A Two-Dimensional Electrophoresis Study of Phosphorylation and Dephosphorylation of Chromaffin Cell Proteins in Response to a Secretory Stimulus

Phosphorylated proteins of bovine chromaffin cells, radioactively labeled with [32P]orthophosphate, have been analyzed by two-dimensional polyacrylamide gel electrophoresis and autoradiography. Complex two-dimensional electrophoretograms were studied with the aid of computer-assisted image analysis (CAIA). A database map of 32P-labeled proteins was constructed; approximately 500 polypeptides have been detected, numbered, and characterized according to the intensity of labeling, molecular weight, and isoelectric point. The database was constructed from cells kept in resting conditions or stimulated with 59 mM K+ in 2.5 mM Ca2+ or in 0 Ca2+ solution. These manipulations caused statistically significant changes in the degree of phosphorylation of 20 proteins; they were classified as Ca2+-dependent substrates for the phosphorylation or dephosphorylation processes. These changes were also shown in cells stimulated in the presence of the Ca2+ channel activator Bay K 8644. New proteins that show as much as a fivefold increase in their phosphorylation state during cell stimulation have been located with this methodology, as well as many others that had not previously been detected with conventional methods. These experiments provide the first CAIA database of chromaffin cell phosphoproteins; the map constructed with these data will allow the location of specific phosphoproteins and serve as a reference for future ongoing studies. The database will continue to grow to identify more proteins and to facilitate the comparison of complex patterns obtained in different laboratories for normal and transformed pheochromocytoma PC12 cells.     

Spross-Vegetationspunkt und Blattanlage bei Einigen Monokotylen wasserpflanzen (Potamogeton crispus,Heteranthera dubia,Typha angustifolia)

Ohne ZusammenfassungMit 27 Textabbildungen (57 Einzelbildern).D 2.     

On the morphology of spermatozoa of tuco-tucos, Ctenomys (Rodentia: Ctenomyidae): New data and its implications for the evolution of the genus

Sperm morphology was studied in 10 species of the caviomorph rodent Ctenomys. Ctenomys argentinus, C. conoueri, C. dorbigny and C.perrensis had symmetric spermatozoa with paddle-like heads. Ctenomys australis, C. mendocinus, C. porteousi, C. rionegrensis and Ctenomys sp., on the other hand, had spermatozoa with paddle-like heads but with the tail inserted at one side of the central axis and a nuclear caudal extension originating from the base of the head at the opposite side of the insertion of the tail and running parallel to the flagellum; these spermatozoa are referred to as simple-asymmetric. In C. yolandae, a complex-asymmetric morphological type not previously described for the genus was found. This type is characterized by the presence of two nuclear caudal extensions. Symmetric spermatozoa (total length = 52 pm) were shorter than asymmetric (both simple and complex) ones (total length = 87 pm). In spite of these differences, the relative size of heads, midpieces and tails were maintained in the three groups, representing 12%, I I YO and 88% of the average total length, respectively. Within each group of species bearing the same sperm type, a low interspecific variability both in morphological patterns and dimensions of sperm cells was observed. This low interspecific variability associated with the north to south geographical distribution of species having, respectively, symmetric and asymmetric spermatozoa, suggests that these characters appeared at an early stage in evolution of the group, and probably played an important role in the first steps of speciation by promoting reproductive isolation.     

MOLECULAR PHYLOGENETIC ANALYSIS OF LIFE-HISTORY EVOLUTION IN ASTERINID STARFISH

We analyzed phylogenetic relationships among 12 nominal species of starfish in the genera Patiriella and Asterina (Order Valvatida, Family Asterinidae), based on complete sequences for a mitochondrial protein coding gene (cytochrome oxidase subunit I) and five mitochondrial transfer RNA genes (alanine, leucine, asparagine, glutamine, and proline) (1923 bp total). The resulting phylogeny was used to test a series of hypotheses about the evolution of life-history traits. (1) A complex, feeding, planktonic larva is probably ancestral for these starfish, but this is not the most parsimonious reconstruction of ancestral larval states. (2) The feeding larval form was lost at least four times among these species, and three of these losses occurred among members of a single clade. (3) Small adult size evolved before both cases of hermaphroditism and viviparous brooding, but viviparity was not always preceded by an intermediate form of external brooding. (4) An ordered transformation series from feeding planktonic development to viviparous brooding has been predicted for starfish, but we could not find an example of this transformation series. (5) Viviparity evolved recently (< 2 Mya). (6) Both species selection and transformation of lineages may have contributed to the accumulation of species with nonfeeding development among these starfish. (7) Neither Asterina nor Patiriella are monophyletic genera. Larval forms and life-history traits of these starfish have evolved freely under no obvious constraints, contrary to the widely assumed evolutionary conservatism of early development.