Cell Biology Laboratory Manual Table of Contents

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The Cell Cycle

We commonly think of dividing cells passing through a series of four stages, called the cell cycle, as they undergo sequential rounds of proliferation.  These four stages are:

 

           G1 - Pre-synthesis Gap.  Day to day function of the cell,

           S - Synthesis.  DNA replication

           G2 - Post-synthesis Gap.  Preparation for cell division

           M - Mitosis or cell division

 

When our focus is on mitosis, we may consider G1, S and G2 as a single unit called Interphase.  Although very different events are happening in the cell during these three stages, there is little to differentiate them microscopically.

 

S Stage

Before shifting our focus to mitosis, we do need to make sure we understand what is happening in S stage.   During S, the DNA is replicated making two identical copies of each double stranded DNA molecule.  The two daughter strands, however, remain attached by a centromere making a single large structure.  Because of this attachment, even though the amount of DNA in the cell has doubled, the number of chromosomes remains the same.  Each chromosome now has two identical chromatids attached by the centromere.

 

Interphase and Mitosis

 

When you examine a population of rapidly dividing cells, such as those at the tip of an onion, you will be able to see cells that are in Interphase or one of the four stages of mitosis.  Following are descriptions of cells in these various stages and brief descriptions of what is happening in the cell to cause the changes that you are seeing.

 

Interphase

Distinct nucleus.  Although the nuclear membrane is too thin to see with a compound microscope, you should be able to easily distinguish the nucleus (stained blue) from the more pinkish color of the cytoplasm.  The colors you observe are due to the nucleic acids staining blue while proteins stain red

Distinct nucleolus.  There are high levels of transcription going on in Interphase cells, especially of rRNA and tRNA.  The high concentration of nucleic acids make this portion of the nucleus stain more darkly.

Chromatin DNA.  When chromosomes are not condensed, it is called chromatin.  Because chromatin is very thin, you can not see individual DNA strands.  Therefore the blue of the nucleus is due to indistinct swirls of chromatin dispersed throughout the nucleus.

Prophase

In early prophase, the chromosomes will appear as thin strands dispersed about the entire nucleus.  The spindle apparatus will be forming but the thinness of the individual microtubule will probably make the spindle impossible to see at this early stage.

By middle prophase, the chromosomes strands will get thicker and shorter.  The nucleolus will be gone as transcription must stop as the DNA condenses.

In late prophase, the chromosomes will be very short and will be clearly moving towards the center of the spindle apparatus.

Metaphase

Short,  thick chromosomes,  The chromosomes reach their maximum level of condensation.

Chromosomes align on the metaphase plate.  The chromosomes are lined up in the middle of the spindle apparatus.  Although the chromatids may be pointing in various directions, the centromeres will be clearly all in a row.

Anaphase

Centromeres break.  The centromeres break releasing the sister chromatids as individual chromosomes.

Chromosomes separate.  Kinetochore microtubules shorten while polar microtubules elongate which pulls/pushes the chromosome towards the opposite poles of the spindle,  Recall that due to DNA replication in S phase that each new nucleus which forms will have precisely the same DNA content.

Telophase

Chromosome separation completed.  The chromosomes have reached opposite ends of the spindle.

Chromosomes unwind.  The chromosomes unwind and return to the chromatin conformation.

Cytokinesis.  You may be able to observe new cell wall forming in the middle of the old spindle apparatus.