There are two types of visual pigments in fish eyes; most marine fishes have rhodopsin, while most freshwater fishes have porphyropsin. The biochemical basis for this dichotomy is the nature of the chromophores, retinal (A1) and 3-dehydroretinal (A2), each of which is bound by an opsin. In order to study the regional distribution of these visual pigments, we performed a new survey of the visual pigment chromophores in the eyes of many species of fish. Fish eyes from 164 species were used to examine their chromophores by high-performance liquid chromatography--44 species of freshwater fish, 20 of peripheral freshwater fish (coastal species), 10 of diadromous fish and 90 of seawater fish (marine species) were studied. The eyes of freshwater fish, limb freshwater fish and diadromous fish had both A1 and A2 chromophores, whereas those of marine fish possessed only A1 chromophores. Our results are similar to those of previous studies; however, we made a new finding that fish which live in freshwater possessed A1 if living near the sea and A2 if living far from the sea if they possessed only one type of chromophore.
It is well established that the zonae pellucidae of mature unfertilized eggs have the ability to induce the acrosome reaction of capacitated spermatozoa. To determine if this capacity of the zona is species-specific, hamster spermatozoa were allowed to attach to the zonae of homologous and heterologous eggs and examined for the acrosome reaction. The zonae of eggs from six different species were tested and the zona of hamster egg was found to have the strongest capacity to induce the acrosome reaction of hamster spermatozoa, followed by human and rat zonae. The zonae and mouse, guinea pig, and domestic fowl eggs were incapable of inducing the acrosome reaction of hamster spermatozoa. The acrosome reaction-inducing ability of the hamster zona was found to increase during maturation in the ovary. The zona of mature unfertilized hamster eggs maintained their acrosome reaction-inducing ability even after aldehyde fixation or storage in a highly concentrated solution of ammonium sulfate.
As gastrulation proceeds during sea urchin embryogenesis, primary mesenchyme cells (PMCs) fuse to form syncytial cables, within which calcium is deposited as CaCO 3 , and a pair of spicules is formed. Earlier studies suggested that calcium, previously sequestered by primary mesenchyme cells, is secreted and incorporated into growing spicules. We examined the effects of gadolinium ion (Gd 3+ ), a Ca 2+ channel blocker, on spicule formation. Gd 3+ did not lead to a retardation of embryogenesis prior to the initiation of gastrulation and did not inhibit the ingression of PMCs from the blastula wall or their migration along the inner blastocoel surface. However, when embryos were raised in seawater containing submicromolar to a few micromolar Gd 3+ , of which levels are considered to be insufficient to block Ca 2+ channels, a pair of triradiate spicules was formed asymmetrically. At 1-3 lmol ⁄ L Gd 3+ , many embryos formed only one spicule on either the left or right side, or embryos formed a very small second spicule. Induction of the spicule abnormality required the presence of Gd 3+ specifically during late blastula stage prior to spicule formation. An accumulation or adsorption of Gd
3+was not detected anywhere in the embryos by X-ray microanalysis, which suggests that Ca 2+ channels were not inhibited. These results suggest that Gd 3+ exerts an inhibitory effect on spicule formation through a mechanism that does not involve inhibition of Ca 2+ channels.
Before fertilization, capacitation and the acrosome reaction in mammalian spermatozoa must be completed. The motility and fertility of hamster sperm were examined in four kinds of modified Tyrode's solution with or without bovine serum albumin (BSA). Since the presence or absence of polyvinylalcohol (PVA) in the media was another variable, its effect on the sperm motility and fertility was also studied. Sperm were incubated in four different media for up to 6 h at 37.5 degrees C. After 4 h of incubation in the media containing BSA alone or BSA and PVA, sperm were hyperactivated, showing a high sperm motility index (SMI) and were able to fertilize more than 80% of eggs. However, their fertility rapidly decreased during further incubation. In contrast, sperm in the medium containing PVA and no BSA showed low SMI scores after 4 h. However, during the following 2-h period, the SMI progressively increased and sperm were hyperactivated. Furthermore, the hyperactivated sperm in the PVA containing medium were able to effectively fertilize eggs. Our results indicate that hamster sperm can be capacitated in BSA-free medium and that capacitation occurs much more slowly in such a medium. We suggest that PVA is a reasonable alternative to BSA in in vitro fertilization and that this slowly progressing system may be a good model for studying various steps in sperm activation.
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