CORRELATION BETWEEN THE CONCENTRATION

OF \g=b\-TRACE PROTEIN AND THE NUMBER OF

SPERMATOZOA IN HUMAN SEMEN
J. E. OLSSON
Departments of Neurology and Forensic Medicine,
University Hospital, S-221 85 Lund, Sweden
(Received 17th June 1974)
Beta-trace protein (\g=b\-TP) is predominantly associated with the central nervous
system (CNS) and was first discovered in concentrated cerebrospinal fluid
(CSF) by Clausen (1961). The protein has a low molecular weight of 31,000
and is present in high concentration in the CSF (0\m=.\8to 3\m=.\9mg/100 ml), con-
stituting about 7% of the total CSF protein concentration (Link, 1967). By
contrast, the serum concentration of the protein is low (0\m=.\26to 0\m=.\60mg/100 ml)
and \g=b\-TP constitutes only 1/20,000 of the total serum protein content (Olsson,
Link & Nosslin, 1973). The protein is also found in urine (Hochwald & Thor-
becke, 1962), in which the daily excretion is below 9\m=.\5mg (Ericsson, Link &
Zettervall, 1969), as well as in various organ extracts, e.g. brain, kidney,
pancreas and genital organs (Laterre, Heremans & Carbonara, 1964; Penny &
Osserman, 1971). The highest amounts of \g=b\-TP are found in white matter
(Olsson & Link, 1973) and in the epididymis (Olsson & Nord, 1973). In the
former, \g=b\-TP is mostly found in the glial cells and in brain tumours of glial
cell origin (Olsson, Blomstrand & Haglid, 1974).
An increased concentration of /?-TP is found in the CSF after strokes involving
brain damage (Link & Olsson, 1972) and in severely disabled patients with
multiple sclerosis (J. E. Olsson, H. Link and R. Müller, unpublished observa¬
tions), and its excretion in urine is increased in patients with brain tumours,
cerebrovascular diseases and renal failure (Ericsson et al., 1969). Recently,
another clinical application of /?-TP, as a marker of fetal neural-tube defects
in samples of amniotic fluid, has been reported (Macri, Weiss, Joshi & Evans,
1974).
Interest in the curious anatomical distribution of /?-TP, especially in the
CNS and the genital organs and its high concentration in the CSF, initiated
the present study of the protein in normal and pathological human seminal
fluid.
Samples of semen were obtained from 309 men, who were examined because
of barren marriages or as a link in a paternity examination. The numbers of
spermatozoa were counted and the patients were allocated to four groups
according to the number of cells. In addition, the motility and the morphology
of the cells were examined using a light microscope. The different groups of
patients are shown in Table 1. In all samples, the concentration of jS-TP was
determined by single radial immunodiffusion on agar as described by Mancini,
149

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150 J. E. Olsson
Carbonara & Heremans (1965) and the total protein content was assessed by
the method described by Lo wry, Rosebrough, Farr & Randall (1951).
Semen samples were obtained from eight men whose ejaculates showed
'normal' sperm morphology, motility and concentration. The samples were
centrifuged three times at 9000 g for 15 min on each occasion. The resulting
supernatant was free from spermatozoa when examined using a light micro¬
scope. The total protein content and the concentration of /?-TP were deter¬
mined in the original sample, the pellet and the supernatant.
A specific antiserum against /?-TP was prepared by repeated immunization
of rabbits with pure /?-TP obtained from human CSF (Link, 1967). The

Table 1. Number of spermatozoa, concentration of ß-trace

protein and total protein content in human semen in different
groups of patients

No. of sperm. ß-trace protein Total protein

( 10* ¡ml) (mg/100 ml) [gl 100 ml)
40 to 250 (N =
140) 4-9 + 0-296 5-1 + 0-196
(0-6 to 180) (3-2 to 9-9)
10 to 40 (N =
89) 3-3 + 0-354 5-0±0-110
(0-3 to 21-6) (2-5 to 7-8)
< 10 (N =
61) 1-7 ±0-209 5-1 ±0-160
(0-0 to 8-9) (1-8 to 7-6)
Azoospermia (N =
19) 0-8 + 0-175 4-4 ±0-232
(0-0 to 3-5) (3-3 to 6-6)
Values are given with mean ± S.E.M. and range in parentheses.
= number of
patients.

antiserum was absorbed with human serum. An immunological identity

between human CSF and semen was found when testing these fluids against
antiserum against human /?-TP by the double diffusion technique described
by Ouchterlony (1948).
The concentration of /?-TP and the total protein concentration in semen
samples containing different numbers of spermatozoa are given in Table 1.
Comparisons between the mean numbers were performed with Student's t test.
A strong correlation was found between the concentration of /?-TP in human
seminal fluid and the number of sperm cells. The motility and the morphology
of the cells, however, did not appear to correlate with the content of /?-TP.
The concentration of /?-TP decreased when the number of spermatozoa
decreased and the values differed significantly ( <0·001) between all groups.
The total protein content was constant in all patients who had spermatozoa in
the semen, but was somewhat lower (P<0-05) in the azoospermic patients.
In the centrifuged samples of 'normal' semen, the concentration of /?-TP and
the total protein content remained fairly constant in the original sample and in
the supernatant of the centrifuged sample, whereas the concentration of /?-TP
was about 5 times lower and the total protein content about 2-5 times lower
in the pellet.

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 Beta-trace protein in human semen 151
In someof the samples, the content of fructose was determined. There was
no significant correlation between the concentrations of fructose and those of
jff-TP.
The present findings of a mean concentration of about 5 mg /?-TP/100 ml in
normal human semen indicate that the protein constitutes about 1/1000 of the
total protein concentration in semen. Thus, the relative concentration of
/?-TP in human semen is lower than that in CSF but higher than that in serum.
In patients with azoospermia, the mean relative concentration of /?-TP in
semen is only about 2/10,000 of the total protein content.
The connection between /?-TP and the number of spermatozoa in human
semen will be further investigated.

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