University of Bahrain
College of Science
Department of Biology
Molecular and Cellular Characterization of Normal Versus Aberrant Sperm from Fertility Test
Groups
Sayed Hashim Ebrahim, Supervised by Dr.Maheen Dairi and Dr. Abdulla A. Rasool
University of Bahrain, College of Science, Sakhir.
Abstract
Human semen samples from individuals subjected
to fertility test at the infertility clinic of Salmaniya
Medical Complex were characterized by applying
molecular and cellular analyses of semen
samples. Data analysis covered semen volume,
sperm count, liquefaction time, semen pH, sperm
motility and deformation following the WHO 1992
criteria.
The results showed that 79% of individuals had
normal sperm count, 74% had normal pH and 85%
had normal volume. However, liquefaction time
was aberrant in 65% of individuals. Further
analysis on normal sperm count subgroups and
showed that 26% had abnormal pH, 62% had
abnormal liquefaction time and 16% had abnormal
volume. The results were further supported by
calculating the statistical correlation between the
different parameters. For example negative
correlations (p=0.01) was found between
liquefaction and two parameters: volume and
sperm count. On the other hand, positive
correlations were found between age and volume\
sperm count. Assuming that other molecular
factors are normal, only 18% of the individuals
were potentially fertile.
Aim
nucleoprotamine) During spermiogenesis, nuclear
re-modeling and condensation is associated with
the sequential displacement of histones by
transition proteins and then by protamines (PRM1
and PRM2) (2).
The aim of the project was to characterize human semen
samples from individuals subjected to fertility test at the
infertility clinic of Salmaniya Medical Complex by
applying molecular and cellular analyses of semen
samples.
Materials & Methods
Sperm
Collection
Semen
Analysis
Sperm
Staining
Morphometric
Analysis
Loading &
Viewing the
Stained DNA
Agarose Gel
Preparation for
Electrophoresis
Chromatin
Extraction &
Digestion
Sperm
Preparation &
Fixation
Results
Figure 3: Percentage of individuals with Normal pH = 7-8;
Abnormal pH > 8.
Figure 1: Percentage of sperm count. Of Normal (20-50
million sperm/ml) and Abnormal (<20 million sperm/ml).
Figure 2: Percentage of sperm count of Oligospermic
(< 20 million sperm/ml) & Azoospermic (no sperm).
Figure 5: Relationship between sperm count
subgroups and liquefaction time.
Figure 6: Percentage analysis of individuals with normal
(1.5-5ml) & abnormal seminal fluid volume (<1.5 ml).
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Figure 7: Relationship between sperm count subgroups &
seminal fluid volume.
Figure 18: A prepared
slide for human sperm
with coiled tail.
Figure 4: Relationship between sperm count subgroups and
liquefaction time.
Figure 8: Analysis of sperm deformation in total samples.
Deformation is the % measurement of defects (< 60% normal
sperm) in tail, head, & neck.
Figure 22: Gel electrophoresis photo represents the
digestion reaction of samples 3 & 6 using DNaseI
partial digestion. Lanes 1–3 and lanes 4–6 represents
sperm samples 3 & 6, respectively. The two samples
were treated with DNaseI partial digestion as shown in
the table below. Top arrow show intact DNA whereas
the bottom arrow points at a band generated after
digestion with DNaseI. note that the migration of all
bands are similar.
Figure 20: A prepared
slide for human sperm
with double tail.
Discussion & Conclusion
Average length of head, neck and tail of normal
sperm were 4.5, 7 and 45 um respectively. The
average width of normal sperm head was 2.5 um.
These data are consistent with those reported by
WHO (1992) and the work done on orientation of
sea urchin and human sperm in static magnetic
field (3).
A significant positive correlation between age
and most of the semen parameters was recorded.
Also 18 % of the samples showed a potential
healthy condition for all semen parameters.
Assuming that all other molecular factors like,
gene structure, or other biochemical factors are
normal, then these individuals were referred to
the clinic for double check up and the fertility
test can be further focused on their wives.
The majority of samples showed abnormal
semen liquefaction. This problem could be
medicinally treatable especially that 80% of the
samples have normal sperm count.
References
1Watanabe
Figure 9: Analysis of liquefaction time in
individuals with the normal sperm count.
Figure 10: Analysis of seminal fluid pH in individuals
with the normal sperm count.
Figure 11: Relationship between sperm count subgroups
and pH
Figure 12: Relationship between
sperm count subgroups and volume
Light Microscopic Examination (1000X )
T.,et al. (2007). J.Congenital Anomalies. 47(1): 9-15.
2Esmaeilzadeh S., et al. (2007). J. Reprod.& infertility. 22( 11): 133-140.
3Sakhnini L & Dairi M. (2007). J. Magnetism and Magnetic Materials. 310(2): 1035-1037.
4Balakrishnan, L., & Milavetz B., (2007). J. Mole. Biol. 371(4): 1022-1037.
5Levitas E., et al. (2007). J. Andrlogia. 39(2):45-50
Acknowledgment
Figure 13: A prepared slide
for human sperm with an
amorphous head
POSTER TEMPLATE BY:
Figure 21: A prepared
slide for human
sperm with kinked
tail.
Figure 19: A prepared slide
for human sperm with short
tail.
Introduction
Infertility is defined as not being able to get
pregnant after at least one year of trying. In women,
irregular or absent menstrual periods may indicate
that the female is not capable of ovulation.
The evaluation of a subfertile man includes a
comprehensive history, physical examination,
multiple semen analyses and an endocrine
evaluation.
Environmental factors such as temperature,
Smoking, Dietary deficiencies such as vitamins A,
B, and E, x-ray exposure, alcohol and infectious
diseases adversely affect the rate of
spermatogenesis (1). The DNA in human sperm
chromatin is bound to nucleohistone and
Figure 20: A prepared slide for
human sperm with absence mid
piece region.
Figure 14: A prepared slide for
abnormal human sperm with
very small size of head
(Microsperm).
Figure 15: A prepared
slide for human sperm
with an elongated head.
Figure 16: Human sperm
with a tapered head.
Figure 17: A prepared
slide for human sperm
with double head
abnormality.
Figure 18: A prepared
slide for human sperm
with rounded head.
Figure 19: A prepared slide
for human sperm with
Kinked mid piece region.
I would like to thank Dr. Maheen Dairi and Dr. Abdulla A.Rasool for their supervision, the technical staff at
UOB for laboratory assistance and Salmanya Hospital for providing samples from patients.