TY - JOUR AB - A better alignment of preclinical and clinical neurobiological measures could help improve neuropsychiatric disease therapeutic development. This unit describes a compendium of hypothesis-driven neuroanatomical phenotyping strategies to be employed in genetic mouse models. Using neuropsychiatric disease-based alterations as a guide, these are histological and immunohistochemical methodologies also applied to human tissue. They include quantification assays of neurochemical-, newly born neuron- and glial-cell markers, synaptic proteins, regional volumetrics, dendritic complexity and spine number as well as an index of excitation/inhibition balance. The techniques can be implemented in isolation or to complement concordant behavioral and electrophysiological analyses. Each outcome will provide functional detail necessary to decipher underlying neural circuit abnormalities associated with a brain-related phenotype in mice. Experimental design, timing, anticipated results and potential pitfalls are discussed. © 2018 by John Wiley & Sons, Inc. AU - Garrett, L. AU - Ung, M.-C. AU - Heermann, T. AU - Niedermeier, K.M. AU - Hölter, S.M. C1 - 53998 C2 - 45185 SP - 79-128 TI - Analysis of neuropsychiatric disease-related functional neuroanatomical markers in mice. JO - Curr. Protoc. Mouse Biol. VL - 8 IS - 1 PY - 2018 SN - 2161-2617 ER - TY - JOUR AB - The cell composition of bronchoalveolar lavage fluid (BAL) is an important indicator of airway inflammation. It is commonly determined by cytocentrifuging leukocytes on slides, then staining, identifying, and counting them as eosinophils, neutrophils, macrophages, or lymphocytes according to morphological criteria under light microscopy, where it is not always easy to distinguish macrophages from lymphocytes. We describe here a one-step, easy-to-use, and easy-to-customize 8-color flow cytometric method for performing differential cell count and comparing it to morphological counts on stained cytospins. This method identifies BAL cells by a simultaneous one-step immunolabeling procedure using antibodies to identify T cells, B cells, neutrophils, eosinophils, and macrophages. Morphological analysis of flow-sorted cell subsets is used to validate this protocol. An important advantage of this basic flow cytometry protocol is the ability to customize it by the addition of antibodies to study receptor expression at leukocyte cell surfaces and identify subclasses of inflammatory cells as needed. © 2017 by John Wiley & Sons, Inc. AU - Daubeuf, F.* AU - Becker, J.* AU - Aguilar-Pimentel, A. AU - Ebel, C.* AU - Hrabě de Angelis, M. AU - Hérault, Y.* AU - Frossard, N.* C1 - 51365 C2 - 43185 SP - 88-99 TI - A Fast, Easy, and Customizable Eight-Color Flow Cytometric Method for Analysis of the Cellular Content of Bronchoalveolar Lavage Fluid in the Mouse. JO - Curr. Protoc. Mouse Biol. VL - 7 IS - 2 PY - 2017 SN - 2161-2617 ER - TY - JOUR AB - Adaptive social behavior is important in mammals, both for the well-being of the individual and for the thriving of the species. Dysfunctions in social behavior occur in many neurodevelopmental and psychiatric diseases, and research into the genetic components of disease-relevant social deficits can open up new avenues for understanding the underlying biological mechanisms and therapeutic interventions. Genetically modified mouse models are particularly useful in this respect, and robust experimental protocols are needed to reliably assess relevant social behavior phenotypes. Here we describe in detail three protocols to quantitatively measure sociability, one of the most frequently investigated social behavior phenotypes in mice, using a three-chamber sociability test. These protocols can be extended to also assess social memory. In addition, we provide a detailed protocol on pup retrieval, which is a particularly robust maternal behavior amenable to various scientific questions. © 2017 by John Wiley & Sons, Inc. AU - Zimprich, A. AU - Niessing, J.* AU - Cohen, L.* AU - Garrett, L. AU - Einicke, J. AU - Sperling, B. AU - Schmidt, M.V.* AU - Hölter, S.M. C1 - 52580 C2 - 44133 SP - 287-305 TI - Assessing sociability, social memory, and pup retrieval in mice. JO - Curr. Protoc. Mouse Biol. VL - 7 IS - 7 PY - 2017 SN - 2161-2617 ER - TY - JOUR AB - Phenotyping of inbred mouse strains and genetically modified mouse models for characteristics related to neuropsychiatric diseases includes assessing their anxiety-related behavior. A variety of tests have been developed to measure anxiety in laboratory rodents and these tests have been placed under scrutiny over the years concerning their validity. Here we describe the most widely used tests for anxiety in mice. The protocols we present are established methods used in the German Mouse Clinic (GMC), with which alterations in anxiety could successfully be discovered in mouse mutants. Moreover, since baseline anxiety levels in mice are easily influenced by a great variety of disturbances, we carefully outline the critical parameters that need to be considered. AU - Hölter, S.M. AU - Einicke, J. AU - Sperling, B. AU - Zimprich, A. AU - Garrett, L. AU - Fuchs, H. AU - Gailus-Durner, V. AU - Hrabě de Angelis, M. AU - Wurst, W. C1 - 47502 C2 - 39340 SP - 291-301 TI - Tests for anxiety-related behavior in mice. JO - Curr. Protoc. Mouse Biol. VL - 5 IS - 5 PY - 2015 SN - 2161-2617 ER - TY - JOUR AB - Genetically modified mouse models have proven useful to study learning and memory processes and the neurocircuitry and molecular mechanisms involved, as well as to develop therapies for diseases involving cognitive impairment. A variety of tests have been developed to measure cognition in mice, and here we present those established and regularly used in the German Mouse Clinic. The test paradigms have been carefully chosen according to reliability of results and disease relevance of the cognitive functions assessed. Further criteria were time efficiency and ease of application. All tests assess slightly different but also overlapping or interacting aspects of learning and memory so that they can be used to complement each other in a comprehensive assessment of cognitive function. The five protocols described are for spontaneous alternation in the Y-maze, social discrimination, object recognition, automated assessment of learning and memory using the IntelliCage, and olfactory discrimination learning. AU - Hölter, S.M. AU - Garrett, L. AU - Einicke, J. AU - Sperling, B. AU - Dirscherl, P. AU - Zimprich, A. AU - Fuchs, H. AU - Gailus-Durner, V. AU - Hrabě de Angelis, M. AU - Wurst, W. C1 - 47503 C2 - 39339 SP - 331-358 TI - Assessing cognition in mice. JO - Curr. Protoc. Mouse Biol. VL - 5 IS - 5 PY - 2015 SN - 2161-2617 ER - TY - JOUR AB - Current comprehensive mouse metabolic phenotyping involves studying energy balance in cohorts of mice via indirect calorimetry, which determines heat release from changes in respiratory air composition. Here, we describe the measurement of daily energy expenditure (DEE) and basal metabolic rate (BMR) in mice. These well-defined metabolic descriptors serve as meaningful first-line read-outs for metabolic phenotyping and should be reported when exploring energy expenditure in mice. For further guidance, the issue of appropriate sample sizes and the frequency of sampling of metabolic measurements is also discussed. AU - Meyer, C.W. AU - Reitmeir, P. AU - Tschöp, M.H. C1 - 46724 C2 - 37768 SP - 205-222 TI - Exploration of energy metabolism in the mouse using indirect calorimetry: Measurement of Daily Energy Expenditure (DEE) and Basal Metabolic Rate (BMR). JO - Curr. Protoc. Mouse Biol. VL - 5 IS - 3 PY - 2015 SN - 2161-2617 ER - TY - JOUR AB - This article presents a detailed description of intraperitoneal and oral glucose tolerance tests in mice. The former is widely used in initial high-throughput phenotyping of mutant mice to assess a diabetic phenotype and alterations in glucose homeostasis. Each protocol provides a comprehensive description of each step in the workflow, including variation of the standard protocol under particular circumstances (e.g., sensitivity to food deprivation, excessive deviations in body composition, or need for extra blood samples for additional analyses). We also describe how reduction of body mass and body temperature can be used as additional readouts to monitor metabolic function in response to food deprivation. AU - Rozman, J. AU - Rathkolb, B. AU - Neschen, S. AU - Fuchs, H. AU - Gailus-Durner, V. AU - Klingenspor, M.* AU - Wolf, E.* AU - Hrabě de Angelis, M. C1 - 43548 C2 - 36562 SP - 65-84 TI - Glucose tolerance tests for systematic screening of glucose homeostasis in mice. JO - Curr. Protoc. Mouse Biol. VL - 5 IS - 1 PY - 2015 SN - 2161-2617 ER - TY - JOUR AB - Historically, timed mating of either naturally cycling or superovulated females has been the mainstay of pre-implantation embryo production. However, rising cage costs and the rapid expansion of biomedical research programs has necessitated the development of high-throughput approaches to mouse embryo production. In vitro fertilization (IVF) represents one such versatile tool offering many advantages to busy mouse facilities in terms of efficient use of space and resources. For example, strains can be taken off the shelf, frozen quickly as sperm, and recovered at a later date, small colonies can be rapidly expanded to meet demand, and IVF can be used to rescue strains that fail to breed or where the founder male is ill or has died suddenly. This article describes an IVF protocol currently used by many reproductive technologists to assist mouse biology programs. AU - Guan, M.* AU - Bogani, D.* AU - Marschall, S. AU - Raspa, M.* AU - Takeo, T.* AU - Nakagata, N.* AU - Fray, M.* C1 - 44451 C2 - 36850 SP - 67-83 TI - In vitro fertilization in mice using the MBCD-GSH protocol. JO - Curr. Protoc. Mouse Biol. VL - 4 IS - 2 PY - 2014 SN - 2161-2617 ER - TY - JOUR AB - Each year, thousands of new mouse models are generated around the world to further biomedical research. Unfortunately, the cost of maintaining mouse colonies makes it uneconomical to keep strains on the shelf that are not part of active research programs. Ideally, these retired strains should be archived. If this is not done and the line is simply killed off, the genetics are lost to future generations of scientists. Traditionally, embryo freezing has been used to cryopreserve mice, but this is expensive, time consuming, requires large numbers of donor females, and usually involves invasive superovulation procedures. Sperm freezing circumvents all of these disadvantages and is rapidly becoming the technique of choice for many repositories. This has been made possible through the use of refined cryoprotective agents and the development of improved in vitro fertilization techniques. This article describes two popular sperm freezing techniques employed by mouse repositories to archive spermatozoa using cryoprotective agents supplemented with either L-glutamine or monothioglycerol. AU - Guan, M.* AU - Bogani, D.* AU - Marschall, S. AU - Raspa, M.* AU - Takeo, T.* AU - Nakagata, N.* AU - Taft, R.* AU - Fray, M.* C1 - 44452 C2 - 36851 SP - 85-104 TI - Contemporary techniques for freezing mouse spermatozoa. JO - Curr. Protoc. Mouse Biol. VL - 4 IS - 3 PY - 2014 SN - 2161-2617 ER - TY - JOUR AB - The ability to interrogate the entire coding sequence of the mouse combined with the tools to manipulate the genome has firmly established the mouse as the model organism of choice for studying the causes of human disease. Consequently, a huge number of novel mouse models are generated each year to support active research programs. However, it is neither ethically justifiable, nor economically viable to maintain mouse colonies on the shelf that are not part of active research programs. This means that novel mouse lines have to be preserved in some way. If this is not done and the line is simply killed off, the genetics will be lost to future generations of scientists. This article describes the current practices used in cryopreservation laboratories to archive and recover mouse embryos frozen using controlled-rate freezing and vitrification techniques. AU - Guan, M.* AU - Bogani, D.* AU - Marschall, S. AU - Raspa, M.* AU - Takeo, T.* AU - Nakagata, N.* AU - Fray, M.* C1 - 44454 C2 - 36853 SP - 205-227 TI - Conservation of mouse models through embryo freezing. JO - Curr. Protoc. Mouse Biol. VL - 4 IS - 4 PY - 2014 SN - 2161-2617 ER - TY - JOUR AB - The 21st century has seen a huge proliferation in the availability of genetically altered mice. The availability of these resources has been accompanied by ever greater opportunities for international collaborations between laboratories involving the exchange of mouse strains. This exchange can involve significant costs in terms of animal welfare and transportation expenses. In an attempt to mitigate some of these costs, the mouse community has developed a battery of techniques that can be used to avoid transporting live mice. Transporting frozen embryos and sperm at liquid nitrogen (LN2 ) temperatures using dry shippers has been common practice for some time. However, current advances in this field have refined transportation procedures and introduced new techniques for disseminating embryos and sperm: for example, shipping frozen sperm on dry ice, exchanging unfrozen epididymides from which sperm can be extracted, and transporting frozen/thawed embryos in isotonic media. This article discusses some of the current practices used by laboratories to transport mouse strains around the world without having to exchange live mice. AU - Kenyon, J.* AU - Guan, M.* AU - Bogani, D.* AU - Marschall, S. AU - Raspa, M.* AU - Pickard, A.* AU - Takeo, T.* AU - Nakagata, N.* AU - Fray, M.* C1 - 44453 C2 - 36852 SP - 47-65 TI - Transporting mouse embryos and germplasm as frozen or unfrozen materials. JO - Curr. Protoc. Mouse Biol. VL - 4 IS - 2 PY - 2014 SN - 2161-2617 ER - TY - JOUR AB - Basic phenotyping of inbred mouse strains and genetically modified mouse models usually includes the determination of blood-based parameters as a diagnostic screen for genotype effects on metabolism and organ function. A broad range of analytes, including hematological parameters, can be reliably determined in mouse blood, if appropriate samples are available. Here we describe recommended techniques for blood collection from mice and the considerations that have to be taken into account to get adequate samples for hematological investigations. Furthermore, we describe established methods used in the German Mouse Clinic (GMC) to determine hematological parameters in the mouse. AU - Rathkolb, B. AU - Fuchs, H.* AU - Gailus-Durner, V. AU - Aigner, B.* AU - Wolf, E.* AU - Hrabě de Angelis, M. C1 - 25476 C2 - 31854 SP - 101-119 TI - Blood collection from mice and hematological analyses on mouse blood. JO - Curr. Protoc. Mouse Biol. VL - 3 PB - Wiley-Blackwell PY - 2013 SN - 2161-2617 ER - TY - JOUR AB - Besides hematological analyses, many other parameters, including clinical chemistry and endocrinological values, can be determined from mouse blood samples. For most of these tests, plasma or serum samples are used. Data obtained by these investigations provide indications of genotype effects on metabolism and organ functions. Here we describe in detail the considerations that have to be taken into account to get adequate samples for plasma or serum analyses and the recommended sample processing for different investigations. Furthermore, we describe established methods used in the German Mouse Clinic (GMC) to determine clinical chemical parameters; for more in-depth analysis of specific classes of biomarkers, we provide instructions for ELISAs (sandwich and competitive) as well as LC-MS/MS, focusing on markers associated with bone or steroid metabolism in the mouse as working examples. AU - Rathkolb, B. AU - Hans, W. AU - Prehn, C. AU - Fuchs, H. AU - Gailus-Durner, V. AU - Aigner, B.* AU - Adamski, J. AU - Wolf, E.* AU - Hrabě de Angelis, M. C1 - 25475 C2 - 31855 SP - 69-100 TI - Clinical chemistry and other laboratory tests on mouse plasma and serum. JO - Curr. Protoc. Mouse Biol. VL - 3 PB - Wiley-Blackwell PY - 2013 SN - 2161-2617 ER -