I.

General information:

Every cell has (or has had during its development) a nucleus which contains almost all the genetic
information. It characterizes eukaryotic cells.

The nucleus or "vital center" of the cell is:

Limited by a nuclear envelope during interphase. Essential to the life of eukaryotic cells.

Carries the hereditary message in the form of DNA, which it preserves despite divisions thanks to
DNA replication.

Responsible for mRNA, tRNA and ribosomal RNA synthesis.

II. Structure :

A. Identification :

1. By standard staining: Haematoxylin-eosin, the nucleus appears basophilic.

2. By special staining: Toluidine blue, Feulgen stain.

B. Morphological characteristics of the nucleus :

The nucleus is bounded by a nuclear envelope formed by two membranes separated by a perinuclear
space, and contains :

 Nucleoplasm of low stainability.

 Clusters of a highly chromophilic substance, chromatin.

 Spherical bodies, the nucleoli.

-Size: Varies according to cell type. The Nucleo-Cytoplasmic Ratio (NCR) is the ratio of nucleus volume
to cell volume-nucleus volume.

This ratio is constant for each cell type and is species-specific.

-Shape: the shape of the nucleus differs according to the shape of the cell.
It may be rounded in cubic cells, ovoid in cylindrical cells, discoid or polylobed ....

-Position: the nucleus can be :

 Central: lymphocytes, fibroblasts, endocrine gland cells.

 Repressed at the base of the cell: mucous cells, exocrine gland cells.

 Peripheral: striated muscle cells, adipocytes.

-Number: most cells have a single nucleus, but there are exceptions:

-Red blood cells and keratinocytes (cells of the superficial layers of the epidermis) are anucleate cells.

-Hepatocytes (liver cells) usually have two or more nuclei.

-Osteoclasts, the giant cells of bone tissue, have an average of ten nuclei.

III. Ultrastructure :

The interphase nucleus under electron microscopy

Nucléoplasme
A. The nuclear envelope: a complex membrane assembly characteristic of eukaryotic cells:

 separates chromatin from hyaloplasm during interphase.

 And controls two-way exchanges between nucleus and hyaloplasm.

1. Ultra structure:

The nuclear envelope appears to consist of two tri-layered membranes, each 75A° thick.

These two membranes are separated by a peri-nuclear space 200 to 400A° wide.

a. The outer membrane :

- Is lined on its hyaloplasmic side with ribosomes, and is continuous with the endoplasmic reticulum.

- Contains 70% proteins and 30% lipids.

- Very rich in enzymes: glucose-6-phosphatase, two electron transport chains (cytochromes).

b. The perinuclear space: located between the two membranes, this is where calcium ions are
stored.

c. Inner membrane:

- The inner membrane faces the nucleoplasm, and is lined internally by the lamina.

- Structurally similar to the outer membrane, but with less enzymatic activity.

- Contains transmembrane proteins acting as binding sites for lamins and chromatin proteins
(histones).

- Transmembrane calcium channels, which release calcium ions contained in the perinuclear space.
Ex: Ca++ATPase.

d. Role of the nuclear envelope: it :

 Acts as a barrier controlling the passage of water, ions and macromolecules.

 Involved in nucleocytoplasmic exchanges.

 Participates in protein synthesis, through its ribosome-lined outer membrane.

 Actively transports Ca++ and stores it in the perinuclear space.

 Nuclear envelope (1)

B. The nuclear pore:

1. Definition :

Nuclear pores: are :

- Complex structures consisting of interrupted areas of the nuclear envelope,

- Formed by an assembly of positively-charged proteins (around fifty) called nucleoporins.

- With a molecular weight of around 125 million Daltons, they are involved in exchanges between the
nucleus and the cytoplasm.

2. Pore dynamics:

Pores are not permanent structures, but dynamic ones, likely to disappear during cell rest and
reappear when nucleocytoplasmic exchanges are increased.

The number of pores is around 3,000 to 4,000 per nucleus (5 to 15% of the envelope surface). It
varies according to the physiological state of the cell, particularly during cell growth (embryogenesis).
 3. three-dimensional structure:

It shows an organization into 8 subunits, these eight subunits form two rings, one cytoplasmic, the
other nucleoplasmic. Each ring carries perpendicular filaments, the filaments on the nucleoplasmic
side being connected at their ends to form a nuclear basket. The whole is anchored in the nuclear
envelope.

Towards the pore, each s/unit emits an arm, connected to the two rings. The arms leave thin lateral
tunnels of around 10nm between them. Together, the arms form a central tunnel of the order of
40nm. The central tunnel is closed by a diaphragm, opening as molecules pass through in the
presence of energy.

4. Role of the pore:

Pores control nucleocytoplasmic exchanges in both directions, cytoplasm-nucleus and nucleus-

cytoplasm, and enable selective transit:

- Small molecules (nucleotides, proteins...) of molecular weight < 4OKda and ions pass through the
pore without external intervention by passive diffusion: these exchanges use the lateral channels of
the pore.

- Large molecules are transported via the central transporter and sit on specialized transport
proteins, whose transport requires energy:

 Transport involves an addressing system based on the existence of specific amino acid sequences.
Only proteins carrying this signal will be transported.

 Transport involves an adaptor (importin, exportin) recognizing an addressing system.

Filaments Cytoplasmiques
Cross-section of a nuclear pore (2)
Le complexe du pore nucléaire

Vue de Face (4)

Transport through the pore (4)

D. The lamina: is a dense fibrillar protein network, closely linked to the inner surface of the
envelope. The lamina is made up of fibrous polypeptides known as lamins, cytoskeletal proteins of
the intermediate filament family (lamins A, B and C).

The lamina dissociates at the start of mitosis and finally reorganizes during cell division.

◾ Roles: forms a scaffold that gives the nucleus its shape and maintains the rigidity of the nuclear
envelope.

◾ Pathology: progeria:-is an extremely rare genetic disease characterized by premature aging.

- Due to a mutation in the gene that codes for lamins A located on chromosome 1.

E. Chromatin:

1. Definition: Chromatin is the content of nucleoplasm in the nuclei of cells in interphase

(outside mitosis), which can be observed by light or electron microscopy.

Chromatin comes in two forms:

- Heterochromatin: dark after staining under light microscopy, electron-dense under electron
microscopy; represents 80 to 90% of the total chromatin of an adult cell and corresponds to
metabolically inactive regions (no transcription).

- Euchromatin: clear after staining under light microscopy and less electron-dense under electron
microscopy; represents 10 to 20% of adult cell chromatin and corresponds to metabolically active
regions (transcription).

2. Distribution:

a) Euchromatin is distributed throughout the nucleoplasm.

b) Heterochromatin, on the other hand, is found

 At the periphery of the nucleus, pressed against the nuclear membrane where it forms the
chromatic membrane.

 In one or more juxta-nucleolar clumps.

3. Structure :

Light microscopy: appears identical for nuclei belonging to the same cell types, but varies from one
cell type to another depending on the cell's activity. It takes the form of :

- Small or large lumps.

- Granulations or sprays.

- Confluent mesh networks in places.

Barr's corpuscle (sex chromatin): a mass of heterochromatin found in the nuclei of female somatic
cells.

1um in diameter, corresponding to one of the X chromosomes (inactivated).

Electron microscopy: the ultra-structure of the chromatin shows

the presence of tightly-packed fibers with a spiral trajectory: chromatin fibers or nucleosomal fibers.
These fibers are made up of a succession of elementary structures, called nucleosomes, formed by a
histone octamer around which DNA is wound.

The state of chromatin condensation depends on the cell's activity. A cell undergoing intense activity
has a high level of decondensed (clear) chromatin.
 The nucleus under electron microscopy

4. Chemical composition: Chromatin is made up of DNA and associated proteins.

4. Chemical composition: Chromatin is made up of DNA and associated proteins: histones.

a) DNA: nuclear DNA contains the genetic heritage (information transmitted from generation to
generation) necessary for the synthesis of structural and enzymatic proteins.

Primary structure: the DNA molecule is formed by the association of two strands: a double-stranded
molecule. Each strand is a polymer, formed by the association of deoxyribonucleotides linked
together by phosphodiester bonds.

b) Histones: are basic proteins of five types: H1, H2A, H2B, H3 and H4.
 Structure de l’ADN

5. DNA replication: is the mechanism by which a cell copies its DNA in order to transmit the
same genetic information to its daughter cells after division. It takes place during the S phase, the
synthesis phase of the cell cycle.

F. The nucleolus:
1. General: The nucleolus:

Appears as a dense spherical corpuscle.

Considered a nuclear organelle, not bounded by a membrane. Visible under light and electron
microscopy.

Is a dynamic structure, present during interphase and disappearing during mitosis.

The number can vary from one to several per cell.

Size varies according to cell activity (protein synthesis).

Its main function is ribosome biogenesis,

2. Structure under light microscopy: under light microscopy, the nucleolus:

Appears as a spherical or oval refractive body, surrounded by a ring of chromatin.

It comprises:

-Perinucleolar chromatin: arranged in a crescent shape more or less completely surrounding the
nucleolar body.

-The nucleolar body: appears as a dense homogeneous structure, measuring 1 to 2um in normal
cells.

3. Components of the ME nucleolus: microscopy reveals the macromolecular constituents of

the nucleolus.

Three components have been identified:

1. Fibrillar centers (FC).

2. A dense fibrillar component (DFC), partially or totally surrounding the fibrillar centers.

3. A granular component (GC) in which the fibrillar centers and dense granular components are
embedded.

4. Organization of the nucleolus:

The nucleolus contains large loops of DNA, belonging to five pairs of acrocentric chromosomes13, 14,
15, 21, 22, each of which contains a group of rRNA genes about forty per chromosome. Each of these
groups constitutes a nucleolar organizing center or NOR.

The three nucleolar regions:

1. the fibrillar center: which contains proteins involved in transcription

such as RNA polymerase I and transcription factors.

2. Dense fibrillar component: which contains a protein called fibrillarin involved in rDNA
transcription and pre-rRNA cleavage.

3. Granular component: contains the pre-ribosomes.

Structure of the nucleolus using electron microscopy

5. Functions of the nucleolus:

a. Ribosome biosynthesis: which requires:

- Transcription of rDNA into 45S rRNA ;

- Maturation of neoformed 45S rRNA and its cleavage into 5,8S, 18S, 28S;

- Assembly into ribosomal subunits: -18S rRNA associates with imported ribosomal proteins to give
the small ribosomal subunit (40 S) and exits the nucleoplasm through the nuclear pores.

- rRNA 5, 8 and 28s associate with each other and with ribosomal proteins imported from the
cytoplasm and with RNA 5s (which is transcribed from extranuclear DNA by polymerase

III), to form the large ribosome subunit (60S), which then enters the cytoplasm through the nuclear
pores.

b. Cell cycle regulation: the nucleolus collaborates with regulatory proteins to control the cell
cycle. This role has been highlighted by the study of the control of cell cycle stalling following DNA
strand damage caused by physical (e.g. ionizing radiation) or chemical (e.g. pesticides) aggression.

6. Nucleoli and disease:

◾ Werner syndrome: is a hereditary disease characterized by premature aging, due to a mutation in a

protein involved in nucleolar DNA transcription.

◾ Treacher Collins syndrome: which is a genetic disease that affects the development of the face and
skull, due to a mutation in a gene that codes for a nucleolar protein (treacle).
13, 14, 15,21 et 22
 Noyau

Cytoplasme

Morphological characteristics of cancer cells:

Morphologically, cancer cells may show many modifications of normal cell characteristics, but no
cytological criteria are specific to cancer.

Intra-cellular organelles may be affected in size, shape or number.

number.

1- Nucleus abnormalities: the appearance of the nucleus is the most characteristic sign.

- Anisokaryosis: unequal size from one nucleus to another in the same tissue (monstrous nuclei).

- Irregularity of shape and contour.

- Multiple nuclei.

- Increased nucleocytoplasmic ratio.

- Chromatin condensation, or nuclear hyperchromatism.

- Nucleolus abnormalities: enlarged, irregular nucleolus, or multinucleation.

2- Mitosis abnormalities: the number and atypical nature of mitoses are indicative of malignancy.

- Increased number of mitoses with shortened interphase.

- Abnormal, multipolar mitoses with irregular distribution of chromosomes.

3- Cytoplasmic abnormalities :

- Anisocytosis: cells of irregular size and shape, cellular gigantism.

- Cytoplasm is reduced and basophilic.

- Accumulation of certain elaboration products e.g. keratin, mucus, glycogen.

4- Plasma membrane abnormalities :

- light microscopy: thickened and irregular.

- in electron microscopy: it is bristling with amorphous, irregular microvilli; cell junctions are also
affected.

References :
1. Cell Biology. Abrégés. Marc Maillet.9th edition, Masson2002.

2. Cell Biology. Y Bassaglia. Maloine 2001.

3. Cellular Biology. PCEM1. Ediscience .2007.

4. Cours de Biologie Cellulaire : Pierre Cau, Raymond Seite. Edition ellipses.1999.

5. Cytology & Cell Physiology. M. Abdelali, H. Benzine-Challam, A.Madoui-

6. Dekar. Office des Publications Universitaires 2008.

7. The cell and its physiology: M Bendjelloul. Office des Publications Universitaires 2011.

8. Mini manual de Biologie Cellulaire: cours QCM, QROC. J M Petit, S Arico, R Julien. Dumond
2008.