Comparative Biochemistry and Physiology Part A: Physiology最新文献

Ten hemoglobins from the teleost Micropogon undulatus were separated in native polyacrylamide gel electrophoresis. In a large sample taken from a tributary of the Chesapeake Bay in Virginia, the distribution of these hemoglobins confirmed a polymorphic condition, which was previously known in North Carolina animals. The intraspecific variation was unrelated to body length, indicating that it is an adult phenomenon. The 10 hemoglobins were expressed in seven different phenotypic banding patterns. Although several morphs were infrequent, no single phenotype comprised a majority of the sample, indicating that the variation is quite great. Several phenotypes differed from those in North Carolina animals and, conversely, several North Carolina phenotypes were not recovered from the Chesapeake Bay sample. This finding suggests that the polymorphism is more complex than observed in either investigation alone. The O₂ binding of hemolysates stripped of co-factors by dialysis revealed functional differences between some, but not all, Chesapeake Bay morphs. The most variable property was O₂ affinity, although a few differences in pH dependence and cooperativity were also observed. The O₂ binding of red blood cell preparations also differed. These differences were not identical to those between dialyzed hemolysates, suggesting that organic PO₄ co-factors may be involved as well.

As part of our ongoing research on cardiac hypoxia tolerance we have conducted ³¹P nuclear magnetic resonance (NMR) studies of isolated, perfused, working hearts from freshwater turtles, animals that are well known for their ability to tolerate prolonged periods of anoxia. A striking feature of turtle heart spectra is an extremely high concentration of NMR visible phosphodiesters (PDEs). Cardiac spectra from mammals, on the other hand, typically exhibit only a small resonance in the PDE region. Our aim in this study was to compare myocardial PDE profiles between the highly hypoxia tolerant western painted turtle (Chrysemys picta bellii) and the relatively hypoxia sensitive softshelled turtle (Trionyx spinifer) in order to begin to test the hypothesis that high constitutive levels of cytosolic PDEs may play a role in conferring hypoxia and ischemia tolerance on the myocardium. We also collected ³¹P-NMR spectra of PCA extracts of tissue from these species and from Kemp's ridley sea turtles (Lepidochelys kempi), as well as spectra from isolated hearts and PCA extracts of redeared sliders (Trachemys [formerly Pseudemys] scripta]). Total NMR visible phosphodiesters make up 24 ± 8.6% of the total NMR visible phosphorus in chrysemys hearts, 20.7 ± 5.9% in Trachemys hearts, but only 12.2 ± 5.1% in Trionyx hearts (P < 0.05). We have identified three distinct PDEs in turtle hearts: glycerophosphorylcholine (GPC); glycerophosphorylethanolamine (GPE); and serine ethanolamine phosphodiester (SEP). SEP is the dominant compound in Chrysemys and Trachemys (79.3 ± 10.2% and 84.7 ± 3.7% of total PDE, respectively), while GPC is most abundant in Trionyx (74.0 ± 4.3% of total PDE) and Lepidochelys (not quantitated). The function of this class of compounds is unclear but it has been suggested that cytosolic PDEs may function as lysophospholipase inhibitors, a role that would decrease the rate of membrane phospholipid turnover. Our comparative data suggest that cytosolic PDEs could play a role in phospholipid sparing during anoxic or ischemic stress in turtles but a direct test of this hypothesis awaits future experimentation.

Hemagglutinins were determined in six species of mosquitoes that are susceptible and refractory to Brugia malayi (Filarioidea: Nematoda). High titers of hemagglutinins were found in the salivary gland extract and in the body fluid of a completely refractory species, Aedes taeniorhynchus, and in partially refractory species, Anopheles quadrimculatus; but low levels of hemagglutinins were also present in the body fluid of Aedes aegypti (Black-eye, Liverpool strain), a susceptible species. Hemagglutinating activity was not found in the other three completely refractory species of mosquitoes, Culex quinquefasciatus, Culex nigripalpus, and Aedes albopictus in which blood coagulated rapidly after ingestion. High titers of hemagglutinins in the salivary glands of Ae. taeniorhynchus and An. quadrimaculatus facilitated rapid movement of sheathed microfilariae from the midgut to the hemocoel. It is suggested that high titers of hemagglutinins present in the hemocoel bound to the glycoconjugates with exposed carbohydrate moieties present on the microfilarial sheaths and developing abnormal larvae (L₁) in the thoracic muscle cells. These hemagglutinin-bound glycoconjugates formed capsules that subsequently stimulated the immune response and resulted in melanization of microfilarial sheaths and sheathed microfilariae in the hemocoel and intracellularly developing abnormal L₁ in the thoracic muscles. Only minimal encapsulation and melanization of B. malayi microfilariae was observed in the hemocoel of the other four species of mosquitoes that lacked hemagglutinins in the salivary glands. The results suggest that tissue specific hemagglutinins are one of several factors of vector susceptibility/refractoriness through immune reactions (encapsulation, activation of prophenoloxidases).

The monophyly of the antarctic fish suborder Notothenioidei and the monophyly of its earliest family the Bovichtidae have been investigated with 12S and 16S mitochondrial DNA sequences. New data from Cottoperca, Pseudaphritis, Harpagifer and several outgroups, in addition to available sequences, show that the bovichtids are paraphyletic. Pseudaphritis is the sister group of all the non-bovichtid notothenioids. The same results are found from two independent genetic markers, the nuclear 28S rDNA and the 12S and 16S mitochondrial rDNA. This reliably refutes a previous hypothesis that placed Pseudaphritis as the sister group of all the remaining notothenioids (including Cottoperca and Bovichtus). Bootstrap analyses show that the Notothenioidei are monophyletic (although members of the suborder Trachinoidei have not been surveyed). Subsequent data from hemoglobin composition confirm the present relationships. After discussions between members of the European Science Foundation (ESF) network during its last two meetings, we point out here some fundamental aspects of comparative biology to improve understanding between the physiologist community and phylogeneticists. The most important points are differences in how the concept of homology is used and differences in the consideration of adaptation. When adaptation is evoked or questioned, endless speculations and untestable scenarios are often developed. We strongly advocate the use of phylogenetic trees for testing hypotheses of adaptation (through multiple character mapping). Such a “research program” in comparative biology has the power to improve knowledge because it can potentially lead to new experiments for testing adaptive hypotheses.

In the early 1960s, the first evidence was reported demonstrating neurally mediated responses in the presence of adrenergic and cholinergic antagonists, leading to the introduction of the concept of non-adrenergic non-cholinergic neurotransmission. The inhibitory component of this part of the autonomic nervous system has been illustrated in numerous organ systems mediating a wide range of physiological events. Since the discovery of these nerves, several substances have been proposed as putative neurotransmitter, with ATP and vasoactive intestinal polypeptide as main candidates. Finally, the ongoing research on the nature of the substance released by these nerves has generated the nitrergic theory proposing nitric oxide as putative neurotransmitter. By now, increasing evidence is reported to support the idea that inhibitory neurons release more neurotransmitters, interacting with each other at pre-and/or postsynaptic levels.

Blood acid-base homeostasis of the hemoglobinless Antarctic teleost Chionodraco hamatus was analyzed by measuring the titratable buffer capacity (β) and carbonic anhydrase (CA) activity in blood smaples. Results were compared with those obtained in the red-blooded temperate fish Anguilla anguilla. Data show that the titratable blood buffer capacity of icefish, in the range of red-blooded teleost species, was significantly higher than that of A. anguilla. Furthermore, plasma inorganic phosphate and reactive sulfhydryl content was significantly higher in the Antarctic species. No enzymatic activity of CA was detected in blood samples of Antarctic fish, suggesting the absence of a blood CA-isozyme.