The effects of titanium dioxide nanoparticles on ultrastructure of zebrafish testis (Danio rerio).

INTRODUCTION: Nanotechnology investigates materials at nanoscale level (0.1-100nm in diameter). There are many commercially nanoproducts such as silver, silicon, titanium, zinc, and gold. They are used in a variety of applications and released to the environment. Titanium dioxide (TiO2) is one of the most commonly used nanoparticles (NP). In this study, the ultrastructural effects of TiO2-NP on zebrafish testis tissue were evaluated. MATERIAL AND METHOD: Zebrafish were divided into four groups (N=60) as one control and 3 experimental groups (1mg/L, 2mg/L and 4mg/L TiO2). Testis tissues were dissected after 5days of the exposure. Tissues were fixed with 2.5% glutaraldehyde at 4°C. After routine electron microscopy tissue processing, the testis were embedded in epon resin. Ultrathin sections were counterstained with 1% uranyl acetate and lead citrate and examined using a transmission electron microscope. RESULTS: Mitochodrial degeneration with swelling and cristae loss were detected in Sertoli cells and spermatogonial cells of TiO2-NP treated groups in a dose-dependent manner. Moreover, autophagic vacuole accumulation were seen in Sertoli cell cytoplasms of the experimental groups. Necrosis was also detected in the 4mg TiO2-NP-treated group. CONCLUSION: TiO2-NP has been used in crop production, food additives, medicine, toothpastes, sunscreens, cosmetics, and in waste water treatment, which contaminated the environment. Our findings showed TiO2-NP-induced autophagy and necrosis at higher doses in Sertoli cells, which consequently negatively affected spermatogenic cells and testicular morphology of Zebrafish. It is important to give much more attention to the use of this NP to minimise the possible effects on nature and organisms.

Developmental Toxicity of (4S)-2- (4-hydroxy-3-methoxyphenyl) thiazolidine-4-carboxylic acid in Zebrafish ( Danio rerio )

ABSTRACT (4S)-2-(4-hydroxy-3-methoxyphenyl)thiazolidine-4-carboxylic acid is new synthesized substance obtained from cysteine and valine. Thiazolidine derivates have important biological responses so scientists work intensively on these compounds in recent years. It is obvious that thiazolidine contained compounds will be used in future in the pharmaceutical industry to treat important diseases. Median lethal concentrations (LC50) for 48 h and 96 h were found as 1.106±0.052 mM and 0.804mM ± 0.102 respectively. According to LC50, exposure doses were determined as control, 0.4 mM, 0.2 mM and 0.1 mM (4S)-2-(4-hydroxy-3-methoxyphenyl)thiazolidine-4-carboxylic acid. Developmental toxicity and apoptotic features on zebrafish development were evaluated in this study. The results of this study indicate that (4S)-2-(4-hydroxy-3-methoxyphenyl)thiazolidine-4-carboxylic acid exposure cause developmental defects like pericardial edema, bent spine, tail malformation, blood accumulation, yolk sac edema but on the other hand concentration-dependent decrease in apoptotic rate. Likewise, concentration-dependent decrease in hatching and increase in mortality of embryos were also detected.

Ovarian follicle structure of Zebrafish (Danio rerio Hamilton) after poly(2-ethyl-2-oxazoline) exposure.

Poly(2-oxazoline)s have been widely known for their biomedical applications. They mimic natural systems and are generally used as liposomes, drug and gene delivery modules and also as pseudopeptides. In this study, effects of different doses (10 and 50 mg/L) of poly(2-ethyl-2-oxazoline) on ovarian follicle ultrastructure of zebrafish were investigated with light and electron microscopy. After five days of administation, ovary tissues were disected and routine histological processes were done. Severe structural deformation at zona radiata, follicular epithelium and outer granulosa cell structure were observed. Edema at follicular epithelium, zona radiata and vitelline envelope stuructures were detected. Dose dependent increase in the number of immature oocytes were seen. Briefly, it was demonstrated that poly(2-ethyl-2-oxazoline) exposure affected oogenesis and caused apoptosis in zebrafish.

Identification of Primordial Germ Cells: Cytological, Histological and Immunohistochemical Aspects

Primordial germ cells (PGCs) constitute an embryonic cell type that migrate to gonadal precursors and form the gametes. In many animals, PGCs are set apart from somatic cells early during embryogenesis. These cells migrate to gonadal precursors and then constitute gonads so they are useful models for cell motility studies. They have a highlighted importance for development and reproduction studies. Primordial germ cells have morphological differences from the somatic cells. Structure of these cells can be detected with light and electron microscopy in early development stages. This review describes the morphological, histological, molecular and ultrastructural features of primordial germ cells in different animals and gives an overview for simplified identification.

The effects of permethrin on rat ovarian tissue morphology.

All organisms are exposed to chemical agents during their lifetime. One of these agents is a pesticide that is used as fly killer. In this study we investigated the effects of permethrin on rat ovaries using light and electron microscopy. We used 24 Wistar albino female rats and divided them into 3 groups. Dosages 20 and 40 mg/kg/day permethrin were administered by gavage for 14 days. Normal saline was given to control rats. After treatment, ovarian tissues were collected and prepared for light and electron microscopy evaluation. Negative effects of permethrin were detected on follicular and corpus luteum cell morphology in a dose dependent manner when compared with the control group. Picnotic cellular appearance and condensed chromatin were detected as evidence of apoptotic cell death. Furthermore, degenerative changes were seen in the ultrastructure of mitochondria and endoplasmic reticulum. Thus, these findings suggested that permethrin caused degenerative effects on ovarian morphology in a dose dependent manner.

Ovary maturatýon stages and histological investigation of ovary of the Zebrafish (Danio rerio)

Microscopic ovary features of Zebrafish were analyzed. Samples were fixed in Bouin’s solution, mounted in parafin and cut into 6-7 µm-thick slices which were stained with Hematoxylin and Eosin (HE), methylene blue and using Periodic-acid Schiff method (PAS). The thin sections in ultramicrotome with the thickness of approximately 0,5 - 0,7 µm were examined by using Jeol electron microscope following contrasting with Uranyl Acetate-Lead Citrate. The zebrafish was an extremely dynamic organ in which follicles underwent asynchronous development. The oocytes of Zebrafish ovary were observed in various phases. The oocyte development of zebrafish was divided into four stages (primary growth, cortical alveolus, vitellogenic and mature oocyte). Oocyte diameters were observed to vary between 0.,08 mm and 0.76 mm.