Current protocols in protein science 

About: Current protocols in protein science is an academic journal. The journal publishes majorly in the area(s): Fusion protein & Membrane protein. It has an ISSN identifier of 1934-3655. Over the lifetime, 1005 publications have been published receiving 29383 citations.

Comparative Protein Structure Modeling Using MODELLER

Abstract: Functional characterization of a protein sequence is a common goal in biology, and is usually facilitated by having an accurate three-dimensional (3-D) structure of the studied protein. In the absence of an experimentally determined structure, comparative or homology modeling can sometimes provide a useful 3-D model for a protein that is related to at least one known protein structure. Comparative modeling predicts the 3-D structure of a given protein sequence (target) based primarily on its alignment to one or more proteins of known structure (templates). The prediction process consists of fold assignment, target-template alignment, model building, and model evaluation. This unit describes how to calculate comparative models using the program MODELLER and discusses all four steps of comparative modeling, frequently observed errors, and some applications. Modeling lactate dehydrogenase from Trichomonas vaginalis (TvLDH) is described as an example. The download and installation of the MODELLER software is also described.

Assessing Reproductive Status/Stages in Mice

Abstract: The short reproductive cycle length observed in rodents, called the estrous cycle, makes them an ideal animal model for investigation of changes that occur during the reproductive cycle Most of the data in the literature about the estrous cycle is obtained from rats because they are easily manipulated and they exhibit a clear and well-defined estrous cycle However, the increased number of experiments using knockout mice requires identification of their estrous cycle as well, since (in)fertility issues may arise In mice, like rats, the identification of the stage of estrous cycle is based on the proportion of cell types observed in the vaginal secretion The aim of this unit is to provide guidelines for quickly and accurately determining estrous cycle phases in mice

Rodent models of depression: forced swim and tail suspension behavioral despair tests in rats and mice.

Abstract: The development of antidepressants requires simple rodent behavioral tests for initial screening before undertaking more complex preclinical tests and clinical evaluation. Presented in the unit are two widely used screening tests used for antidepressants, the forced swim (also termed behavioral despair) test in the rat and mouse, and the tail suspension test in the mouse. These tests have good predictive validity and allow rapid and economical detection of substances with potential antidepressant-like activity. The behavioral despair and the tail suspension tests are based on the same principle: measurement of the duration of immobility when rodents are exposed to an inescapable situation. The majority of clinically used antidepressants decrease the duration of immobility. Antidepressants also increase the latency to immobility, and this additional measure can increase the sensitivity of the behavioral despair test in the mouse for certain classes of antidepressant. Testing of new substances in the behavioral despair and tail suspension tests allows a simple assessment of their potential antidepressant activity by the measurement of their effect on immobility.

Simple behavioral assessment of mouse olfaction.

Abstract: This unit presents two basic protocols that offer rapid assessments of anosmia (the absence of a sense of smell) in mice The buried food test is used to check for the ability to smell volatile odors The olfactory habituation/dishabituation test is used to test whether the animal can detect and differentiate different odors, including both nonsocial and social odors A non-contact method of odor presentation, along with a general method for collecting urine samples, is given as an alternate protocol The tests described in this unit only require simple equipment and can be adopted readily by most laboratories

Automated Three-Chambered Social Approach Task for Mice

Abstract: Autism is diagnosed by three major symptom categories: unusual reciprocal social interactions, impaired communication, and repetitive behaviors with restricted interests. Direct social approach in mice has strong face validity to simple social approach behaviors in humans, which are frequently impaired in autism. This unit presents a basic protocol for a standardized, high-throughput social approach test for assaying mouse sociability. Our automated three-chambered social approach task quantifies direct social approach behaviors when a subject mouse is presented with the choice of spending time with either a novel mouse or a novel object. Sociability is defined as the subject mouse spending more time in the chamber containing the novel target mouse than in the chamber containing the inanimate novel object. The Basic Protocol describes procedures for testing one subject at a time in a single apparatus. A Support Protocol addresses data collection.