The development of rat Leydig cell progenitors in vitro: how essential is luteinising hormone?

Luteinising hormone(LH) appears to be important for the establishment of the adult-type Leydig cell population. The role of LH in the initial steps of stem Leydig cell/precursor cell differentiation is less clear. The aim of the present study was to elucidate the role of LH in the differentiation of spindle-shaped mesenchymal-like cells into Leydig cell progenitors. Interstitial cells were isolated from rat testes at three different ages reflecting different phases in development, and cultured in the presence of increasing concentrations of LH (ranging from 0.01 to 10 ng/ml). Cells were isolated from 10-day-old rats when stem Leydig cells/precursor cells are abundant; 13-day-old rats when the first 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD)-positive Leydig cell progenitors have developed in the strain of rats used in this study; and 18-day-old rats just prior to the wave of progenitor proliferation. Immunohistochemistry revealed that before stem Leydig cells differentiate into progenitor cells, they acquire functional LH receptors and become precursor cells. This was confirmed by an in vivo immunohistochemical double-labelling experiment. Addition of LH to the cultures increased the percentage of LH/3 beta-HSD-labelled Leydig cell progenitors, while the percentage of cells solely expressing the LH receptor decreased. Cell proliferation was negligible, suggesting that the increase in 3 beta-HSD-positive cells is the result of precursor cell differentiation. When interstitial cells were isolated from 13-day-old rats and to a lesser extent from 10-day old rats, a small proportion of the precursors could develop into progenitor cells independent of the presence of LH. In conclusion: before Leydig stem cells differentiate into 3 beta-HSD-positive progenitor cells, they acquire LH receptors and become precursor cells. LH appears to be essential, even at very low doses for the differentiation of these precursor cells into 3 beta-HSD-positive progenitors, although a subpopulation of precursor cells can develop into progenitors independently of LH

The ‘Coral Bulker’ Fuel Oil Spill on the North Coast of Portugal: Spatial and Temporal Biomarker Responses in Mytilus galloprovincialis

In December 2000, the ship ‘Coral Bulker’ ran aground at the entrance of the port of Viana do Castelo (North–west coast of Portugal). A large amount of fuel oil was spilled and part of it reached the shore. To evaluate the spatial and temporal impact of this oil spill, a field study, and several laboratory toxicity tests were performed using Mytilus galloprovincialis as biological indicator of environmental contamination and the biomarkers glutathione S-transferases (GSTs) and acetylcholinesterase (AChE) as indicative criteria. Fifteen days after the oil spill, mussels collected at stations located near the ship presented higher and lower values of GSTs and AChE activity, respectively. These results, and those obtained in the laboratory toxicity tests, evidence that these biomarkers were sensitive indicators of exposure to this kind of pollution and were able to monitor a spatial impact of the oil spill of at least 10 km, confirming the higher level of contamination near the ship and a contamination gradient along the sampling stations. One year after the accident, such a contamination gradient was no longer evident. This study highlight the potential suitability of a biomarker approach for assessing spatial and temporal impacts of marine pollution accidents, such as fuel oil spills, suggesting the inclusion of these biomarkers in risk assessment studies, as cost-effective and early warning recognized tools. Major advantages and limitations of the biomarker approach used in this study are further discussed