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University of Turku

6 ARTICLES PUBLISHED IN JoVE

Developmental Biology

Analysis of Epididymal Protein Synthesis and Secretion

Wei Zhou^1,2, Petra Sipilä³, Geoffry N. De Iuliis^1,2, Matthew D. Dun^2,4, Brett Nixon^1,2

¹Priority Research Centre for Reproductive Science, School of Environmental and Life Sciences, University of Newcastle, ²Hunter Medical Research Institute, ³Department of Physiology, Turku Center for Disease Modeling, Institute of Biomedicine, University of Turku, ⁴School of Biomedical Sciences and Pharmacy, Faculty of Health and Medicine, University of Newcastle

Here, we report the immunofluorescence localization of dynamin to illustrate the protocols for the detection of proteins in paraffin-embedded mouse epididymal sections and those of an immortalized epididymal cell line (mECap18). We also describe the protocols for the isolation of secretory proteins from both epididymal fluid and conditioned cell media.

Biochemistry

Analysis of Thylakoid Membrane Protein Complexes by Blue Native Gel Electrophoresis

Marjaana Rantala¹, Virpi Paakkarinen¹, Eva-Mari Aro¹

¹Molecular Plant Biology, Department of Biochemistry, University of Turku

A protocol for the elucidation of plant thylakoid protein complex organization and composition with blue native polyacrylamide gel electrophoresis (BN-PAGE) and 2D-SDS-PAGE is described. The protocol is optimized for Arabidopsis thaliana, but can be used for other plant species with minor modifications.

Developmental Biology

Enrichment of Pachytene Spermatocytes and Spermatids from Mouse Testes Using Standard Laboratory Equipment

Matteo Da Ros^*1, Tiina Lehtiniemi^*1, Opeyemi Olotu¹, Oliver Meikar¹, Noora Kotaja¹

¹Institute of Biomedicine, University of Turku

Presented here is a protocol for enriching pachytene spermatocytes, round spermatids, and elongating spermatids from adult mouse testes using a discontinuous bovine serum albumin density gradient with standard laboratory equipment.

Biochemistry

Deciphering the Structural Effects of Activating EGFR Somatic Mutations with Molecular Dynamics Simulation

Mahlet Z. Tamirat¹, Kari J. Kurppa², Klaus Elenius^2,3,4, Mark S. Johnson¹

¹Structural Bioinformatics Laboratory, Biochemistry, Faculty of Science and Engineering, Åbo Akademi University, ²Medicity Research Laboratories and Institute of Biomedicine, University of Turku, ³Turku Bioscience Centre, University of Turku and Åbo Akademi University, ⁴Department of Oncology and Radiotherapy, University of Turku and Turku University Hospital

The objective of this protocol is to use molecular dynamics simulations to examine the dynamic structural changes that occur due to activating mutations of the EGFR kinase protein.

Developmental Biology

Transillumination-Assisted Dissection of Specific Stages of the Mouse Seminiferous Epithelial Cycle for Downstream Immunostaining Analyses

Juho-Antti Mäkelä¹, Sheyla Cisneros-Montalvo¹, Tiina Lehtiniemi¹, Opeyemi Olotu¹, Hue M. La^3,4, Jorma Toppari^1,2, Robin M. Hobbs^3,4, Martti Parvinen¹, Noora Kotaja¹

¹Institute of Biomedicine, Integrative Physiology and Pharmacology Unit, University of Turku, Turku, Finland, ²Department of Pediatrics, Turku University Hospital, Turku, Finland, ³Centre for Reproductive Health, Hudson Institute of Medical Research, Melbourne, VIC 3168, Australia, ⁴Australian Regenerative Medicine Institute, Monash University, Melbourne, VIC 3800, Australia

This protocol describes transillumination-assisted microdissection of segments of adult mouse seminiferous tubules representing specific stages of seminiferous epithelial cycle, and cell types therein, and subsequent immunostaining of squash preparations and intact tubule segments.

Environment

Quantification of the Potential Impact of Glyphosate-Based Products on Microbiomes

Suni Anie Mathew¹, Anne Muola¹, Kari Saikkonen¹, Irma Saloniemi², Marjo Helander², Pere Puigbò^2,3,4

¹Biodiversity Unit, University of Turku, ²Department of Biology, University of Turku, ³Nutrition and Health Unit, Eurecat Technology Centre of Catalonia, ⁴Department of Biochemistry and Biotechnology, Rovira i Virgili University

Glyphosate-based products (GBP) are the most common broad-spectrum herbicides worldwide. In this article, we introduce general guidelines to quantify the effect of GBP on microbiomes, from field experiments to bioinformatics analyses.