Getting to the end of RNA: Structural analysis of protein recognition of 5′ and 3′ termini

The specific recognition by proteins of the 5′ and 3′ ends of RNA molecules is an important facet of many cellular processes, including RNA maturation, regulation of translation initiation and control of gene expression by degradation and RNA interference. The aim of this review is to survey recent structural analyses of protein binding domains that specifically bind to the extreme 5′ or 3′ termini of RNA. For reasons of space and because their interactions are also governed by catalytic considerations, we have excluded enzymes that modify the 5′ and 3′ extremities of RNA. It is clear that there is enormous structural diversity among the proteins that have evolved to bind to the ends of RNA molecules. Moreover, they commonly exhibit conformational flexibility that appears to be important for binding and regulation of the interaction. This flexibility has sometimes complicated the interpretation of structural results and presents significant challenges for future investigations.     

Microperfusion Technique to Investigate Regulation of Microvessel Permeability in Rat Mesentery

Experiments to measure the permeability properties of individually perfused microvessels provide a bridge between investigation of molecular and cellular mechanisms regulating vascular permeability in cultured endothelial cell monolayers and the functional exchange properties of whole microvascular beds. A method to cannulate and perfuse venular microvessels of rat mesentery and measure the hydraulic conductivity of the microvessel wall is described. The main equipment needed includes an intravital microscope with a large modified stage that supports micromanipulators to position three different microtools: (1) a beveled glass micropipette to cannulate and perfuse the microvessel; (2) a glass micro-occluder to transiently block perfusion and enable measurement of transvascular water flow movement at a measured hydrostatic pressure, and (3) a blunt glass rod to stabilize the mesenteric tissue at the site of cannulation. The modified Landis micro-occlusion technique uses red cells suspended in the artificial perfusate as markers of transvascular fluid movement, and also enables repeated measurements of these flows as experimental conditions are changed and hydrostatic and colloid osmotic pressure difference across the microvessels are carefully controlled. Measurements of hydraulic conductivity first using a control perfusate, then after re-cannulation of the same microvessel with the test perfusates enable paired comparisons of the microvessel response under these well-controlled conditions. Attempts to extend the method to microvessels in the mesentery of mice with genetic modifications expected to modify vascular permeability were severely limited because of the absence of long straight and unbranched microvessels in the mouse mesentery, but the recent availability of the rats with similar genetic modifications using the CRISPR/Cas9 technology is expected to open new areas of investigation where the methods described herein can be applied.     

Herbivory in a spider through exploitation of an ant–plant mutualism

Spiders are thought to be strict predators [1]. We describe a novel exception: Bagheera kiplingi, a Neotropical jumping spider (Salticidae) that exploits a well-studied ant–plant mutualism, is predominantly herbivorous. From behavioral field observations and stable-isotope analyses, we show that the main diet of this host-specific spider comprises specialized leaf tips (Beltian food bodies; Figure 1A) from Vachellia spp. ant-acacias (formerly Acacia spp.), structures traded for protection in the plant's coevolved mutualism with Pseudomyrmex spp. ants that inhabit its hollow thorns [2]. This is the first report of a spider that feeds primarily and deliberately on plants.     

Tonic Sympathetic Nervous System Inhibition of Insulin Secretion Is Diminished in Obese Zucker Rats

It has long been known that the central nervous system (CNS) directly affects pancreatic insulin release. This study was undertaken to determine the effect of the CNS on pancreatic insulin release in three-month-old female lean (Fa/Fa) and hyperinsulinemic obese (fa/fa) Zucker rats. Chloral hydrate (400 mg/kg) was used as the anesthetic agent. The in situ brain-pancreas perfusion model with intact pancreatic innervation was used in this investigation. The study measured insulin secretion in response to a 60-minute glucose stimulus (200 mg/dl). CNS-intact and CNS-functionally ablated obese and lean rats were used. During the 60-minute perfusion period significantly more insulin was released by pancreata from obese rats compared to those from lean rats. In lean rats, about twice as much insulin was released by pancreata from CNS-ablated rats than from CNS-intact rats (P < 0.05), demonstrating a CNS tonic inhibition of insulin secretion. In obese rats, there was no significant difference in insulin released by the pancreata of the CNS-intact and CNS-ablated rats. To determine if there was a masking effect of predominant PNS activity over the SNS in the CNS-intact obese rats, bilateral vagotomy was performed in a group of otherwise CNS-intact obese rats prior to the onset of perfusion. Tonic inhibition was still not observed in the CNS-vagotomized obese rats. In conclusion, hypersecretion of insulin in obese rats is partially due to diminished tonic sympathetic nervous system inhibition of insulin release. These results provide additional evidence regarding abnormal CNS control of insulin secretion in obese Zucker rats.