Yücel Başımoğlu Koca

Genotoxic and histopathological effects of water pollution on two fish species, Barbus capito pectoralis and Chondrostoma nasus in the Büyük Menderes River, Turkey.

The genotoxic and histopathological effects of water pollution were investigated on two fish species caught from the Buyuk Menderes River and from its tributary, the Cine Stream. The Buyuk Menderes basin is an important agricultural area in Turkey. The levels of copper, zinc, cadmium, cobalt, and lead were measured at the surface of the water and in gills, liver, and muscle tissue of Chondrostoma nasus and Barbus capito pectoralis. In some tissues, the concentrations of some of these metals exceeded acceptable levels for human consumption. Zinc was found to be the most abundant metal in water and tissues. Maximal metal accumulation was observed in the liver. To detect the genotoxic potential of contaminants, the formation of micronucleus in erythrocytes was used as indicator of chromosomal damage. The frequency of micronucleus formation did not show significant differences between locations and controls in B. capito pectoralis caught from three locations and C. nasus from two locations. The histological changes included significant decreases of the mean lengths of primary and secondary lamellae. In gills epithelia, we observed cellular proliferation that developed Because of secondary lamellae fusion, ballooning degenerations, or club deformation of secondary lamellae and cystic structures in secondary lamellae. In the liver, the changes included swollen and ruptured parenchymal cells, loss of cord structure, vacuoles filled with cellular debris, focal necrosis, and a significant increase in Kupffer cells.

Immunohistochemical detection of insulin-like growth factor-I, transforming growth factor-β2, basic fibroblast growth factor and epidermal growth factor-receptor expression in developing rat ovary

The aim of this study was to determine the immunohistochemical expression and localization of insulin-like growth factor-I (IGF-I), transforming growth factor-beta2 (TGF-beta2), basic fibroblast growth factor (bFGF) and epidermal growth factor-receptor (EGF-R) in developing rat ovaries. Eighteen female Wistar rats were enrolled in this study; newborn (n=6), one-month-old (n=6) and adult (n=6) rats. Formalin-fixed and parafin-embedded ovarian tissues were stained with antibodies against IGF-I, TGF-beta2, bFGF and EGF-R, immunohistochemically. The ovarian cells were evaluated by semi-quantitative scoring system under light microscope. The staining of IGF-I, TGF-beta2, bFGF and EGF-R were most intense in the oocytes and were heavily at one-month-old rats. A moderate immunostaining in theca cells and corpus luteii reacted with IGF-I in adult rats. Furthermore the staining intensity for IGF-I was moderate in granulosa cells of newborn rat ovaries. We detected also a moderate staining for TGF-beta2 in corpus luteii of adult rats. In addition, we found a bFGF immunostaining mainly in oocytes of follicles of young and adult rats. Immunostaining for EGF-R was moderate in granulosa cells of one-month-old rats. In conclusion, this study suggests that growth factors play a pivotal role in ovarian function, especially in follicular development. The role of growth factor in controlling degeneration or growth (or both) of ovary follicles remain as explained.

Effects of Constant Lightness, Darkness and Parachlorophenylalanine Treatment on Tail Regeneration in the Lizard Ophisops elegans macrodactylus: Macroscopic, Biochemical and Histological Changes

A better understanding of regenerative growth is very important for the development of new potential strategies. Recently, the pineal secretory product melatonin was shown to stimulate the regeneration process. In this study, we carried out an experimental investigation of tail regeneration in young adult lizards, Ophisops elegans macrodactylus Berthold, 1842, addressing the role of melatonin on the regeneration process. Lizards were divided into three groups: constant light‐exposed control group (n = 13), constant dark‐exposed group (n = 15) and parachlorophenylalanine (p‐CPA) treatment group exposed to constant light (n = 15). Using a tail autotomy procedure, the effects of p‐CPA treatment on connective tissue together with neural tissue and vascular tissue in regenerating tail in lizards were investigated. p‐CPA (400 μg/kg body mass) was injected from day 0 to 30 days after autotomy. p‐CPA treatment produced a significant increase in the length of the regenerated tail compared with light‐exposed control and dark‐exposed lizards. Total collagen content was found to be higher in p‐CPA‐treated animals in comparison with other groups. Histologically, a higher percentage of connective tissue and vascular tissue and a lower percentage of neural tissue were found in the regenerated tails of the p‐CPA‐treated lizards. Importantly, the percentage of neural tissue in lizards in the dark‐exposed animals was higher than that in animals of both light‐exposed and p‐CPA treatment groups. Thus, it is clear that p‐CPA has a stimulatory influence on fibroblast collagen production and vascularization of the regenerated tail in the lizards. Furthermore, it seems that the neural regeneration process was markedly enhanced in lizards exposed to continuous darkness. Based on the results of our study, it is suggested that melatonin may be an active factor that speeds up the rate of tail regeneration in lacertilians.

Morphology, and Muscle- and Papilla-Volume Ratios, of the Tongue of Laudakia stellio (Agamidae, Squamata): A Histological and Stereological Study

Abstract We examined the histological structure of the tongue of Laudakia stellio, the starred agama lizard (Agamidae, Squamata), under light microscopy. We also investigated the muscle and papilla volume ratios, with volumes of each aspect of interest estimated according to the Cavalieri method. The macroscopically short, thick and muscle-rich front tip of the tongue of L. stellio does not show any bifurcation, and under light microscopy, the oval-shaped papilla-free front tip was seen to be covered by keratinized stratified epithelium. The dorsal and ventral parts were different, with the former partially covered by keratinized stratified epithelium and rich in secretory glands and secretory cells. The ventral part, which contained keratinized stratified cells, had a flat surface with no papillae. The dorsal surface of the anterior and posterior parts contained fungiform papillae, with the apical parts of these papillae containing minimal keratin; the interpapillar space was covered by keratin-free squamous stratified epithelium. The middle section of the tongue contained cylindrical-type papillae, with serous and mucous secretory glands and ducts at their base. Finally, the frontal and middle parts of the ventral and dorsal surfaces did not contain any taste buds, although there were some in the hind part of the dorsal surface. As morphometric estimates of volumes of the muscles and papillae, the mean volume ratios (relative to total tongue volume)±standard deviation were 0.66±0.03 and 0.33±0.03, with mean coefficients of error of 0.02 and 0.03, respectively.

Matrix changes due to the toxic effects of metronidazole in intestinal tissue of fish (Onchorhynchus mykiss)

Metranidazole (MTZ) is an antibiotic used for parasitic infections in a number of species. Accumulation of this drug in the environment and its interaction with fish of economic value makes this drug particularly important. In the present study, we examined the histopathological effects of MTZ on the intestinal tissue of Oncorhynchus mykiss. The fish in aquarium were exposed to MTZ at doses of 5, 10, 20 mg/L for 2, 4 and 8 days. At the end of the experiments, macroscopic pathology or death were not observed at these doses. Histochemical staining with Haematoxylene-Eosin, Periodic Acid Schiff and Gomori Trichrome showed, depending on increased dose and prolonged duration, areas of necrosis, edema, inflammation, small tears at the tips of the villi and excretion with heterogenic distribution of the Goblet cells. Moreover, changes in the connective tissue of the intestines due to toxicity of MTZ and decreases in immunostaining of matrix proteins such as laminin and collagen IV, especially in the epithelium were observed. Findings of the present study would be useful to demonstrate the adverse effects of MTZ use, emphasizing the importance of the effect on fish which could be very important public health.

Effects of Ca2+ channel blocker verapamil on tissue regeneration in a lizard tail autotomy model: a biochemical and histological study

Ca2+ ions have been reported to augment the activities of many cell types including cellular proliferation and tissue regeneration. Moreover, it is well known that verapamil is a L-type voltage-gated Ca2+ antagonist with important clinical implications. To evaluate the role of Ca2+ ions in the regeneration of tail in lizards, verapamil was used in vivo to modulate the activity of intracellular Ca2+ in a lizard tail autotomy model. A total of 35 adult lizards were divided into three groups: lightness control group ( n = 11), darkness group ( n = 11) and verapamil treatment group ( n = 13). The tails of adult lizards were amputated by pinching off the tail at the 15 th segment from the vent to induce tail regeneration. The first two groups served as untreated constant lightness and darkness groups as controls, but the remaining group received intraperitoneally 1 mg/kg of verapamil. Following autotomy, the length of regenerating tails was measured at 10, 15, 20, 25, and 30 days post-amputation. At the end of the study, the regenerating tails from animals from each group were removed for collagen assay procedure and histological examination. We found that verapamil produced a reduction in the length of the regenerated tail compared to untreated lightness group and the percentage of tail replaced in verapamil treatment group was lower than those in lightness control group. Total collagen contents were found to be higher in lightness control group in comparison with darkness and verapamil treatment groups. Accordingly, a quantitative stereological evaluation showed a higher percentage of neural tissue and a lower percentage of connective tissue, as well as vascular tissue, in the cross-sections of the regenerated tails taken from Ca2+ channel blocker verapamil-treated lizards, as compared to other groups. In conclusion, our results suggest that verapamil influences a variety of processes including fibroblast collagen production, neurogenesis, and angiogenesis during tail regeneration in lizard, possibly due to inhibition of intracellular Ca2+ ion by verapamil.

Morphological and histochemical investigations of esophagogastric tract of a lizard, Laudakia stellio (Agamidae, Linnaeus 1758)

Histological structures of esophagus and stomach tissue samples of Lacerta stellio have been studied, and glycosaminoglycan (GAG) distribution has been histochemically determined. Histologically, esophagus and stomach of L. stellio are composed of four layers: mucosa, submucosa, muscularis mucosae and serosa. Mucosa of esophagus is covered by simple columnar ciliated epithelium with many mucous secreting goblet cells and contains branched tubular glands.Stomach of L. stellio is composed of fundus (oral and aboral) and pylorus regions. Mucosa is covered by columnar epithelium. Fundic glands are branched tubular glands while pyloric glands are usually simple tubular glands. In both regions of the stomach, glands are subdivided into three areas as base, neck and isthmus. Both in the esophagus and stomach, muscular layer is in the form of smooth muscle having inner circular and outer longitudinal layers.According to the results obtained by Alcian Blue (pH 5.8)/Periodic Acid Schiff staining, stomach is similar to esophagus in that neutral mucins and hyaluronic acid (HA) are dominant in isthmus and neck regions of gland tissue of stomach. In the base of the stomach, only neutral mucins have been observed. HA has been observed to be dominant in all other regions of both stomach and esophagus, along with some but not much sulphated GAGs.

Histological structure of the skin of the Southern Crested Newt, Triturus karelinii (Salamandridae: Urodela)

Abstract The histological structure of the skin of the Southern Crested Newt, Triturus karelinii, from the active aquatic period has been studied by different staining methods. The species has two main layers: epidermis and dermis. The epidermal layer contains stratum germinativum and stratum corneum; the dermal layer stratum spongiosum and stratum compactum. The dermis has a connective tissue structure and contains large amounts of chromatophores and secretory glands. While the stratum compactum of the dermis contains collagen fibres, the basal laminae of the epidermal stratum germinativum cells, apical facies of the stratum corneum cells and the dermal ground substances contain glycosaminoglycan structures. By using computerized image processing software, the thickness of the epidermis and dermis has been measured and the amount of collagen fibres in terms of chosen point determined. We conclude that the extracellular matrix of the T. karelinii skin is largely comprised of two structurally distinct molecular components: collagen and glycosaminoglycans, but the amount of these components may be different in the water and land periods, depending upon the activity of these amphibians.