Directionality of the pressure-difference receiver ears in the northern leopard frog, Rana pipiens pipiens

We studied the directional response of the coupled-eardrum system in the northern leopard frog, Rana pipiens pipiens. Eardrum behavior closely approximates a linear time-invariant system, with a highly correlated input–output relationship between the eardrum pressure difference and the eardrum velocity. Variations in the eardrum transfer function at frequencies below 800 Hz indicate the existence of an extratympanic sound transmission pathway which can interfere with eardrum motions. The eardrum velocity was shown to shift in phase as a function of sound incident angle, which was a direct result of the phase-shift of the eardrum pressure difference. We used two laser-Doppler vibrometers to measure the interaural vibration time difference (IVTD) and the interaural vibration amplitude difference (IVAD) between the motions of the two eardrums. The coupled-eardrum system enhanced the IVTD and IVAD by a factor of 3 and 3 dB, respectively, when compared to an isolated-eardrum system of the same size. Our findings are consistent with the time-delay sensitivity of other coupled-eardrum systems such as those found in crickets and flies.     

Surface properties of Culex pipiens pipiens eggs and the behaviour of the female during egg-raft assembly

ABSTRACT. Eggs of the mosquito Culex pipiens pipiens L. float vertically because a flexible corolla at the anterior (lower) pole has a hydrophilic lower surface and a hydrofuge upper surface. The corolla assumes different configurations in the egg raft; the surface forces operating on eggs are briefly analysed. The outer surface of the chorion is hydrofuge, but in eggs floating on water there is a jacket of water surrounding the egg in the spaces between the tubercles of the exochorion, below the meshwork connecting them. The humidity over natural water can be low enough to cause continuous evaporation from the jacket; water is replenished through openings within the lower surface of the corolla. The disposition of the corolla prevents the uptake of materials confined to the water surface outside, but the mechanism could concentrate solutes in the jacket, with possible deleterious consequences. Eggs are joined together by inter-digitation of the regularly spaced tubercles. The behaviour by which the female assembles the egg raft is described in detail, explaining the form of the raft and the patterns in which eggs are packed.     

Morphology of neurons in the torus semicircularis of the northern leopard frog,Rana pipiens pipiens

The neuronal morphology of the torus semicircularis of the northern leopard frog, Rana pipiens pipiens, was examined in Golgi-impregnated material. Neurons in each of the five subdivisions of the torus semicircularis (Potter, '65a) have distinct morphologies which are characteristic of the subdivision. Laminar nucleus neurons are mostly multipolar with spherical or ovoidal somata and smooth dendrites oriented primarily parallel and perpendicular to the cell laminae. Principal nucleus neurons have variable soma shapes with short dendrites ( < 100 μm) radiating in all directions. In the magnocellular nucleus, there are three major cell types: neurons characterized by small, spherical-shaped somata, with short, thin, radiating dendrites and many varicosities; bi- or tripolar neurons with ovoidal somata, and long (100–200 μm) and smooth dendrites orienting primarily dorsoventrally and mediolaterally; and multipolar neurons with triangular-shaped somata and very long (200–350 μm) dendrites, which are either smooth or highly spiny. Neurons in the commissural nucleus are mostly multipolar cells with ovoidal somata and beaded dendrites projecting mostly dorsally and ventrally. The subependymal midline nucleus contains mostly uni- or bipolar neurons with small ovoidal somata and straight, spiny dendrites. In addition to revealing the morphological features of neurons in the torus, the counterstained material shows further cytoarchitectural organization of the principal nucleus, i.e., the presence of a circular lamellar organization. The functional significance of these anatomical features is discussed.     

Directionality of phase locking in auditory nerve fibers of the leopard frog Rana pipiens pipiens

Summary A dorsal approach to the eighth nerve and free-field stimulation were used to investigate the effect of sound direction and intensity on phase locking in auditory nerve fibers of the leopard frog Rana pipiens pipiens.Tuning curves of 75 auditory neurons were analyzed (Fig. 2). Amphibian papillar neurons, but not basilar papillar neurons, exhibit significant phase locking to short tone bursts at the characteristic frequency (CF), the degree of phase locking (vector strength) decreasing with the neuron's CF (Figs. 3, 4 and 10E). Vector strength increases with sound pressure level to saturate about 20 dB above threshold, while the preferred firing phase is only slightly affected (Figs. 5 and 6).In contrast, sound direction hardly affects vector strength (Figs. 7, 8, 9A and 10A and C), but has a strong influence on the preferred firing phase (Figs. 7, 8, 9B and C, 10B and D): With respect to anterior tone presentation there are phase lags for ipsilateral and phase leads for posterior and contralateral presentation. Phase differences between both ears show a sinusoidal or cardioid/ovoidal directional characteristic; maximum differences are found with antero-lateral tone presentation (Fig. 11). The directionality of phase locking decreases with the neuron's CF (Fig. 10F) and only slightly changes with sound pressure level (Fig. 12). Thus, phase locking of amphibian papilla neurons can potentially provide intensity-independent information for sound localization.Abbreviations SPL sound pressure level - FTC frequency threshold curve - CF characteristic frequency - TF test frequency - VS vector strength - AP amphibian papilla - BP basilar papilla     

Viability of Culex pipiens pipiens eggs affected by nutrition and aposymbiosis

Raisins were a better source of carbohydrate than sucrose for reproduction by autogenous Culex pipiens. A blood meal increased the number of eggs per raft from 49 autogenously to 114. Eggs of aposymbiotic females produced autogenously did not hatch, but 34% of the eggs were viable if the mosquitoes fed on chickens. With repeated blood meals the number of eggs per raft and the rate of embryonation and hatching declined in each successive gonotrophic cycle. In about $\text{1}\text{4}$ of the unhatched eggs of normal females there were no fully developed embryos, while many more of the unhatched eggs of aposymbiotic females contained no evidence of embryonic development. After the fifth blood meal, neither normal nor aposymbiotic insects oviposited. The ovaries of the nulliparous females contained approximately 10% of the potential number of mature terminal oocytes. Proximally in the ovarioles there were dilatations and coiled tracheoles indicating egg resorption. There were fewer parous follicles in aposymbiotic than in normal females. Larval rearing water, i.e., infusion in which larvae had been reared, was more attractive than fresh infusion for oviposition by normal, autogenous mosquitoes. The degree of embryonation of the eggs was lower and the hatching success rate was poorer in fresh infusion than in larval rearing water.     

Speciation in the Rana pipiens Complex

The long held view that leopard frogs (Rana pipiens complex)are a single widely distributed species is not correct. Fivesibling species are currently recognizable. These findings haveimportant implications on the use of leopard frogs in experimentalresearch.     

Cytodifferentiation in the Rana pipiens oocyte

Summary In early diplotene frog oocytes incubated to illustrate thiamine pyrophosphatase (TPPase) activity, reaction product is uniformly distributed within the compartments of the endoplasmic reticulum and nuclear envelope as well as within the saccules and small vesicles comprising the dictyosomes. With continued oocyte development the reaction product becomes concentrated in localized regions of the dictyosome saccules. Eventually, the enzyme is no longer apparent within the endoplasmic reticulum, but is concentrated in the cisternae of the inner dictyosome saccules. The variations noted suggest that the enzyme is synthesized early in diplotene by the endoplasmic reticulum and is subsequently transported to the Golgi apparatus where it is consistently observed at later developmental stages. TPPase activity is also present in the Golgi apparatus of follicle and theca cells as well as in ovarian epithelial cells. The enzyme is also detected in micropinocytotic vesicles contained within the cells comprising the follicle envelope and in intercellular spaces of the follicle. Horseradish peroxidase injected into the coelomic cavity is transported via micropinocytotic vesicles into and through the cells comprising the follicle envelope and in intercellular spaces. The exogenous protein is not found even after a prolonged time period in early diplotene oocytes. The protein is, however, present in large spherical and tubular vesicles in the cortex of vitellogenic oocytes approximately 500 microns in diameter. The possible functional role of the enzyme TPPase during oogenesis is discussed.This investigation was supported by a research grant from the National Science Foundation (GB-8736).     

Effect of photoperiod and temperature on ovarian development in Culex pipiens pipiens

Culex pipiens pipiens is one member of the C. pipiens complex which diapauses as an adult. Experiments show that photoperiod and temperature both affect the development of the ovarian follicles. Females reared at a 15 hr photophase and 22°C show an increase in follicle size over the first 8 days after emergence. Females reared at 10 hr and 22°C do not show an increase in follicle size. Follicle size does not gradually increase as photophase is lengthened. Rather, at photophases between 9 and 12 hr the follicles remain small and at photophases of 13 hr or more, the increase is seen. An increase in temperature tends to increase follicle size regardless of the photophase. However, a strong interaction between photophase and temperature is evident.No evidence exists for a single ‘critical stage’. Instead, the final follicle size is dependent, in a complex manner, upon the photophase experienced by all the stages. The final size is not a result of a cumulative effect.Photoperiod, independent of temperature, shows a marked effect on blood feeding. Females reared at 15 hr feed much more readily than those reared at 10 hr. Those 10 hr females that feed show no signs of gonotrophic dissociation.     

Ultrastructure of the jugular body of Rana pipiens

Summary The jugular bodies in adult Rana pipiens, are surrounded by a capsule of mesothelium and connective tissue, and their parenchyma consists of cell cords arranged in a sinusoidal network. The cell cords are formed by irregular reticular cells, showing numerous filaments and joined together by zonulae adhaerents. The intercellular spaces are filled by reticular fibres and free cells. These latter are small and medium lymphocytes, lymphoblasts, and developing and mature plasma cells. Additionally, free macrophages, neutrophils and acidophils also occur. Sinusoidal blood vessels show thin walls with numerous filaments and pinocytotic vesicles. They exhibit a discontinuous basement membrane, and tight junctions frequently occur between endothelial cells. Occasionally, lymphatic vessels are found and the innervation is principally vasomotor, although nerve endings appear remarkably near reticular cells and lymphocytes. The jugular bodies of adult R. pipiens are plasma cell and antibody-forming organs, whose functional significance is discussed in relation to their ultrastructural organization.     

Esterases in the mosquito Culex pipiens pipiens L.: Formal genetics and polymorphism of adult esterases

The genetics of two esterase loci active in autogenous adults of the mosquito Culex pipiens pipiens L. has been studied by means of starch gel electrophoresis. Three alleles at the Est-1 locus and eight at the Est-2 locus are described. Both loci have a null allele. Active alleles are codominant and there is no hybrid enzyme in heterozygotes. The Est-1 locus codes esterases preferentially hydrolyzing -naphthylacetate and the Est-2 locus esterases preferentially hydrolyzing -naphthylacetate. Strains homozygous for both loci were selected. Linkage studies of the two loci have shown that they are not sex linked but are linked to each other, the crossover frequency being 8.6%. The polymorphism of two laboratory and two natural populations is described for both loci. Phenotypic distributions are in good agreement with Hardy-Weinberg expectations.This work was conducted at the Université des Sciences et Techniques du Languedoc (Laboratoire de Génétique Expérimentale des Populations), Montpellier, France, in partial fulfillment of the requirements for the degree of Docteur de spécialité.     

Differentiation of the Opercular Integument in Rana pipiens

The opercular skin develops in its own specific fashion differing from other areas of the integument previously studied. It remains typically larval until it is sloughed off.
The opercular integument first deviates from the common skin development pathway at stage XV [1] when the rough endoplasmic reticulum. proliferates to form a large part of the cytoplasm and is accompanied by numerous large mitochondria. The Golgi apparatus becomes very large and many vesicles are found in the cytoplasm.
The mitochondria become large and swollen with atypical structure. The cytoplasm becomes highly vesiculated. Degeneration begins at the dermal edge of the basement lamella and proceeds toward the epidermis, being marked by the disorientation of collagen fibrils.
Epidermal cells become progressively more necrotic having cytoplasm consisting mainly of residual bodies and vesicles: the nucleus is the last cellular structure to undergo autolysis.
The dermal cells are the first to be lost (stages XVIII–XIX). The opercular integument is shed as a sheet of cells at stage XX when the forelimbs emerge. The necrotic zone of the opercular integument appears to stop at the junction of the body skin and the skin of the forelimb.     

Cultivation of the mosquito Culex pipiens pipiens f. molestus (Diptera, Culicidae) without blood feeding

Optimal photoperiod and temperature conditions for rearing of the mosquito Culex pipiens pipiens f. molestus without blood feeding (through autogenous eggs) were experimentally substantiated. This method may be used in order to obtain relatively small quantities of mosquitoes or for preservation of a laboratory colony. Optimal conditions of rearing are the short-day photoperiod (12 h of light per day) at 20 degrees C, when 87 % of the females produce autogenous eggs rafts consisting of 52 eggs on the average. It should be emphasized that larval food must be adequate and abundant, because the quality and quantity of it has a pronounced effect on the autogeny rate.     

Esterase polymorphism and sensitivity to dursban organophosphorus insecticide in Culex pipiens pipiens populations

Esterase polymorphism and Dursban (O,O-dimethyl-2-pyridylphosphorothioate) sensitivity have been investigated in 12 natural populations and three laboratory strains of Culex pipiens pipiens. This mosquito has two esterase loci, Est-1 and Est-2, which were shown to code esterases of the B group (aliesterases) but not cholinesterases. No correlation between Est-1 polymorphism and Dursban sensitivity was found, but the increase of the Est-2(0.64) allele in the populations less sensitive to Dursban was highly significant (r = -0.9850 for 6 df).