We
have studied the hypothalamus of six male-to-female transsexuals (T1-T6);
this material that was collected over the last eleven years. We searched for
a brain structure that was sexually dimorphic, but not influenced by sexual
orientation, as male-to-female transsexuals may be "oriented" to
either sex with respect to sexual behavior. Our earlier observations showed
that the paraventricular nucleus (PVN),
sexually dimorphic nucleus (SDN) and suprachiasmatic nucleus (SCN) did not
meet these criteria ([6] and unpublished data). Although there is no accepted
animal model for gender identity alterations, the bed nucleus of the stria
terminalis (BST) turned out to be an appropriate candidate to study for the
following reasons. First, it is known that the BST plays an essential part in
rodent sexual behavior [3,4]. Not only have estrogen and androgen receptors
been found in the BST [8,9], it is also a major aromatization centre in the
developing rat brain [10]. The BST in the rat receives projections mainly
from the amygdala and provides a strong input in the preoptic-hypothalamic
region [11,12]. Reciprocal connections between hypothalamus, BST and amygdala
are also well documented in experimental animals [13-15]. In addition, sex
differences in the size and cell number of the BST have been described in
rodents which are influenced by gonadal steroids in development [16-18]. Also
in humans a particular caudal part of the BST (BNST-dspm) has been reported
to be 2.5 times larger in men than in women [19].
The
localization of the BST is shown in figure 1. The central part of the BST
(BSTc) is characterized by its somatostatin cells and vasoactive intestinal
polypeptide (VIP) innervation
[20]. We measured the volume of the BSTc on the basis of its VIP innervation (Fig. 2).
The
BSTc volume in heterosexual men (2.49±0.16 mm3) was 44% larger
than in heterosexual women (1.73±0.13 mm3) (P<0.005) (Fig. 3).
The volume of the BSTc of heterosexual and homosexual men was found not to
differ in any statistically significant way (2.81±0.20 mm3)
(P=0.26). The BSTc was 62% larger in homosexual men than in heterosexual
women (P<0.005). AIDS did not seem to influence the size of the BSTc: the
BSTc size of two heterosexual AIDS-infected women and three heterosexual
AIDS-infected men remained well within the range of the corresponding
reference group (Fig. 3). The AIDS-infected heterosexuals were therefore
included in the corresponding reference group for statistical purposes. A
small volume of the BSTc (1.30±0.23 mm3) was found in the
male-to-female transsexuals (Fig. 3). Its size was only 52% of that found in
the reference males (P<0.005) and 46% of the BSTc of homosexual males
(P<0.005). Although the mean BSTc volume in the transsexuals was even
smaller than that in the female group, the difference did not reach
statistical significance (P=0.13). The volume of the BSTc was not related to
age in any of the reference groups studied (P>0.15), indicating that the
observed small size of the BSTc in transsexuals was not due to the fact that
they were, on average, 10 to 13 years older than the hetero- and homosexual
men.
The
BST plays an essential role in masculine sexual behavior and in the
regulation of gonadotrophin release, as shown by studies in the rat [3,4,21].
There has been no direct evidence that the BST has such a role in human
sexual behavior but our demonstration of a sexually dimorphic pattern in the
size of the human BSTc, which is in agreement with the previously described
sex difference in a more caudal part of the BST (BNST-dspm) [19], indicates
that this nucleus may also be involved in human sexual or reproductive
functions. It has been proposed that neurochemical sex differences in the rat
BST may be due to effects of sex hormones on the brain during development and
in adulthood [22,23]. Our data from humans however, indicate that BSTc volume
is not affected by varying sex hormone levels in adulthood. The BSTc volume
of a 46-year-old woman who had suffered for at least 1 year from a tumour of
the adrenal cortex that produced very high blood levels of androstenedione
and testosterone, was within the range of that of other women (Fig. 3: S1).
Furthermore, two postmenopausal women (aged over 70 years) showed a completely
normal female-sized BSTc (Fig. 3: M1, M2). As all the transsexuals had been
treated with estrogens, the reduced size of the BSTc could possibly have been
due to the presence of high levels of estrogen in the blood. Evidence against
this comes from the fact that transsexual T2 and T3 both showed a small,
female-like BSTc (Fig. 3), although T2 stopped taking estrogen about 15
months before death, since her prolactin levels were too high and T3 stopped
hormone treatment since a sarcoma was found about three months before death;
also a 31-year-old man who suffered from a feminizing adrenal tumour which
induced high blood levels of estrogen, nevertheless had a very large BSTc
(Fig. 3: S2).
Our
results might also be explained if the female-sized BSTc in the transsexual
group was due to the lack of androgens, because they had all been
orchidectomized except for T4. We therefore studied two other men who had
been orchidectomized because of cancer of the prostate (one and three months
before death: S4 and S3, respectively), and found that their BSTc sizes were
at the high end of the normal male range. The BSTc size of the single
transsexual who had not been orchidectomized (T4) ranged in the middle of the
transsexual scores (Fig. 3). Not only were five of the transsexuals
orchidectomized, they all used the antiandrogen cyproterone acetate (CPA). A
CPA effect on the BSTc does not seem likely, because T6 had not taken CPA for
the past 10 years, and T3 took no CPA during the two years before death and
still had a female-sized BSTc.
In
summary, our observations suggest that the small size of the BSTc in
male-to-female transsexuals cannot be explained by differences in adult sex
hormone levels, but is established during development by an organizing action
of sex hormones, an idea supported by the fact that neonatal gonadectomy of
male rats and androgenization of the female rats indeed induced significant
changes in the number of neurons of the BST and suppressed its sexual
dimorphism [17,18].
Considered
together with information from animals, then our study supports the
hypothesis that gender identity alterations may develop as a result of an
altered interaction between the development of the brain and sex hormones
[5,6]. The direct action of genetic factors should also be considered on the
basis of animal experiments [24].
We
found no relationship between BSTc size and the sexual orientation of
transsexuals, that is, whether they were male-oriented (T1,T6),
female-oriented (T3,T2,T5), or both (T4). Furthermore, the size of the BSTc
of heterosexual men and homosexual men did not differ, which reinforced the
idea that the reduced BSTc size is independent of sexual orientation. In
addition, there was no difference in BSTc size between early-onset (T2,T5,T6)
and late-onset transsexuals (T1, T3), indicating that the decreased size is
related to the gender identity alteration per se rather than to the age at
which it becomes apparent. Interestingly, the very small BSTc in transsexuals
appears to be a very local brain difference. We failed to observe similar
changes in three other hypothalamic nuclei, namely, PVN,
SDN or SCN in the same individuals (unpublished data). This might be due to
the fact that these nuclei do not all develop at the same time, or to a
difference between these nuclei and the BST with respect to the presence of
sex hormone receptors or aromatase. We are now studying the distribution of
sex hormone receptors and the aromatase activity in various hypothalamic
nuclei in relation to sexual orientation and gender.
We
thank Mr. B. Fisser, Mr. H. Stoffels, Mr. G. van der Meulen, and Ms. T.
Eikelboom and Ms. W.T.P. Verweij for their help, and Drs. R.M. Buijs, M.A.
Corner, E. Fliers, A. Walter and F.W. van Leeuwen for their comments. Brain
material was provided by the Netherlands Brain Bank (coordinator Dr. R.
Ravid). This study was supported by NWO.
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Correspondence
and requests for materials to:
J.-N. Zhou, M.A. Hofman and D.F. Swaab
Graduate School Neurosciences Amsterdam
Netherlands Institute for Brain Research
Meibergdreef 33
1105 AZ Amsterdam ZO
The Netherlands
L.J.G.
Gooren
Department of Endocrinology
Free University Hospital
1007 MB Amsterdam
The Netherlands
Email: lgooren@inter.nl.net
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