-1::1
Simple Hit Counter
Skip to content

Products

Solutions

×
×
Sign In

DE

EN - EnglishCN - 简体中文DE - DeutschES - EspañolKR - 한국어IT - ItalianoFR - FrançaisPT - Português do BrasilPL - PolskiHE - עִבְרִיתRU - РусскийJA - 日本語TR - TürkçeAR - العربية
Sign In Start Free Trial

RESEARCH

JoVE Journal

Peer reviewed scientific video journal

Behavior
Biochemistry
Bioengineering
Biology
Cancer Research
Chemistry
Developmental Biology
View All
JoVE Encyclopedia of Experiments

Video encyclopedia of advanced research methods

Biological Techniques
Biology
Cancer Research
Immunology
Neuroscience
Microbiology
JoVE Visualize

Visualizing science through experiment videos

EDUCATION

JoVE Core

Video textbooks for undergraduate courses

Analytical Chemistry
Anatomy and Physiology
Biology
Calculus
Cell Biology
Chemistry
Civil Engineering
Electrical Engineering
View All
JoVE Science Education

Visual demonstrations of key scientific experiments

Advanced Biology
Basic Biology
Chemistry
View All
JoVE Lab Manual

Videos of experiments for undergraduate lab courses

Biology
Chemistry

BUSINESS

JoVE Business

Video textbooks for business education

Accounting
Finance
Macroeconomics
Marketing
Microeconomics

OTHERS

JoVE Quiz

Interactive video based quizzes for formative assessments

Authors

Teaching Faculty

Librarians

K12 Schools

Biopharma

Products

RESEARCH

JoVE Journal

Peer reviewed scientific video journal

JoVE Encyclopedia of Experiments

Video encyclopedia of advanced research methods

JoVE Visualize

Visualizing science through experiment videos

EDUCATION

JoVE Core

Video textbooks for undergraduates

JoVE Science Education

Visual demonstrations of key scientific experiments

JoVE Lab Manual

Videos of experiments for undergraduate lab courses

BUSINESS

JoVE Business

Video textbooks for business education

OTHERS

JoVE Quiz

Interactive video based quizzes for formative assessments

Solutions

Authors
Teaching Faculty
Librarians
K12 Schools
Biopharma

Language

German

EN

English

CN

简体中文

DE

Deutsch

ES

Español

KR

한국어

IT

Italiano

FR

Français

PT

Português do Brasil

PL

Polski

HE

עִבְרִית

RU

Русский

JA

日本語

TR

Türkçe

AR

العربية

    Menu

    JoVE Journal

    Behavior

    Biochemistry

    Bioengineering

    Biology

    Cancer Research

    Chemistry

    Developmental Biology

    Engineering

    Environment

    Genetics

    Immunology and Infection

    Medicine

    Neuroscience

    Menu

    JoVE Encyclopedia of Experiments

    Biological Techniques

    Biology

    Cancer Research

    Immunology

    Neuroscience

    Microbiology

    Menu

    JoVE Core

    Analytical Chemistry

    Anatomy and Physiology

    Biology

    Calculus

    Cell Biology

    Chemistry

    Civil Engineering

    Electrical Engineering

    Introduction to Psychology

    Mechanical Engineering

    Medical-Surgical Nursing

    View All

    Menu

    JoVE Science Education

    Advanced Biology

    Basic Biology

    Chemistry

    Clinical Skills

    Engineering

    Environmental Sciences

    Physics

    Psychology

    View All

    Menu

    JoVE Lab Manual

    Biology

    Chemistry

    Menu

    JoVE Business

    Accounting

    Finance

    Macroeconomics

    Marketing

    Microeconomics

Start Free Trial
Loading...
Home
JoVE Journal
Biology
Manipulation und Analyse von zellzyklusabhängigen Prozessen in knospenden Hefen
Manipulation und Analyse von zellzyklusabhängigen Prozessen in knospenden Hefen
JoVE Journal
Biology
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Biology
Manipulation and Analysis of Cell Cycle-Dependent Processes in Budding Yeast

Manipulation und Analyse von zellzyklusabhängigen Prozessen in knospenden Hefen

Full Text
636 Views
08:13 min
September 26, 2025

DOI: 10.3791/68887-v

Michael G. Stewart*1,2, Talia C. Scheel*1, Ahmed A. Abouelghar*1, Sara E. Hoppe*1, Matthew P. Miller1

1Department of Biochemistry,University of Utah School of Medicine, 2Department of Molecular Biology and Genetics, Howard Hughes Medical Institute,Johns Hopkins University School of Medicine

AI Banner

Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This study investigates how yeast (S. cerevisiae) cells manage chromosome segregation during mitosis, utilizing synchronized cell cycles to observe cellular dynamics. By employing alpha-factor arrest in BAR1 mutants, researchers can achieve a precise G1 arrest to monitor changes in protein localization and activity throughout the cell cycle.

Key Study Components

Research Area

  • Cell cycle regulation
  • Chromosome segregation
  • Microscopy techniques

Background

  • Synchronized cell cycles unveil molecular processes otherwise hidden in unsynchronized populations.
  • Protein localization changes throughout the cell cycle are crucial for understanding mitosis.
  • Alpha-factor arrest provides a cleaner synchronization method than alternatives.

Methods Used

  • Fluorescence microscopy for imaging protein localization
  • Saccharomyces cerevisiae as the biological model
  • Alpha-factor treatment for G1 synchronization and subsequent release techniques

Main Results

  • Dynamic changes in protein localization were observed, particularly for Stu2-GFP during mitosis.
  • A peak in binucleate cells was noted at approximately 90 minutes post-release, indicating synchronized progress into anaphase.
  • Quantified intensity of protein puncta revealed significant changes correlating with different cell cycle stages.

Conclusions

  • The study effectively demonstrates precise methods for synchronizing yeast cell cycles to investigate mitotic processes.
  • This research has implications for broader biological understanding of cell division and chromosome behavior.

Frequently Asked Questions

What is the significance of studying yeast cells in cell cycle research?
Yeast cells serve as a simple eukaryotic model to study fundamental cell cycle processes that are conserved across species.
How does alpha-factor synchronization improve experiments?
Alpha-factor synchronization provides a precise and reversible means to arrest cells in G1 phase, allowing for controlled studies of the cell cycle.
What techniques are used to visualize protein localization?
Fluorescence microscopy techniques are employed to observe dynamic protein localization changes in real-time during the cell cycle.
What roles do Stu2-GFP and Spc110-mCherry play?
Stu2-GFP is used to track spindle dynamics, while Spc110-mCherry marks spindle pole bodies critical for mitotic spindle formation.
Why is synchronized cell population analysis important?
A synchronized cell population allows researchers to accurately assess changes in cellular behavior and protein dynamics at specific time points during the cell cycle.
What outcomes can one expect from this study?
The study aims to provide insights into the mechanisms underlying chromosome segregation and the dynamics of proteins involved in mitosis.
What potential applications does this research have?
Findings could inform cancer research and the development of therapeutic strategies targeting cell division processes.

Dieses Protokoll beschreibt zwei Methoden des Hefezellzyklus-Arrests und der optionalen Freisetzung und erläutert den Einsatz von Fluoreszenzmikroskopie zur Untersuchung von zellzyklusabhängigen Prozessen in S. cerevisiae.

Wir untersuchen, wie sich teilende Zellen während der Mitose ihre Chromosomen treu weitergeben, wobei wir uns auf die molekularen Maschinen und Mechanismen konzentrieren, die eine genaue Chromosomentrennung gewährleisten. Wir synchronisieren Zellen, um molekulare Prozesse zu untersuchen, die sich mit dem Zellzyklus verändern. Ohne diese Methoden wären wichtige Änderungen in einer unsynchronisierten Zellpopulation verborgen.

Im Vergleich zu anderen Synchronisationsmethoden bietet der Alpha-Faktor-Arrest bei BAR1-Mutanten einen saubereren, reversiblen G1-Arrest, der es uns ermöglicht, eine gesamte Hefekultur synchron im Zyklus zu verfolgen. Unsere Arbeit zeigt dynamische Veränderungen in der Proteinlokalisierung und -aktivität während des Zellzyklus auf und beleuchtet wichtige mitotische Prozesse wie Chromosomensegregation und Spindelpflege. Zu Beginn inokulieren Sie die Hefe in 25 Milliliter YPAD-Medium und inkubieren Sie über Nacht, um eine optische Dichte von 600 Nanometern zwischen 0,5 und 2,0 zu erreichen.

View the full transcript and gain access to thousands of scientific videos

View the full transcript and gain access to thousands of scientific videos

Sign In Start Free Trial

Explore More Videos

Diesen Monat in JoVE Ausgabe 223

Related Videos

Die Analyse der Entwicklung eines morphologischen Phänotyp als Funktion der Protein-Konzentration in Hefezellen

15:02

Die Analyse der Entwicklung eines morphologischen Phänotyp als Funktion der Protein-Konzentration in Hefezellen

Related Videos

11.3K Views

Analyse des Zellzyklus Position in Säugerzellen

12:19

Analyse des Zellzyklus Position in Säugerzellen

Related Videos

61.5K Views

Acquiring Fluoreszenz Zeitraffer-Filme von Bäckerhefe und Analysieren von Einzel-Zell-Dynamics mit Transplantaten

17:01

Acquiring Fluoreszenz Zeitraffer-Filme von Bäckerhefe und Analysieren von Einzel-Zell-Dynamics mit Transplantaten

Related Videos

13.4K Views

Räumlich-zeitliche Analyse von zytokinetischen Events in Spalthefe

11:19

Räumlich-zeitliche Analyse von zytokinetischen Events in Spalthefe

Related Videos

7.7K Views

Hochauflösende Bildgebung und Analyse einzelner Astral Dynamik der Mikrotubuli in Bäckerhefe

10:23

Hochauflösende Bildgebung und Analyse einzelner Astral Dynamik der Mikrotubuli in Bäckerhefe

Related Videos

10K Views

Inherent Dynamics Visualizer, eine interaktive Anwendung zur Bewertung und Visualisierung von Ergebnissen aus einer Gene Regulatory Network Inference Pipeline

10:44

Inherent Dynamics Visualizer, eine interaktive Anwendung zur Bewertung und Visualisierung von Ergebnissen aus einer Gene Regulatory Network Inference Pipeline

Related Videos

2.7K Views

Einsatz von Zeitraffer-Mikroskopie und stadienspezifischer Kerndepletion von Proteinen zur Untersuchung der Meiose bei S. cerevisiae

07:48

Einsatz von Zeitraffer-Mikroskopie und stadienspezifischer Kerndepletion von Proteinen zur Untersuchung der Meiose bei S. cerevisiae

Related Videos

2.3K Views

Abgleich von synchronisierten Zeitreihendaten unter Verwendung des Modells zur Charakterisierung des Zellzyklusverlusts für experimentübergreifende Vergleiche

07:59

Abgleich von synchronisierten Zeitreihendaten unter Verwendung des Modells zur Charakterisierung des Zellzyklusverlusts für experimentübergreifende Vergleiche

Related Videos

2K Views

Mess-replikative Lebensspanne in der Hefe

12:41

Mess-replikative Lebensspanne in der Hefe

Related Videos

21.4K Views

Quantitative Live Cell Fluoreszenz-Mikroskopie Analyse von Fission Yeast

06:52

Quantitative Live Cell Fluoreszenz-Mikroskopie Analyse von Fission Yeast

Related Videos

20.9K Views

JoVE logo
Contact Us Recommend to Library
Research
  • JoVE Journal
  • JoVE Encyclopedia of Experiments
  • JoVE Visualize
Business
  • JoVE Business
Education
  • JoVE Core
  • JoVE Science Education
  • JoVE Lab Manual
  • JoVE Quizzes
Solutions
  • Authors
  • Teaching Faculty
  • Librarians
  • K12 Schools
  • Biopharma
About JoVE
  • Overview
  • Leadership
Others
  • JoVE Newsletters
  • JoVE Help Center
  • Blogs
  • JoVE Newsroom
  • Site Maps
Contact Us Recommend to Library
JoVE logo

Copyright © 2026 MyJoVE Corporation. All rights reserved

Privacy Terms of Use Policies
WeChat QR code