The authors have previously demonstrated that Streptococcus mutans shows an exponential-phase acid-tolerance response following an acid shock from pH 75 to 55 that enhances survival at pH 30. In this study the response of S. mutans H7 to acid shock was compared with the responses generated by salt, heat, oxidation and starvation. Prior induction of the acid-tolerance response did not cross-protect the cells from a subsequent challenge by the other stresses ; however, prior adaptation to the other stresses, except heat (42 mC), protected the cells during a subsequent acid challenge at pH 35. Starvation by fivefold dilution of the basal medium (BM) plus fivefold reduction of its glucose content increased the numbers of survivors 12-fold, whereas elimination of glucose from fivefold-diluted BM led to a sevenfold enhancement compared to the control cells ; this indicated a relationship between the acid and starvation responses. The stress responses were further characterized by comparing the 2D electrophoretic protein profiles of exponential-phase cells subjected to the various stress conditions. Cells were grown to exponential phase at pH 75 (37 mC) and then incubated for 30 min under the various stress conditions in the presence of 14 C-labelled amino acids followed by cell extraction, protein separation by 2D gel electrophoresis and image analysis of the resulting autoradiograms. Using consistent twofold or greater changes in IOD % as a measure, oxidative stress resulted in the upregulation of 69 proteins, 15 of which were oxidation-specific, and in the downregulation of 24 proteins, when compared to the control cells. An acid shock from pH 75 to 55 enhanced synthesis of 64 proteins, 25 of them acidspecific, while 49 proteins exhibited diminished synthesis. The dilution of BM resulted in the increased formation of 58 proteins, with 11 starvation-specific proteins and 20 showing decreased synthesis. Some 52 and 40 proteins were enhanced by salt and heat stress, with 10 and 6 of these proteins, respectively, specific to the stress. The synthesis of a significant number of proteins was increased by more than one, but not all stress conditions ; six proteins were enhanced by all five stress conditions and could be classified as general stress proteins. Clearly, the response of S. mutans to adverse environmental conditons results in complex and diverse alterations in protein synthesis to further cell survival.

One of the reasons that the crystalline lenses of vertebrate eyes are highly transparent is that most of the cells have broken down all of their organelles, including the nuclei. These cells can neither synthesize new proteins nor generate energy by electron transport in the mitochondria. Only in the peripheral layers--in the cichlid fish Aequidens pulcher, beyond 92% of the lens radius--are there cells with full complements of organelles. We report here that the optical properties of the lens change between the light-adapted and dark-adapted states in A. pulcher. Changes occur even in cell layers free of organelles, and they occur in parallel with changes in retinal function between the light-adapted (all-cone, color vision) and dark-adapted (all-rod, grayscale vision) states. Depleting the eye of dopamine also caused changes in the optical properties similar to those of dark adaptation. Our results indicate that the refractive index of the organelle-free lens fiber cells can be adjusted quickly and accurately.

No abstract

The crystalline lens in the cichlid fish Aequidens pulcher undergoes a transformation of its optical properties every dawn and dusk as the eye adapts to changes in light conditions. During dusk the transformation result in an increase of the refractive power in the lens cortex, the outermost 40 percent. The change is thought to match the optical properties of the lens to the requirements of the retina. Using a short term in vitro lens culturing system together with optical measurements we here present data that confirm that the optical properties of the lens can change within hours and that dopamine influences the optical properties of the lens. Dopamine yields dose-dependent decrease of the refractive power in the lens cortex. The D1-agonist SKF-38393 induces a similar decrease of the refractive power in the cortex, while the D2-agonist quinpirole has no effect. The effect of dopamine can be blocked by using the D1-antagonist SCH 23390. Our results suggest that dopamine alone could be responsible for the light/dark adaptive optical changes in the lens, but the involvement of other signaling substances cannot be ruled out.

Culturing whole lenses is a frequently used method for studying regulatory events on the lens in controlled environments. The evaluation methods used often fall under two categories, molecular or optical. The main benefit from optical measurements is that they directly detect changes in the lens' main function, i.e. refracting light. However, these measurements often have rather low resolution or yield results open for subjective interpretation. Here we present a short-term crystalline lens culturing technique combined with a high-resolution optical measuring method. There are two main advantages of using teleost lenses compared to mammalian lenses. Teleost tissue generally has a higher tolerance than mammalian tissue with regard to temperature and nutrient fluctuations. Teleost lenses are structurally more robust and can be excised from the eye without disturbing form or function. The technique is developed for short-term culturing (3 h), however, the lenses appear viable for at least 24 h and longer culturing may be possible. The technique is resistant to small variations in osmolarity and yields quantitative datasets for further analyses and statistical treatment.