Contractile behavior of the forelimb digital flexors during steady-state locomotion in horses (Equus caballus): an initial test of muscle architectural hypotheses about in vivo function

The forelimb digital flexors of the horse display remarkable diversity in muscle architecture despite each muscle-tendon unit having a similar mechanical advantage across the fetlock joint. We focus on two distinct muscles of the digital flexor system: short compartment deep digital flexor (DDF(sc)) and the superficial digital flexor (SDF). The objectives were to investigate force-length behavior and work performance of these two muscles in vivo during locomotion, and to determine how muscle architecture contributes to in vivo function in this system. We directly recorded muscle force (via tendon strain gauges) and muscle fascicle length (via sonomicrometry crystals) as horses walked (1.7 m s(-1)), trotted (4.1 m s(-1)) and cantered (7.0 m s(-1)) on a motorized treadmill. Over the range of gaits and speeds, DDF(sc) fascicles shortened while producing relatively low force, generating modest positive net work. In contrast, SDF fascicles initially shortened, then lengthened while producing high force, resulting in substantial negative net work. These findings suggest the long fibered, unipennate DDF(sc) supplements mechanical work during running, whereas the short fibered, multipennate SDF is specialized for economical high force and enhanced elastic energy storage. Apparent in vivo functions match well with the distinct architectural features of each muscle.     

Short dry spells in the wet season increase mortality of tropical pioneer seedlings

Variation in plant species performance in response to water availability offers a potential axis for temporal and spatial habitat partitioning and may therefore affect community composition in tropical forests. We hypothesized that short dry spells during the wet season are a significant source of mortality for the newly emerging seedlings of pioneer species that recruit in treefall gaps in tropical forests. An analysis of a 49-year rainfall record for three forests across a rainfall gradient in central Panama confirmed that dry spells of ≥10 days during the wet season occur on average once a year in a deciduous forest, and once every other year in a semi-deciduous moist and an evergreen wet forest. The effect of wet season dry spells on the recruitment of pioneers was investigated by comparing seedling survival in rain-protected dry plots and irrigated control plots in four large artificially created treefall gaps in a semi-deciduous tropical forest. In rain-protected plots surface soil layers dried rapidly, leading to a strong gradient in water potential within the upper 10 cm of soil. Seedling survival for six pioneer species was significantly lower in rain-protected than in irrigated control plots after only 4 days. The strength of the irrigation effect differed among species, and first became apparent 3–10 days after treatments started. Root allocation patterns were significantly, or marginally significantly, different between species and between two groups of larger and smaller seeded species. However, they were not correlated with seedling drought sensitivity, suggesting allocation is not a key trait for drought sensitivity in pioneer seedlings. Our data provide strong evidence that short dry spells in the wet season differentially affect seedling survivorship of pioneer species, and may therefore have important implications to seedling demography and community dynamics.     

What happens when stems are embolized in a centrifuge? Testing the cavitron theory

Vulnerability curves (VCs) measure the ability of vessels to retain metastable water without embolisms that lower the hydraulic conductivity of stems. The fastest method of measuring VCs is the centrifuge technique and the Cochard cavitron is a method that allows measurement of hydraulic conductivity of stems while they are spinning. This paper describes the pattern of embolism that results after spinning the stems of hybrid aspen (Populus tremula×P. tremuloides) and two hybrid cottonwoods (P38P38 P. balsamifera×P. simonii and Northwest, which is a hybrid of P. deltoides×P. balsamifera). It is recognized that the pattern of embolism induced in a centrifuge ought to differ from the pattern during natural dehydration of plants because the profiles of tension vs distance greatly differ under the two modes of inducing stress. The pattern of embolism was visualized by a staining technique and quantified by traditional measurements of percentage loss conductivity (PLC) performed on subsample segments excised from spun stems. We found a pattern of embolism approximating that expected from theory: (1) PLC near the axis of rotation exceeded the average; (2) PLC was quite high near the ends of the stems, even though tension ought to be zero; (3) large vessels cavitated before small vessels; (4) more embolism occurred near the base than near the apex of the stems. However, we could not always scale up from subsample conductivity and PLC to whole‐stem conductivity. This pattern of embolism is interpreted in terms of vessel diameter and vessel length.     

SEASONAL OCCURRENCE OF XYLEM EMBOLISM IN SUGAR MAPLE (ACER SACCHARUM)

Xylem embolism, the reduction of water flow by air-filled vessels, was measured in a stand of 5- to 8-year-old sugar maple (Acer saccharum Marsh.) saplings growing in a nursery bed in northwestern Vermont. Embolism was quantified as percentage loss in hydraulic conductivity of trunk and branch segments relative to maximum values obtained by removing air from vessels by repeated high pressure (173 kPa) perfusions. Ten segments per tree were cut from 6 trees for each of 11 measurement periods spaced at roughly monthly intervals from May 1986 to June 1987. During the 1986 growing season, embolism increased significantly from 11 to 31% in the larger branches and trunk (segment diameter #8805;0.5 cm), but remained at about 10% in twigs (segment diameter <0.5 cm). This was unexpected because the greatest water stess and thus potential for embolism occurs in twigs. During the winter, embolism increased throughout the trees and the trend with diameter was reversed; by February, small twigs were 84% embolized vs. 69% for larger branches and trunk. Dye perfusions showed that winter embolism in trunks was localized on the south side; this may have resulted from water loss by sublimation or evaporation in the absence of water uptake. Beginning in late March, embolism decreased throughout the trees to approximately 20% in June. This decrease was associated with positive xylem pressure of at least 16 kPa which may have originated in the roots, because weather conditions at the time were unfavorable for the generation of stem pressures characteristic of Acer species in early spring.     

Interactions of divalent cations with single calcium channels from rat brain synaptosomes

Voltage-dependent calcium channels from a rat brain membrane preparation ("synaptosomes") were incorporated into planar lipid bilayers. The effects of calcium, barium, strontium, manganese, and cadmium ions on the amplitudes and kinetics of single channel currents were examined. The order of single channel conductances was gBa greater than gSr greater than gMn, which was the inverse of the order of the mean channel open times: TMn greater than TCa = TSr greater than TBa. In contrast, the identity of the charge carrier had little or no effect on the mean closed times of the channel. Manganese, in the absence of other permeant ions, can pass through single channels (gMn = 4 pS). However, when added to a solution that contained another type of permeant divalent cation, manganese reduced the single channel current in a voltage-dependent manner. Cadmium, a potent blocker of macroscopic "ensemble" calcium currents in many preparations, reduced the current through an open channel in a manner consistent with Cd ions both not being measurably permeant and interacting with a single site. The permeant ions competed with cadmium for this site with the following order: Mn greater than Sr = Ca greater than Ba. These results are consistent with the existence of no less than one divalent cation binding site in the channel that regulates ion permeation.     

Altered synthesis of heparan sulfate proteoglycans at low sulfate concentration

The structural alterations in heparan sulfate produced by sulfate deprivation were studied in cell cultures of the Engelbreth-Holm-Swarm tumor. Tumor cells were labeled in vitro with [3H]glucosamine and/or [35S]sulfate in media containing either 300 microM MgSO4 or no added carrier sulfate, and the newly synthesized proteoglycans isolated by chromatography on DEAE-Sephacel. The proteoglycans isolated from low sulfate cultures showed a reduced affinity for the column eluting at lower salt concentrations compared with the proteoglycans isolated from cultures containing sulfate, suggesting that the former were undersulfated. Analysis of the isolated heparan sulfate side chains indicated that two pools of heparan sulfate were present which differed in their degree of sulfation. Both pools were synthesized by both high sulfate and low sulfate cultures, but the highly sulfated pool was the predominant form produced in sulfate containing cultures, while the undersulfated pool was the predominant form synthesized in low sulfate cultures. The more sulfated pool contained more N-sulfate than the less sulfated pool. Few if any free amino groups were detected in either pool, suggesting that the initial deacetylation step in the biosynthesis of heparan sulfate is tightly coupled to the N-sulfation step in the cells.     

Pandinus imperator scorpion venom blocks voltage-gated potassium channels in GH3 cells

We examined the effects of Pandinus imperator scorpion venom on voltage-gated potassium channels in cultured clonal rat anterior pituitary cells (GH3 cells) using the gigohm-seal voltage-clamp method in the whole-cell configuration. We found that Pandinus venom blocks the voltage-gated potassium channels of GH3 cells in a voltage-dependent and dose-dependent manner. Crude venom in concentrations of 50-500 micrograms/ml produced 50-70% block of potassium currents measured at -20 mV, compared with 25-60% block measured at +50 mV. The venom both decreased the peak potassium current and shifted the voltage dependence of potassium current activation to more positive potentials. Pandinus venom affected potassium channel kinetics by slowing channel opening, speeding deactivation slightly, and increasing inactivation rates. Potassium currents in cells exposed to Pandinus venom did not recover control amplitudes or kinetics even after 20-40 min of washing with venom-free solution. The concentration dependence of crude venom block indicates that the toxins it contains are effective in the nanomolar range of concentrations. The effects of Pandinus venom were mimicked by zinc at concentrations less than or equal to 0.2 mM. Block of potassium current by zinc was voltage dependent and resembled Pandinus venom block, except that block by zinc was rapidly reversible. Since zinc is found in crude Pandinus venom, it could be important in the interaction of the venom with the potassium channel. We conclude that Pandinus venom contains toxins that bind tightly to voltage-dependent potassium channels in GH3 cells. Because of its high affinity for voltage-gated potassium channels and its irreversibility, Pandinus venom may be useful in the isolation, mapping, and characterization of voltage-gated potassium channels.     

Vulnerability of xylem to embolism in a mangrove vs an inland species of Rhizophoraceae

Vulnerability of xylem conduits to cavitation and embolism was compared in two species of Rhizophoraceae, the mangrove Rhizophora mangle L. and the tropical moist-forest Cassipourea elliptica (Sw.) Poir. Cavitation (water column breakage preceeding embolism) was monitored by ultrasonic detection; embolism was quantified by its reduction of xylem hydraulic conductivity. Acoustic data were not predictive of loss in hydraulic conductivity, probably because signals from cavitating vessels were swamped by more numerous ones from cavitating fibers. Rhizophora mangle was the less vulnerable to embolism of the two species, losing 80% of its hydraulic conductivity between – 6.0 and – 7.0 MPa. Cassipourea elliptica lost conductivity in linear proportion to decreasing xylem pressure from – 0.5 to – 7.0 MPa. Species vulnerability correlated closely with physiological demands of habitat; the mangrove Rhizophora mangle had field xylem pressures between – 2.5 and – 4.0 MPa. whereas the minimum for Cassipourea elliptica was – 1.6 MPa. Differences in vulnerability between species could be accounted for by differences in the measured air permeability of intervessel pit membranes. According to this explanation, embolism occurs when air enters a water-filled vessel from a neighboring air-filled one via pores in shared pit membranes.     

Cerebellar neurones: Differentiation and modulation of sensitivity to excitotoxic treatment

The neurite outgrowth and adhesion complex (NOAC), isolated from rabbit sera has been dissociated in its major components by reverse-phase chromatography in HPLC by using a C₁₈ column. SDS-PAGE analisys of the active fractions revealed the presence of three major bands of approximately 100, 70 and 50 kDa. Studies on the biological activity of NOAC were carried out on rat cerebellar granule cells. NOAC-cultured cells exhibit a marked resistance to excitotoxic stimuli carried by glutamate.