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Oran1 Ahmed Benbella University, Faculty of Medicine, Department of Medicine
Histology-Embryology Department, Dr Belarbi-Amar
N
Cytology class for first-year medical students
The interphasic core
I. General information:
Every cell has (or has had during its development) a nucleus that contains almost all its genetic
information. It characterizes eukaryotic cells.
The nucleus or "vital center" of the cell is:
Limited by a nuclear envelope during interphase. Essential
to eukaryotic cell life.
Carrier of the hereditary message in the form of DNA, which it retains despite
divisions thanks to DNA replication.
Responsible for mRNA, tRNA and ribosomal RNA synthesis.
II. Structure :
A. Highlight:
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 is not very stainable.
⮚ Clusters of a highly chromophilic substance, chromatin.
⮚ Spherical bodies, the nucleoli.
-Size: This varies according to cell type. The Nucleo-cytoplasmic ratio (RNP) is the
ratio: nucleus volume/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 can be rounded in cubic cells, ovoid in cylindrical cells, discoid or polylobed ....
-Position: the core can be :
1 Academic year: 2019-2020
Oran1 Ahmed Benbella University, Faculty of Medicine, Department of Medicine
Histology-Embryology Department, Dr Belarbi-Amar
N
Cytology class for first-year medical students
✓ Central: lymphocytes, fibroblasts, endocrine gland cells.
✓ Repressed at the base of the cell: mucous cells, exocrine glandular 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 :
Nucleoplasm
The interphase nucleus
under electron
microscopy
2 Academic year: 2019-2020
Oran1 Ahmed Benbella University, Faculty of Medicine, Department of Medicine
Histology-Embryology Department, Dr Belarbi-Amar
N
Cytology class for first-year medical students
A. 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 be formed by 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 :
• The hyaloplasmic surface is lined with ribosomes, and is continuous
with the endoplasmic reticulum.
• Contains 70% protein and 30% fat.
• 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. The 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 release calcium ions contained in
the perinuclear space. Ex: Ca++ATPase.
d. Role of the nuclear envelope: :
✓ Acts as a barrier controlling the passage of water, ions and macromolecules.
3 Academic year: 2019-2020
Oran1 Ahmed Benbella University, Faculty of Medicine, Department of Medicine
Histology-Embryology Department, Dr Belarbi-Amar
N
Cytology class for first-year medical students
✓ Involved in nucleocytoplasmic exchanges.
✓ Involved in protein synthesis, its outer membrane is lined with ribosomes.
✓ Actively transports Ca++ and stores it in the perinuclear space.
Nuclear envelope (1)
B. Nuclear Pore:
1. Definition:
Nuclear pores: are :
♦ Complex structures consisting of interrupted zones of the nuclear envelope,
4 Academic year: 2019-2020
Oran1 Ahmed Benbella University, Faculty of Medicine, Department of Medicine
Histology-Embryology Department, Dr Belarbi-Amar
N
Cytology class for first-year medical students
♦ Formed by an assembly of positively charged proteins (around fifty) called
nucleoporins.
♦ With a molecular weight of around 125 million Daltons, involved inexchanges
between the nucleus and 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 cell's physiological state, particularly
during cell growth (embryogenesis).
3. three-dimensional structure:
It is organized into 8 subunits, forming two rings, one cytoplasmic, the other
nucleoplasmic. Each ring carries perpendicular filaments, the filaments on the nucleoplasmic
side being joined 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 leave a central
tunnel of the order of 40nm. The central tunnel is closed by a diaphragm, opening when
molecules pass through in the presence of energy.
4. Role of the pore:
The 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
pore's lateral channels.
• The transport of large molecules occurs through the central transporter and sits 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 adapter (importine, exportine) recognizing an addressing
system.
5 Academic year: 2019-2020
Oran1 Ahmed Benbella University, Faculty of Medicine, Department of Medicine
Histology-Embryology Department, Dr Belarbi-Amar
N
Cytology class for first-year medical students
Cytoplasmic
filaments
Cytoplasmic ring
Nuclear ring
Nuclear basket
Cross-section of a nuclear pore
(2)
6 Academic year: 2019-2020
Oran1 Ahmed Benbella University, Faculty of Medicine, Department of Medicine
Histology-Embryology Department, Dr Belarbi-Amar
N
Cytology class for first-year medical students
The nuclear pore complex
Front view (4)
Transport through the pore (4)
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Oran1 Ahmed Benbella University, Faculty of Medicine, Department of Medicine
Histology-Embryology Department, Dr Belarbi-Amar
N
Cytology class for first-year medical students
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.
◾ Role: 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 encoding lamin A on
chromosome 1.
8 Academic year: 2019-2020
Oran1 Ahmed Benbella University, Faculty of Medicine, Department of Medicine
Histology-Embryology Department, Dr Belarbi-Amar
N
Cytology class for first-year medical students
E. Chromatin :
1. Definition: chromatin is the content of nucleoplasm observed by light or
electron microscopy in the nuclei of cells in interphase (outside mitosis).
Chromatin takes two forms:
• Heterochromatin: darkened 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 dense
under electron microscopy, representing 10 to 20% of adult cell chromatin
and corresponding to metabolically active regions (transcription).
2. Distribution:
a) Euchromatin is distributed throughout the nucleoplasm. b)
Heterochromatin, on the other hand, is found in the
nucleoplasm.
⮚ 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 cell activity. It takes the
form of :
♦ Small or large clods.
♦ Granulations or sprays.
♦ Confluent mesh networks in places.
Barr's corpuscle (sex chromatin): a mass of heterochromatin found in somatic
cell nuclei
9 Academic year: 2019-2020
Oran1 Ahmed Benbella University, Faculty of Medicine, Department of Medicine
Histology-Embryology Department, Dr Belarbi-Amar
N
Cytology class for first-year medical students
1um in diameter, corresponding to one of the X chromosomes (inactivated).
Electron microscopy: Chromatin ultra-structure 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 cell activity. A cell undergoing intense
activity has a high level of decondensed (clear) chromatin.
The nucleus under electron microscopy
10 Academic year: 2019-2020
Oran1 Ahmed Benbella University, Faculty of Medicine, Department of Medicine
Histology-Embryology Department, Dr Belarbi-Amar
N
Cytology class for first-year medical students
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) required 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.
11 Academic year: 2019-2020
Oran1 Ahmed Benbella University, Faculty of Medicine, Department of Medicine
Histology-Embryology Department, Dr Belarbi-Amar
N
Cytology class for first-year medical students
12 Academic year: 2019-2020
Oran1 Ahmed Benbella University, Faculty of Medicine, Department of Medicine
Histology-Embryology Department, Dr Belarbi-Amar
N
Cytology class for first-year medical students
DNA structure
13 Academic year: 2019-2020
Oran1 Ahmed Benbella University, Faculty of Medicine, Department of Medicine
Histology-Embryology Department, Dr Belarbi-Amar
N
Cytology class for first-year medical students
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 range 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: the nucleolus under light microscopy:
Appears as a spherical or oval refractive body, surrounded by a ring of chromatin.
It includes:
-Perinucleolar chromatin: in the form of a crescent more or less completely
surrounding the nucleolar body.
-The nucleolar body: appears as a homogeneous dense body, measuring 1
to 2um in normal cells.
3.Nucleolus components in ME: 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 fibrillar centers and dense granular
components are embedded.
4. Organization of the nucleolus:
The nucleolus contains large loops of DNA, belonging t o five pairs of acrocentric
chromosomes13, 14, 15, 21, 22, each of which contains a group of rRNA genes - around
forty per chromosome. Each of these groups constitutes a nucleolar organizing center or
NOR.
The three nucleolar regions:
1 The fibrillar center: containing proteins involved in transcription
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Oran1 Ahmed Benbella University, Faculty of Medicine, Department of Medicine
Histology-Embryology Department, Dr Belarbi-Amar
N
Cytology course for first-year medical students
such as RNA polymerase I and transcription factors.
2. Dense fibrillar component: contains a protein called fibrillarin, involved in rDNA
transcription and pre-rRNA cleavage.
3. Granular component: containing prebosomes.
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 RNA associates with imported ribosomal
proteins to form 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 5s
RNA (which is transcribed from extranuclear DNA by the
polymerase
15 Academic year: 2019-2020
Oran1 Ahmed Benbella University, Faculty of Medicine, Department of Medicine
Histology-Embryology Department, Dr Belarbi-Amar
N
Cytology course for first-year medical students
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 in the study of cell cycle
blockade following DNA strand damage caused by physical (e.g. ionizing
radiation) or chemical (e.g. pesticides) aggression.
6. Nucleoli and diseases:
◾ 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: 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).
16 Academic year: 2019-2020
Oran1 Ahmed Benbella University, Faculty of Medicine, Department of Medicine
Histology-Embryology Department, Dr Belarbi-Amar
N
Cytology course for first-year medical students
13, 14, 15, 21 and 22
17 Academic year: 2019-2020
Oran1 Ahmed Benbella University, Faculty of Medicine, Department of Medicine
Histology-Embryology Department, Dr Belarbi-Amar
N
Cytology course for first-year medical students
Core
Cytoplasm
18 Academic year: 2019-2020
Oran1 Ahmed Benbella University, Faculty of Medicine, Department of Medicine
Histology-Embryology Department, Dr Belarbi-Amar
N
Cytology course for first-year medical students
Morphological characteristics of cancer cells :
Morphologically, cancer cells may show numerous modifications of normal cell
characteristics, but n o cytological criteria are specific to cancer.
Intra-cellular organelles may be affected in size, shape or function.
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 cores.
• Increased nucleocytoplasmic ratio.
• Chromatin condensation, or nuclear hyperchromatism.
• Nucleolar abnormalities: enlarged, irregular or multinucleated nucleoli.
2- Mitosis abnormalities: the number and atypical nature of mitoses are indicative of
malignancy.
• Increase in the number of mitoses with shortening of the interphase.
• Abnormal, multipolar mitoses with irregular distribution of chromosomes.
3- Cytoplasmic abnormalities :
• Anisocytosis: cells of irregular size and shape, cellular gigantism.
• The cytoplasm is reduced and basophilic.
• Accumulation of certain elaboration products e.g. keratin, mucus, glycogen.
4- Plasma membrane abnormalities :
- light microscopy: it appears thickened and irregular.
- electron microscopy: it is bristling with amorphous, irregular microvilli, and
cell junctions are also affected.
19 Academic year: 2019-2020
Oran1 Ahmed Benbella University, Faculty of Medicine, Department of Medicine
Histology-Embryology Department, Dr Belarbi-Amar
N
Cytology course for first-year medical students
References :
1. Cell Biology. Abrégés. Marc Maillet.9th edition, Masson2002.
2. Cell Biology. Y Bassaglia. Maloine 2001.
3. Cell Biology .MC Dscamps. 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.
20 Academic year: 2019-2020
