Biotechnology Methods / Cell Biology and Genetics

Meiosis in Grasshopper Testis (Poecilocerus Pictus)

e testes of the grasshopper are removed and xed in Carnoy’s uid. A er 2–14 hours, the testes are transferred to 10% alcohol and
stored.

Squash Preparation
1. 1 or 2 lobes of the testes are removed.
2. e testes are placed on a glass slide.
3. Apply 1 to 2 drops of acetocarmine stain.
4. With a sharp blade, the teste lobes are cut into minute pieces and kept for
10 minutes.
5. e slide is then gently covered with a coverslip, taking care so that air
bubbles are not formed.
6. Warm the slide gently and place it between 2 folds of lter paper.
7. Press the material with the tip of the nger and remove the excess stain,
which comes out on the sides of the coverslip.
8. e slide is observed under the microscope.

Interphase
is phase is usually present in animal cells. e cells in this stage are physiologically active. No DNA replication takes place.

Prophase-I
a. Leptotene. e chromosomes are long, standard, and uncoiled. ey are densely formed on 1 side of the cell. Only 1 sex
chromosome occurs in the males, which normally replicates later and hence appears as a dark skin body.
b. Zygotene. Homologous chromosomes pair by a process called synapsis. Pairing starts from many points on the chromosome. e
chromosomes are called bivalents. Bivalents become shortened and thickened by coiling and condensation. Synapsis of a
chromosome is cemented by a complex called synaptonemal complex, which facilitates crossing over.
c. Pachytene. Crossing over takes place between nonsister chromatids. Crossing over is accompanied by the chiasmata formation.
d. Diplotene. Condensation of chromatid material is greater. Each chromosome can be distinguished separately.
e. Diakinesis. Homologous chromosomes begin to coil and become shorter and thicker. Chromosomes are fully contracted and
deeply stained. e ‘X’ chromosome is rod-shaped, univalent, and easily distinguishable from the rest of the chromosomes.

Metaphase-II
e chromosomes get oriented in the equatorial region of the spindle and their centromeres are attached to the chromosomal bers.
Each chromosome is easily seen. Maximum concentration occurs at this stage.

Anaphase-II
e spindle bers contract and the homologous chromosomes separate and move toward the opposite poles. Each chromosome
consists of 2 chromatids attached to 1 centromere.

Telophase-I
e separation of homologous chromosomes is completed. ey reach the opposite poles. Two distinct daughter nuclei are formed.
e daughter nuclei formed contain only half the number of chromosomes present in the parent cell. Cytokinesis may occur a er
the completion of telophase.

Prophase-II
e chromosomes with 2 chromatids become short and thick. is is the stage of the second meiotic division. e nuclear
membrane and the nucleolus are absent. e spindle is formed and the chromosomes are arranged on the equator.

Anaphase-II
e spindle is formed. e centromeres of the daughter chromosomes are attached to the spindle bers. e 2 groups of the
daughter chromosomes in each cell have started moving apart toward the opposite poles of the spindle
daughter chromosomes in each cell have started moving apart toward the opposite poles of the spindle.

Telophase-II
e 2 groups of daughter chromosomes in each haploid cell have reached the 2 poles of the spindle. e 2 haploid daughter cells
formed as a result of rst meiotic division divide again by the second meiotic division. Four haploid cells are formed from a single
diploid cell.

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