Aim: Analysis of the Ultrastructure of Mitochondria through Electron Micrographs.
Requirements:
● Electron micrographs of mitochondria
● Worksheet or drawing materials for labelling structures
● Reference diagrams from textbooks
Theory and Observations:
Introduction:
Mitochondria are organelles present in the cytoplasm of all eukaryotic cells. They provide an
energy-transducing system by which the chemical energy contained in foodstuffs is converted, by
oxidative phosphorylation, into high-energy phosphate bonds (ATP). Mitochondria may be observed in the
living cell; visibility is increased by the vital stain, Janus green. They display passive and active motion
and show changes in volume and shape that are related to their function.
History:
● Mitochondria were first observed in the 1850s by Rudolf Kölliker in muscle cells.
● Discovered by Richard Altmann in 1890, who named them "bioblasts".
● The term “mitochondria” was coined by Carl Benda in 1898, derived from Greek: mitos (thread)
and chondrion (granule).
● Walberg(1913) observed that respiratory enzymes were associated with cytoplasmic particles.
● The functional significance of mitochondria was understood after the discovery of oxidative
phosphorylation in the mid-20th century.
● A most important advance was the first isolation of liver mitochondria by Bensley and Hoerr in
1934. This established the possibility of a direct study of the organelles by biochemical methods.
● The final demonstration that the mitochondrion was indeed the site of cellular respiration was made
in 1948 by Hogeboom and co-workers.
Size and Dimensions:
● Mitochondria are double-membraned organelles.
● Dimensions: Typically 0.5–1.0 μm in diameter and 1.0–10 μm in length.
● Their number and size vary depending on the energy demands of the cell.
● Shape varies—commonly rod-shaped, but may be spherical, oval, or filamentous.
Ultrastructure:
Mitochondria are composed of the following components:
1. Outer Membrane:
○ Smooth, permeable to small molecules and ions.
○ Contains porins (proteins) that form channels.
2. Intermembrane Space:
○ Space between outer and inner membranes.
○ Important in oxidative phosphorylation.
3. Inner Membrane:
○ Impermeable; selectively allows passage through specific transporters.
○ Contains the electron transport chain and ATP synthase.
○ Folded into structures called cristae to increase surface area.
4. Cristae:
○ Extensions of the inner membrane.
○ Provide surface for oxidative phosphorylation enzymes.
5. Matrix:
○ Enclosed by the inner membrane.
○ Contains enzymes for the Krebs cycle, mitochondrial DNA, and ribosomes.
6. Mitochondrial DNA (mtDNA):
○ Circular, double-stranded.
○ Encodes some of the organelle's own proteins and RNAs.
7. Mitochondrial Ribosomes
○ 55–60S type (smaller than cytoplasmic ribosomes).
○ Involved in protein synthesis within the organelle.
Mitochondrial morphology and structure:
A mitochondria consists of two membranes and two compartments, An outer limiting membrane, about 6
nm thick, surrounds the mitochondrion. Within this membrane, and separated from it by a space of about 6
to 8 nm, is an inner membrane that projects into the mitochondrial cavity complex infoldings called
mitochondrial crests. This inner membrane divides the mitochondrion into two chambers or compartments:
(1) the outer chamber contained between the two mem-branes and in the core of the crests and
(2) the inner chamber is filled with a relatively dense proteinaceous material usually called the
mitochon-drial matrix. This is generally homogeneous, but in some cases it may contain a finely
filamentous material, or small, highly dense granules.
These granules contain phospholipids, which give them an affinity for calcium when this is added to the
fixative. There is no evidence, how-ever, for the normal presence of calcium phos-phate in the granules.
The mitochondrial crests are, in general, in-complete septa or ridges that do not interrupt the continuity of
the inner chamber, thus, the matrix is continuous within the mitochondrion.
Within the mitochondrial matrix are small ribosomes and a circular DNA. The outer and inner membranes
and the crests can be considered to be fluid molecular films with a compact molecular structure; the matrix
is gel-like and contains a high concentration of soluble proteins and smaller molecules. This double
(solid-liquid) structure is important in providing an explanation for some of the mechanical prop-erties of
mitochondria (e.g., deformation and swelling under physiological or experimental conditions).
F1 Particles On the M Side of the Inner Mitochondrial Membrane
The use of negative staining has enabled recognition of other details of mitochondrial structure. If a
mitochondrion is allowed to swell and break in a hypotonic solution and is then immersed in
phosphotungstate, the inner membrane in the crest appears covered by particles of 8.5 nm that have a
stem linking each with the membrane. These so-called "elementary" or "F1" particles are regularly spaced
at intervals of 10 nm on the inner surface of these membranes. According to some estimates, there are
between 10 and 10 elementary particles per mitochondrion. These particles correspond to a special
ATPase involved in the coupling of oxidation and phosphorylation.
Mitochondrial Structural Variations in Different Cell Types
A common mitochondrial structural pattern likely evolved early and persisted across organisms, from
protozoa to mammals and algae to flowering plants. However, variations exist: cristae may be longitudinal
(e.g., in nerve and muscle cells), simple, branched, or tubular (as in protozoa, insects, and adrenal cells).
The number of cristae varies with function—liver and germ cells have few cristae with abundant matrices,
while muscle cells, especially insect flight muscles, have many, sometimes densely packed in
quasi-crystalline arrays. Generally, more cristae correlates with higher oxidative activity.
There are 1000 to 1600 mitochondria in a liver cell and 300,000 in some oocytes. Green plants contain
fewer mitochondria than animal cells. The distribution of mitochondria may be related to their function as
suppliers of energy. Their orientation in the cell may be influ-enced by the organization of the cytoplasmic
matrix and vacuolar system.
Relationship to Lipids:
Since Altmann’s time, studies have noted a link between lipid distribution and mitochondrial activity. In
liver and pancreas cells, short-term starvation brings mitochondria into close contact with lipid droplets,
sometimes showing only the inner membrane adjacent to the lipid. This suggests active fat utilization via
mitochondrial fatty acid oxidases.
Intramitochondrial Inclusions:
Mitochondria can accumulate various substances—pigments from hemoglobin in amphibians, ferritin in
Cooley’s anemia, and crystalline protein masses when transformed into yolk bodies in mollusk eggs or
amphibian oocytes. These inclusions may appear in a regular crystalline arrangement.
Result:
The ultrastructure of mitochondria was successfully observed through electron micrographs, and its
structural components were identified and correlated with their respective functions.
Conclusion:
Mitochondria are dynamic, double-membraned organelles specialized for aerobic energy metabolism.
Their structure—particularly the inner membrane and cristae—is closely linked to their function of ATP
generation.
References:
● Cell and Molecular Biology by E.D.P. De Robertis
● Voet and voet
● Lehninger
● Cell Biology by Ambrose and Easty
● Cell and organelles by Holtzman and Novikoff
● Cell biology by Powar
