BARR BODIES IN TESTIS WITH

KLINEFELTER SYNDROME

A. K. M. SHAMSUDDIN, M.D., PH.D.

CHIK-KWUN TANG, M.B.

From the Department of Pathology, University of Maryland School of Medicine, Baltimore, Maryland

ABSTRACT - Barr bodies are described in the interstitial cells of testes of a chromatin-positive patient with Klinefelter syndrome. Ultrastructural studies confirm the origin of the Barr body from the nuclear chromatin. Ultrastructurally the interstitial cells showed diminished, smooth endo- plasmic reticulum and lipid droplets probably indicating impaired function.

In the evaluation of patients with Klinefelter syndrome buccal mucosa is the usual tissue to look for Barr bodies. Although Ferguson-Smith et al.’ separated the histologic features of the testes in Klinefelter syndrome into chromatin- positive and chromatin-negative groups, testic- ular Barr bodies were not included in the description distinguishing one group from the other. We have recently encountered nuclear structures that are identical to Barr bodies in the interstitial cells of testes removed from a patient with Klinefelter syndrome and wish to report the histologic and ultrastructural findings.

Case Report

A thirty-four-year-old Caucasian man was diagnosed to have Klinefelter syndrome with chromosomal studies showing a XXY pattern. He had had retractile testes which caused severe pain when they were retracted and when he tried to lift heavy objects. The patient’s body height was greater than the arm span. He had a normal male hair pattern. The patient was on Depo-testosterone therapy. The testes were removed to relieve his pain.

Pathologic findings On gross examination the testes weighed 5.3

Gm. each. They were small and shrunken with thick capsule. Microscopically, the testes showed very thick tunica albuginea. Most of the seminiferous tubules were hyalinized. The re-

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maining tubules had thick basement mem- branes and were lined by Sertoli cells. Only a few seminiferous tubules showed some germ cells. The tubules were surrounded by fibrous tissue in which islands of interstitial cells (Ley- dig cells) were present (Fig. 1). These interstitial cells were pleomorphic but invariably contained eosinophilic cytoplasm. No definite crystals of Reinke were observed. Prominent nucleoli were seen. In some nuclei, triangular or hemi- oval basophilic bodies identical to Barr bodies were seen blending with the nuclear membranes.

Tissues from one of the two testes for electron microscopic examination were fixed adequately in formaldehyde (4%) and glutaraldehyde (1%) mixture at 176 mOsm., were washed with sucrose cacodylate buffer, osmicated, stained en bloc in uranyl acetate, dehydrated in graded ethanol, and embedded in Epon-812. One- micron thick sections were stained with tolui- dine blue and representative areas were selec- ted for thin sectioning. 600-800 A thin sections were stained with lead citrate and uranyl ace- tate and examined with a JEOL 100 B electron microscope at 60 Kv.

Electron microscopic study revealed large interstitial cells with prominent profiles of smooth endoplasmic reticulum some of which were dilated and many others were vesicular. There were scanty mitochondria, some with tubular cristae. Abundant Golgi profiles and stacks of rough endoplasmic reticulum were also

UROLOGY / JANUARY 1980 / VOLUME XV, NUMBER 1

FIGURE 1. Section oj testis showing hyalinized seminiferous tubules lined by Sertoli cells and groups of interstitial cells with pleomorphic bizarre nuclei (hematoxylin-eosin stain). Inset shows Barr body in paraffin- embedded hematoxylin- eosin section. (Original magnilfications, x 80, x 1,260, respectively.)

FIGURE 2. (A) Interstitial cell of ikydig showing microvillous projection of cell membrane, stacks of rough endoplasmic reticulum, scanty smooth endo- plasmic reticulum, no lipid, and tubular cristae of mitochondria. (B) Nucleus of interstitial cell showing trapezoid area of clumped chromatin (left side); chromatin material considered to be Barr body has morphology identical to rest of chromatin. (Original mugnifiations X 4,000 and X 8,000 respectively.)

UROLOGY / JANUARY 1980 / VOLUME XV, NUMBER 1

seen, The cc>11 membrane showed some micro- villous projections (Fig. 2A). Crystals of Reinke were not observed. Triangular clumps of elec- tron-dense granular structures corresponding to the Barr bodies seen on light microscopy were found along the nuclear membrane (Fig. 2B). The granular pattern of these structures was in- distinguishable from that of the adjacent chromatin and was distinctly different from that of the nucleoli which were prominent. Lipid droplets were not observed. Abundant finely granular and fibrillar moderately electron- dense substance was present in between organelles. Sertoli cells lining the tubules were elongated with relatively straight lateral cell membrane without folding. Desmosomes were conspicuous. There were scanty amounts of organelles, including a few stacks of rough endo- plasmic reticulum, irregular lipid inclusions, a few mitochondria, bundles of cytoplasmic fila- ments and crystalloid of Charcot-Bottcher; the basal lamina appeared to be reduplicated (Fig. 3).

Comment

As described by Ferguson-Smith et al., 1 the testes from chromatin-positive cases showed aggregates or masses of interstitial cells with pleomorphic nuclei. Large areas devoid of tubules were found. The remaining tubules were either totally hyalinized or lined by Sertoli cells. Spermatogenesis was rare. The testes from the chromatin-negative cases showed diffusely increased interstitial cells that were cytologically normal. The seminiferous tubules were of normal size, but most were lined solely by Sertoli cells. In all cases, a few tubules showed some spermatogenesis. The

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FIGURE 3. Seminiferous tubule showing redupli- cated basal lamina, tall elongated Sertoli cells, lipid vacuole, stacks of rough endoplasmic reticulum, straight lateral membrane, cytoplasmic filaments, and Charcot-Biittcher crystalloid (original magnijca- tion x 4,000). Inset: Higher magnijcation of Charcot-B6ttcher crystalloid.

light microscopic findings in our patient, there- fore, belong to the chromatin-positive category. Additionally, triangular or hemi-oval dark purple structures were found attached to the nuclear membrane of the nuclei of some inter- stitial cells. These structures demonstrated the same staining characteristics as that of the nucleoli found within the same nuclei, appear- ing dark purple on hematoxylin-eosin stain, and are interpreted as identical to Barr bodies found in the buccal mucosa.

The granular pattern found ultrastructurally in the area corresponding to Barr bodies on

light microscopic examination were indis- tinguishable from that of the adjacent chroma- tin. This finding, in conjunction with an extra number of sex chromosomes, indicates that Barr bodies are indeed sex chromosomes.

The function of interstitial cells is known to be impaired to varying degrees despite the fact that they are hyperplastic.2 In our patient, the impaired function of the interstitial cells is suggested by the ultrastructural findings when compared with those of interstitial cells in a man of normal androgenic status.3 Although profiles of smooth endoplasmic reticulum were prom- inent in the interstitial cells of the testis under study, they were not as packed as those in interstitial cells that actively produced andro- genie hormone. The reduction of smooth endo- plasmic reticulum, accompanied by the lack of lipid droplets which are conspicuous in active interstitial cells suggests that this patient’s interstitial cells probably were not as active as normal interstitial cells. However, due to lack of correlative laboratory data, no valid con- clusion can be drawn on morphologic changes alone. Whether or not the presence of the finely fibrillar material in between the organelles reflects an impaired function of the interstitial cells is not known. As in normal Sertoli cells, the Sertoli cells in this testis contained lipid droplets in cytoplasm, bundles of cytoplasmic filaments, desmosomes, and Charcot-Biittcher crystalloid.4 Different from the normal Sertoli cells, however, were scanty, smooth endo- plasmic reticulum and stacks of rough endo- plasmic reticulum. Lamellar bodies which are also features of normal Sertoli cells were not found in this testis. Whether or not this mor- phologic difference reflects functional dis- turbance cannot be evaluated since the function of the Sertoli cell is not clear.

22 South Green Street Baltimore, Maryland 21201

(DR. SHAMSUDDIN)

References

1. Ferguson-Smith MA, Lennox B, Mack WS, and Stewart JSS: Klinefelter’s syndrome. Frequency and testicular morphology in relation to nuclear sex. Lancet 2: 167 (1957).

2. Wang T-W, Straus FH, and Warner NE: Testicular biopsy in the study of male infertility. I. Testicular causes of infertility, Arch. Pathol. 95: 151 (1973).

3. DeKrester DM: The fine structure of the testicular interstitial cells in men of normal androgenic status, Z. Zellforsch. Mikrosk. Anat. 80: 594 (1967).

4. Nagano T: Some observations on the fine structure of the Sertoli cell in the human testis, ibid. 73: 89 (1966).

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