaldehyde for routine histological processing. Nuclear staining for ERα and PR-B was observed in the epithelial cells of the surface epithelium, stromal cells and smooth muscle cells. Generally, in the cervix, ERα immunoreactivity was more intense in the epithelial and smooth muscle cells during the follicular phase and in the epithelial cells during the luteal phase (p < 0.05). PR-B immunoreactivity was more intense in the epithelial and smooth muscle cells than in the superficial and deep stromal cells during the follicular and luteal phases (p < 0.05). In the vagina, ERα and PR-B immunoreactivities were more intense in the epithelial cells than in the connective tissue cells and smooth muscle cells during the follicular and luteal phases (p < 0.05). These results indicated that the frequency and intensity of ERα and PR-B immunoreactivity in the cervix and vagina of bovines varied according to the cervical and vaginal cell types and the phases of the sexual cycle.

Structure of the oesophagus and morphometric, histochemical-immunohistochemical profiles of the oesophageal gland during the post-hatching period of Japanese quails (Coturnix coturnix japonica).

In the oesophagus, mucins, which originate from oesophageal submucosal glands, play an important role in the mucosal protection as a pre-epithelial barrier. In this study, the structure of cervical and thoracic parts of oesophagus of Japanese quail during the post-hatching period was compared, and the contents of carbohydrate and gastric mucin MUC5AC of the oesophageal glands in these parts were analysed at the light microscope levels by applying conventional histochemical and immunohistochemical methods. The oesophageal glands were present at hatching, located in the laminae propria. The numbers of glands were different in the cervical and thoracic parts, but the differences were found to be insignificant. The thoracic part has the oesophageal tonsils which are associated with the glands. Oesophageal tonsil was formed from day 5 after hatching. In quail of all ages, the secretory epithelium of glands contained neutral sialomucins and weakly sulphomucins. The cells in the neck region of secretory units contained sialomucins, while the cells of excretory ducts had strongly sulphomucins. Sialomucin containing cells in the secretory units increased with the advance of age and glandular development. But, in the secretory units, the sulphomucin content of glands was more in the thoracic part. The secretory epithelium of tonsil-associated glands contained mostly sulphomucins and a little sialomucin. From the hatching, MUC5AC mucin was detected in the cells of excretory ducts. Although the lymphoepithelium of the tonsil units exhibited negative reactions to all histochemical methods, it showed positive reaction to MUC5AC mucin antibody. In conclusion, the cervical and thoracic parts may be functionally different and the thoracic part of oesophagus was transformed into an immunological organ following day 5 after hatching.