The nervous system of Tricladida. I. Neuroanatomy ofProcerodes littoralis (Maricola,Procerodidae): An immunocytochemical study

The organization of the nervous system ofProcerodes littoralis (Tricladida, Maricola, Procerodidae) was studied by immunocytochemistry, using antibodies to authentic flatworm neuropeptide F (NPF) (Moniezia expansa). Compared to earlier investigations of the neuroanatomy of tricladid flatworms, the pattern of NPF immunoreactivity inProcerodes littoralis reveals differences in the following respects: 1. Shape and structure of the brain. 2. Number and composition of longitudinal nerve cords. 3. Shape of branches of, and transverse connections between, main ventral nerve cords. 4. Composition of the pharyngeal nervous system. The rich innervation by NPF immunoreactive (IR) fibres and cells of the subepithelial muscle layer, the pharynx musculature and the musculature of the male copulatory apparatus indicates a neurotransmitter or neuromodulatory influence on muscular activity.     

Lectin-induced deciduoma formation in the pseudopregnant rat

Decidual cell induction in the pseudopregnant rat was examined in this study using the lectin concanavalin A (ConA). The histochemical binding of the lectin to the uterine cell surface at the time of deciduomatic induction was also studied. ConA was found to induce significant deciduomata (decidual-like tissue) in the uterine horn when injected intraluminally on day 5 of pseudopregnancy (PSP). ConA-induced deciduomata appeared as a series of discrete nodules in the uterine horn, reminiscent of the anatomical appearance of normal embryo implantation sites. Deciduoma induction by ConA was greatly reduced by pre-absorption of the lectin with its competitive sugar. Lectin histochemistry revealed binding of ConA to the cell surface on day 5 of PSP. Pre-absorption of the lectin with its competitive sugar also significantly reduced surface binding of the lectin, and this finding may be correlated with the greatly reduced ability of the pre-absorbed lectin to induce deciduomata. Possible mechanisms for the induction of deciduomata by lectins are considered.     

QUANTITATIVE GENETICS OF SEQUENTIAL LIFE-HISTORY AND JUVENILE TRAITS IN THE PARTIALLY SELFING PERENNIAL,AQUILEGIA CAERULEA

We determined the genetic basis of several traits related to overall fitness of Aquilegia caerulea, a perennial herb of the Rocky Mountains in western North America. To obtain measures of heritability relevant to the evolutionary potential of wild populations, we performed full and partial diallel crosses and studied progeny performance in the field. Based on a joint analysis of two designs with a total of 18 parents and 102 crosses, we detected significant maternal variance for seed mass and emergence time, but this component was negligible for later-expressed traits. Low heritability and evidence that maternal effects on seed mass are largely environmental suggest that in this population there is little evolutionary potential for change in seed mass under conditions experienced during the study. Seed mass varied depending on particular combinations of parents and cross direction. Such an interaction can have several different biological interpretations, including that particular maternal parents selectively provision embryos sired by particular pollen genotypes. Width of the first true leaf after 4 wk of growth and leaf size of juvenile plants at years one and two were significantly heritable and positively genetically correlated. Juvenile survival exhibited significant dominance variance, as expected from evidence of inbreeding depression in this trait. In contrast, for other traits that exhibit inbreeding depression in this population (seed mass and third-year leaf size), dominance variance was negligible.     

Isolation and characterization of restriction endonuclease EclHKI from an Enterobacter cloacae strain

An Enterobacter cloacae strain, producing restriction enzyme EclHKI, was isolated from a decaying potato. The enzyme is an isoschizomer of Eam1105I, which recognizes and cleaves GACNNN!NNGTC. EclHKI was produced at high activity (4×10⁴ U/g wet cells) and was purified from contaminants which interfere with restriction digestion by passing the cell lysate through DEAE-Sephacel and heparin columns. Activity was optimal at 37°C in a medium salt buffer.H.-Y. Chan, Y.-C. Chan and P.-C. Shaw are with the Department of Biochemistry, The Chinese University of Hong Kong, Hong Kong: K.-M. Kam is with the Institute of Pathology, Sai Ying Pun Jockey Club Clinic, Hong Kong.     

A Neurofilament-Associated Kinase Phosphorylates Only a Subset of Sites in the Tail of Chicken Midsize Neurofilament Protein

Although neurofilaments are among the most highly phosphorylated proteins extant, relatively little is known about the kinases involved in their phosphorylation. The majority of the phosphates present on the two higher-molecular-mass neurofilament subunits are added to multiply repeated sequence motifs in the tail. We have examined the specificity of a neurofilament-associated kinase (NFAK) partially purified from chicken spinal cord that selectively phosphorylates the middle-molecular-mass neurofilament subunit, NF-M. Two-dimensional phosphopeptide mapping of ³²P-labeled NF-M shows that, in vitro, NFAK phosphorylates a subset of peptides phosphorylated in vivo in cultured neurons. The absence of a complete complement of labeled phosphopeptides following in vitro phosphorylation, compared with phosphorylation in vivo, is not due to a lack of availability of phosphorylation sites because the same maps are obtained when enzymatically dephosphorylated NF-M is used as an in vitro substrate. Phosphopeptide maps from in vitro-phosphorylated NF-M and those from a recombinant fusion protein containing only a segment of the tail piece of chicken NF-M reveal identical labeled peptides. The fusion protein lacks a segment containing 17 KXX(S/T)P putative phosphorylation sites contained in the tail of chicken NF-M but contains a segment that includes four KSPs and a KSD site also present in the intact tail. These results suggest (a) that NFAK mediates the phosphorylation of some, but not all, potential phosphorylation sites within the tail of NF-M and (b) that multiple kinases are necessary for complete phosphorylation of the NF-M tail.     

QUANTITATIVE GENETICS OF RESPONSE TO COMPETITORS IN NEMOPHILA MENZIESII: A GREENHOUSE STUDY

To determine the potential for adaptation to a local biotic environment, we examined the magnitude and nature of genetic variation in response to neighboring plants within a natural population of the native California annual, Nemophila menziesii. A total of 22 plants from a natural population were crossed in three reciprocal factorials. The progeny were grown in a greenhouse in nine treatments that varied in conspecific density and in the density of a naturally co-occurring grass species, Bromus diandrus. Increasing the density of each species significantly reduced individual survival, fruit number, and dry weight. Among survivors, we found small to moderate heritability of dry weight within treatments. Additive genetic correlations (r_A) of dry weight between competitive regimes were generally large and positive. In no case were they significantly different from 1, as expected under the null hypothesis that the relative performance of the genotypes under consideration is the same in all environments. On the basis of these results, we cannot conclude that the structure of genetic covariation within this population would promote genetic differentiation in response to locally varying conditions of density of these two species. Aspects of the experiment that may have compromised our ability to detect r_A differing from 1 are discussed.     

The inheritance of vegetative growth traits in strawberries (Fragaria x ananassa) grown at low temperatures and their relationship to field productivity

The genetic relationship between vegetative growth at low temperatures and productivity was investigated for strawberries grown in controlled and field environments. Genotypes from 20 biparental crosses were grown in controlled environments with 11°, 14°, and 17 °C days, 11 °C nights, and 11-h daylength to simulate a range of winter growing conditions expected in mediterranean environments. Individual plants were scored for two initial runner traits and eight vegetative growth traits. Significant main effects of temperature and cross were detected for all growth chamber traits, and conservative estimates of the broad sense heritability (h²) for these traits were 0.10–0.28. None of the temperature x cross interaction effects were significant, suggesting that genetic potential for vegetative growth and vigor is expressed similarly at low and optimal growing temperatures. Highly significant genetic correlations were detected between many growth chamber trait pairs, indicating pleiotropic effects for the genes that condition these traits. Complementary field trials were established, and individual plants were scored for traits that describe yield, production pattern, and plant size. Significant negative genetic correlations were detected between traits that describe growth in the chambers and early production in the field trials, but genetic correlations between chamber growth traits and mid-season or total production were significantly positive and occasionally large. Several of the yield and field growth variables were genetically correlated to initial runner plant traits, suggesting that indirect selection using traits scored in the nursery can be used to improve yield and modify production pattern in the field.     

Palmitylation of an amino-terminal cysteine motif of protein tyrosine kinases p56lck and p59fyn mediates interaction with glycosyl-phosphatidylinositol-anchored proteins.

Cross-linking of glycosyl-phosphatidylinositol (GPI)-anchored membrane proteins on T cells can trigger cell activation. We and others have shown an association between GPI-anchored proteins and the protein tyrosine kinases (PTKs) p56lck and p59fyn, suggesting a pathway for signaling through GPI-anchored proteins. Studies of decay-accelerating factor (DAF) or CD59 in either the C32 cell line or the HeLa cell line transfected with PTK cDNA demonstrated that the GPI-anchored proteins associated noncovalently with p56lck and p59fyn but not with p60src. Nonmyristylated versions of p56lck and p59fyn also failed to associate with the GPI-anchored proteins. Mutational analysis of the PTK demonstrated that the association with the GPI-anchored proteins mapped to the unique amino-terminal domains of the PTK. A chimeric PTK consisting of the 10 amino-terminal residues of p56lck or p59fyn replacing the corresponding amino acids in p60src was sufficient for association with DAF, but the converse constructs containing the first 10 amino acids of p60src plus the remainder of p56lck or p59fyn did not associate with DAF. Mutation of cysteine to serine at positions 3 and 6 in p59fyn or positions 3 and 5 in p56lck abolished the association of these kinases with DAF. Mutation of serine to cysteine at positions 3 and 6 in p60src conferred on p60src the ability to associate with DAF. Direct labeling with [3H]palmitate demonstrated palmitylation of this amino-terminal cysteine motif in p56lck. Thus, palmitylation of the amino-terminal cysteine residue(s) together with myristylation of the amino-terminal glycine residue defines important motifs for the association of PTKs with GPI-anchored proteins.     

THE COMPARISON OF QUANTITATIVE GENETIC PARAMETERS BETWEEN POPULATIONS

A statistical method for comparing matrices of genetic variation and covariation between groups (e.g., species, populations, a single population grown in distinct environments) is proposed. This maximum-likelihood method provides a test of the overall null hypothesis that two covariance component matrices are identical. Moreover, when the overall null hypothesis is rejected, the method provides a framework for isolating the particular components that differ significantly between the groups. Simulation studies reveal that discouragingly large experiments are necessary to obtain acceptable power for comparing genetic covariance component matrices. For example, even in cases of a single trait measured on 900 individuals in a nested design of 100 sires and three dams per sire in each population, the power was only about 0.5 when additive genetic variance differed by a factor of 2.5. Nevertheless, this flexible method makes valid comparison of covariance component matrices possible.